113 results on '"Keating, DJ"'
Search Results
2. Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)
- Author
-
Klionsky, DJ, Abdel-Aziz, AK, Abdelfatah, S, Abdellatif, M, Abdoli, A, Abel, S, Abeliovich, H, Abildgaard, MH, Abudu, YP, Acevedo-Arozena, A, Adamopoulos, IE, Adeli, K, Adolph, TE, Adornetto, A, Aflaki, E, Agam, G, Agarwal, A, Aggarwal, BB, Agnello, M, Agostinis, P, Agrewala, JN, Agrotis, A, Aguilar, PV, Ahmad, ST, Ahmed, ZM, Ahumada-Castro, U, Aits, S, Aizawa, S, Akkoc, Y, Akoumianaki, T, Akpinar, HA, Al-Abd, AM, Al-Akra, L, Al-Gharaibeh, A, Alaoui-Jamali, MA, Alberti, S, Alcocer-Gómez, E, Alessandri, C, Ali, M, Alim Al-Bari, MA, Aliwaini, S, Alizadeh, J, Almacellas, E, Almasan, A, Alonso, A, Alonso, GD, Altan-Bonnet, N, Altieri, DC, Álvarez, ÉMC, Alves, S, Alves da Costa, C, Alzaharna, MM, Amadio, M, Amantini, C, Amaral, C, Ambrosio, S, Amer, AO, Ammanathan, V, An, Z, Andersen, SU, Andrabi, SA, Andrade-Silva, M, Andres, AM, Angelini, S, Ann, D, Anozie, UC, Ansari, MY, Antas, P, Antebi, A, Antón, Z, Anwar, T, Apetoh, L, Apostolova, N, Araki, T, Araki, Y, Arasaki, K, Araújo, WL, Araya, J, Arden, C, Arévalo, M-A, Arguelles, S, Arias, E, Arikkath, J, Arimoto, H, Ariosa, AR, Armstrong-James, D, Arnauné-Pelloquin, L, Aroca, A, Arroyo, DS, Arsov, I, Artero, R, Asaro, DML, Aschner, M, Ashrafizadeh, M, Ashur-Fabian, O, Atanasov, AG, Au, AK, Auberger, P, Auner, HW, Aurelian, L, Autelli, R, Avagliano, L, Ávalos, Y, Aveic, S, Aveleira, CA, Avin-Wittenberg, T, Aydin, Y, Ayton, S, Ayyadevara, S, Azzopardi, M, Baba, M, Backer, JM, Backues, SK, Bae, D-H, Bae, O-N, Bae, SH, Baehrecke, EH, Baek, A, Baek, S-H, Baek, SH, Bagetta, G, Bagniewska-Zadworna, A, Bai, H, Bai, J, Bai, X, Bai, Y, Bairagi, N, Baksi, S, Balbi, T, Baldari, CT, Balduini, W, Ballabio, A, Ballester, M, Balazadeh, S, Balzan, R, Bandopadhyay, R, Banerjee, S, Bánréti, Á, Bao, Y, Baptista, MS, Baracca, A, Barbati, C, Bargiela, A, Barilà, D, Barlow, PG, Barmada, SJ, Barreiro, E, Barreto, GE, Bartek, J, Bartel, B, Bartolome, A, Barve, GR, Basagoudanavar, SH, Bassham, DC, Bast, RC, Basu, A, Batoko, H, Batten, I, Baulieu, EE, Baumgarner, BL, Bayry, J, Beale, R, Beau, I, Beaumatin, F, Bechara, LRG, Beck, GR, Beers, MF, Begun, J, Behrends, C, Behrens, GMN, Bei, R, Bejarano, E, Bel, S, Behl, C, Belaid, A, Belgareh-Touzé, N, Bellarosa, C, Belleudi, F, Belló Pérez, M, Bello-Morales, R, Beltran, JSDO, Beltran, S, Benbrook, DM, Bendorius, M, Benitez, BA, Benito-Cuesta, I, Bensalem, J, Berchtold, MW, Berezowska, S, Bergamaschi, D, Bergami, M, Bergmann, A, Berliocchi, L, Berlioz-Torrent, C, Bernard, A, Berthoux, L, Besirli, CG, Besteiro, S, Betin, VM, Beyaert, R, Bezbradica, JS, Bhaskar, K, Bhatia-Kissova, I, Bhattacharya, R, Bhattacharya, S, Bhattacharyya, S, Bhuiyan, MS, Bhutia, SK, Bi, L, Bi, X, Biden, TJ, Bijian, K, Billes, VA, Binart, N, Bincoletto, C, Birgisdottir, AB, Bjorkoy, G, Blanco, G, Blas-Garcia, A, Blasiak, J, Blomgran, R, Blomgren, K, Blum, JS, Boada-Romero, E, Boban, M, Boesze-Battaglia, K, Boeuf, P, Boland, B, Bomont, P, Bonaldo, P, Bonam, SR, Bonfili, L, Bonifacino, JS, Boone, BA, Bootman, MD, Bordi, M, Borner, C, Bornhauser, BC, Borthakur, G, Bosch, J, Bose, S, Botana, LM, Botas, J, Boulanger, CM, Boulton, ME, Bourdenx, M, Bourgeois, B, Bourke, NM, Bousquet, G, Boya, P, Bozhkov, PV, Bozi, LHM, Bozkurt, TO, Brackney, DE, Brandts, CH, Braun, RJ, Braus, GH, Bravo-Sagua, R, Bravo-San Pedro, JM, Brest, P, Bringer, M-A, Briones-Herrera, A, Broaddus, VC, Brodersen, P, Brodsky, JL, Brody, SL, Bronson, PG, Bronstein, JM, Brown, CN, Brown, RE, Brum, PC, Brumell, JH, Brunetti-Pierri, N, Bruno, D, Bryson-Richardson, RJ, Bucci, C, Buchrieser, C, Bueno, M, Buitrago-Molina, LE, Buraschi, S, Buch, S, Buchan, JR, Buckingham, EM, Budak, H, Budini, M, Bultynck, G, Burada, F, Burgoyne, JR, Burón, MI, Bustos, V, Büttner, S, Butturini, E, Byrd, A, Cabas, I, Cabrera-Benitez, S, Cadwell, K, Cai, J, Cai, L, Cai, Q, Cairó, M, Calbet, JA, Caldwell, GA, Caldwell, KA, Call, JA, Calvani, R, Calvo, AC, Calvo-Rubio Barrera, M, Camara, NO, Camonis, JH, Camougrand, N, Campanella, M, Campbell, EM, Campbell-Valois, F-X, Campello, S, Campesi, I, Campos, JC, Camuzard, O, Cancino, J, Candido de Almeida, D, Canesi, L, Caniggia, I, Canonico, B, Cantí, C, Cao, B, Caraglia, M, Caramés, B, Carchman, EH, Cardenal-Muñoz, E, Cardenas, C, Cardenas, L, Cardoso, SM, Carew, JS, Carle, GF, Carleton, G, Carloni, S, Carmona-Gutierrez, D, Carneiro, LA, Carnevali, O, Carosi, JM, Carra, S, Carrier, A, Carrier, L, Carroll, B, Carter, AB, Carvalho, AN, Casanova, M, Casas, C, Casas, J, Cassioli, C, Castillo, EF, Castillo, K, Castillo-Lluva, S, Castoldi, F, Castori, M, Castro, AF, Castro-Caldas, M, Castro-Hernandez, J, Castro-Obregon, S, Catz, SD, Cavadas, C, Cavaliere, F, Cavallini, G, Cavinato, M, Cayuela, ML, Cebollada Rica, P, Cecarini, V, Cecconi, F, Cechowska-Pasko, M, Cenci, S, Ceperuelo-Mallafré, V, Cerqueira, JJ, Cerutti, JM, Cervia, D, Cetintas, VB, Cetrullo, S, Chae, H-J, Chagin, AS, Chai, C-Y, Chakrabarti, G, Chakrabarti, O, Chakraborty, T, Chami, M, Chamilos, G, Chan, DW, Chan, EYW, Chan, ED, Chan, HYE, Chan, HH, Chan, H, Chan, MTV, Chan, YS, Chandra, PK, Chang, C-P, Chang, C, Chang, H-C, Chang, K, Chao, J, Chapman, T, Charlet-Berguerand, N, Chatterjee, S, Chaube, SK, Chaudhary, A, Chauhan, S, Chaum, E, Checler, F, Cheetham, ME, Chen, C-S, Chen, G-C, Chen, J-F, Chen, LL, Chen, L, Chen, M, Chen, M-K, Chen, N, Chen, Q, Chen, R-H, Chen, S, Chen, W, Chen, X-M, Chen, X-W, Chen, X, Chen, Y, Chen, Y-G, Chen, Y-J, Chen, Y-Q, Chen, ZS, Chen, Z, Chen, Z-H, Chen, ZJ, Cheng, H, Cheng, J, Cheng, S-Y, Cheng, W, Cheng, X, Cheng, X-T, Cheng, Y, Cheng, Z, Cheong, H, Cheong, JK, Chernyak, BV, Cherry, S, Cheung, CFR, Cheung, CHA, Cheung, K-H, Chevet, E, Chi, RJ, Chiang, AKS, Chiaradonna, F, Chiarelli, R, Chiariello, M, Chica, N, Chiocca, S, Chiong, M, Chiou, S-H, Chiramel, AI, Chiurchiù, V, Cho, D-H, Choe, S-K, Choi, AMK, Choi, ME, Choudhury, KR, Chow, NS, Chu, CT, Chua, JP, Chua, JJE, Chung, H, Chung, KP, Chung, S, Chung, S-H, Chung, Y-L, Cianfanelli, V, Ciechomska, IA, Cifuentes, M, Cinque, L, Cirak, S, Cirone, M, Clague, MJ, Clarke, R, Clementi, E, Coccia, EM, Codogno, P, Cohen, E, Cohen, MM, Colasanti, T, Colasuonno, F, Colbert, RA, Colell, A, Čolić, M, Coll, NS, Collins, MO, Colombo, MI, Colón-Ramos, DA, Combaret, L, Comincini, S, Cominetti, MR, Consiglio, A, Conte, A, Conti, F, Contu, VR, Cookson, MR, Coombs, KM, Coppens, I, Corasaniti, MT, Corkery, DP, Cordes, N, Cortese, K, Costa, MDC, Costantino, S, Costelli, P, Coto-Montes, A, Crack, PJ, Crespo, JL, Criollo, A, Crippa, V, Cristofani, R, Csizmadia, T, Cuadrado, A, Cui, B, Cui, J, Cui, Y, Culetto, E, Cumino, AC, Cybulsky, AV, Czaja, MJ, Czuczwar, SJ, D'Adamo, S, D'Amelio, M, D'Arcangelo, D, D'Lugos, AC, D'Orazi, G, da Silva, JA, Dafsari, HS, Dagda, RK, Dagdas, Y, Daglia, M, Dai, X, Dai, Y, Dal Col, J, Dalhaimer, P, Dalla Valle, L, Dallenga, T, Dalmasso, G, Damme, M, Dando, I, Dantuma, NP, Darling, AL, Das, H, Dasarathy, S, Dasari, SK, Dash, S, Daumke, O, Dauphinee, AN, Davies, JS, Dávila, VA, Davis, RJ, Davis, T, Dayalan Naidu, S, De Amicis, F, De Bosscher, K, De Felice, F, De Franceschi, L, De Leonibus, C, de Mattos Barbosa, MG, De Meyer, GRY, De Milito, A, De Nunzio, C, De Palma, C, De Santi, M, De Virgilio, C, De Zio, D, Debnath, J, DeBosch, BJ, Decuypere, J-P, Deehan, MA, Deflorian, G, DeGregori, J, Dehay, B, Del Rio, G, Delaney, JR, Delbridge, LMD, Delorme-Axford, E, Delpino, MV, Demarchi, F, Dembitz, V, Demers, ND, Deng, H, Deng, Z, Dengjel, J, Dent, P, Denton, D, DePamphilis, ML, Der, CJ, Deretic, V, Descoteaux, A, Devis, L, Devkota, S, Devuyst, O, Dewson, G, Dharmasivam, M, Dhiman, R, di Bernardo, D, Di Cristina, M, Di Domenico, F, Di Fazio, P, Di Fonzo, A, Di Guardo, G, Di Guglielmo, GM, Di Leo, L, Di Malta, C, Di Nardo, A, Di Rienzo, M, Di Sano, F, Diallinas, G, Diao, J, Diaz-Araya, G, Díaz-Laviada, I, Dickinson, JM, Diederich, M, Dieudé, M, Dikic, I, Ding, S, Ding, W-X, Dini, L, Dinić, J, Dinic, M, Dinkova-Kostova, AT, Dionne, MS, Distler, JHW, Diwan, A, Dixon, IMC, Djavaheri-Mergny, M, Dobrinski, I, Dobrovinskaya, O, Dobrowolski, R, Dobson, RCJ, Đokić, J, Dokmeci Emre, S, Donadelli, M, Dong, B, Dong, X, Dong, Z, Dorn Ii, GW, Dotsch, V, Dou, H, Dou, J, Dowaidar, M, Dridi, S, Drucker, L, Du, A, Du, C, Du, G, Du, H-N, Du, L-L, du Toit, A, Duan, S-B, Duan, X, Duarte, SP, Dubrovska, A, Dunlop, EA, Dupont, N, Durán, RV, Dwarakanath, BS, Dyshlovoy, SA, Ebrahimi-Fakhari, D, Eckhart, L, Edelstein, CL, Efferth, T, Eftekharpour, E, Eichinger, L, Eid, N, Eisenberg, T, Eissa, NT, Eissa, S, Ejarque, M, El Andaloussi, A, El-Hage, N, El-Naggar, S, Eleuteri, AM, El-Shafey, ES, Elgendy, M, Eliopoulos, AG, Elizalde, MM, Elks, PM, Elsasser, H-P, Elsherbiny, ES, Emerling, BM, Emre, NCT, Eng, CH, Engedal, N, Engelbrecht, A-M, Engelsen, AST, Enserink, JM, Escalante, R, Esclatine, A, Escobar-Henriques, M, Eskelinen, E-L, Espert, L, Eusebio, M-O, Fabrias, G, Fabrizi, C, Facchiano, A, Facchiano, F, Fadeel, B, Fader, C, Faesen, AC, Fairlie, WD, Falcó, A, Falkenburger, BH, Fan, D, Fan, J, Fan, Y, Fang, EF, Fang, Y, Fanto, M, Farfel-Becker, T, Faure, M, Fazeli, G, Fedele, AO, Feldman, AM, Feng, D, Feng, J, Feng, L, Feng, Y, Feng, W, Fenz Araujo, T, Ferguson, TA, Fernández, ÁF, Fernandez-Checa, JC, Fernández-Veledo, S, Fernie, AR, Ferrante, AW, Ferraresi, A, Ferrari, MF, Ferreira, JCB, Ferro-Novick, S, Figueras, A, Filadi, R, Filigheddu, N, Filippi-Chiela, E, Filomeni, G, Fimia, GM, Fineschi, V, Finetti, F, Finkbeiner, S, Fisher, EA, Fisher, PB, Flamigni, F, Fliesler, SJ, Flo, TH, Florance, I, Florey, O, Florio, T, Fodor, E, Follo, C, Fon, EA, Forlino, A, Fornai, F, Fortini, P, Fracassi, A, Fraldi, A, Franco, B, Franco, R, Franconi, F, Frankel, LB, Friedman, SL, Fröhlich, LF, Frühbeck, G, Fuentes, JM, Fujiki, Y, Fujita, N, Fujiwara, Y, Fukuda, M, Fulda, S, Furic, L, Furuya, N, Fusco, C, Gack, MU, Gaffke, L, Galadari, S, Galasso, A, Galindo, MF, Gallolu Kankanamalage, S, Galluzzi, L, Galy, V, Gammoh, N, Gan, B, Ganley, IG, Gao, F, Gao, H, Gao, M, Gao, P, Gao, S-J, Gao, W, Gao, X, Garcera, A, Garcia, MN, Garcia, VE, García-Del Portillo, F, Garcia-Escudero, V, Garcia-Garcia, A, Garcia-Macia, M, García-Moreno, D, Garcia-Ruiz, C, García-Sanz, P, Garg, AD, Gargini, R, Garofalo, T, Garry, RF, Gassen, NC, Gatica, D, Ge, L, Ge, W, Geiss-Friedlander, R, Gelfi, C, Genschik, P, Gentle, IE, Gerbino, V, Gerhardt, C, Germain, K, Germain, M, Gewirtz, DA, Ghasemipour Afshar, E, Ghavami, S, Ghigo, A, Ghosh, M, Giamas, G, Giampietri, C, Giatromanolaki, A, Gibson, GE, Gibson, SB, Ginet, V, Giniger, E, Giorgi, C, Girao, H, Girardin, SE, Giridharan, M, Giuliano, S, Giulivi, C, Giuriato, S, Giustiniani, J, Gluschko, A, Goder, V, Goginashvili, A, Golab, J, Goldstone, DC, Golebiewska, A, Gomes, LR, Gomez, R, Gómez-Sánchez, R, Gomez-Puerto, MC, Gomez-Sintes, R, Gong, Q, Goni, FM, González-Gallego, J, Gonzalez-Hernandez, T, Gonzalez-Polo, RA, Gonzalez-Reyes, JA, González-Rodríguez, P, Goping, IS, Gorbatyuk, MS, Gorbunov, NV, Görgülü, K, Gorojod, RM, Gorski, SM, Goruppi, S, Gotor, C, Gottlieb, RA, Gozes, I, Gozuacik, D, Graef, M, Gräler, MH, Granatiero, V, Grasso, D, Gray, JP, Green, DR, Greenhough, A, Gregory, SL, Griffin, EF, Grinstaff, MW, Gros, F, Grose, C, Gross, AS, Gruber, F, Grumati, P, Grune, T, Gu, X, Guan, J-L, Guardia, CM, Guda, K, Guerra, F, Guerri, C, Guha, P, Guillén, C, Gujar, S, Gukovskaya, A, Gukovsky, I, Gunst, J, Günther, A, Guntur, AR, Guo, C, Guo, H, Guo, L-W, Guo, M, Gupta, P, Gupta, SK, Gupta, S, Gupta, VB, Gupta, V, Gustafsson, AB, Gutterman, DD, H B, R, Haapasalo, A, Haber, JE, Hać, A, Hadano, S, Hafrén, AJ, Haidar, M, Hall, BS, Halldén, G, Hamacher-Brady, A, Hamann, A, Hamasaki, M, Han, W, Hansen, M, Hanson, PI, Hao, Z, Harada, M, Harhaji-Trajkovic, L, Hariharan, N, Haroon, N, Harris, J, Hasegawa, T, Hasima Nagoor, N, Haspel, JA, Haucke, V, Hawkins, WD, Hay, BA, Haynes, CM, Hayrabedyan, SB, Hays, TS, He, C, He, Q, He, R-R, He, Y-W, He, Y-Y, Heakal, Y, Heberle, AM, Hejtmancik, JF, Helgason, GV, Henkel, V, Herb, M, Hergovich, A, Herman-Antosiewicz, A, Hernández, A, Hernandez, C, Hernandez-Diaz, S, Hernandez-Gea, V, Herpin, A, Herreros, J, Hervás, JH, Hesselson, D, Hetz, C, Heussler, VT, Higuchi, Y, Hilfiker, S, Hill, JA, Hlavacek, WS, Ho, EA, Ho, IHT, Ho, PW-L, Ho, S-L, Ho, WY, Hobbs, GA, Hochstrasser, M, Hoet, PHM, Hofius, D, Hofman, P, Höhn, A, Holmberg, CI, Hombrebueno, JR, Yi-Ren Hong, C-WH, Hooper, LV, Hoppe, T, Horos, R, Hoshida, Y, Hsin, I-L, Hsu, H-Y, Hu, B, Hu, D, Hu, L-F, Hu, MC, Hu, R, Hu, W, Hu, Y-C, Hu, Z-W, Hua, F, Hua, J, Hua, Y, Huan, C, Huang, C, Huang, H, Huang, K, Huang, MLH, Huang, R, Huang, S, Huang, T, Huang, X, Huang, YJ, Huber, TB, Hubert, V, Hubner, CA, Hughes, SM, Hughes, WE, Humbert, M, Hummer, G, Hurley, JH, Hussain, S, Hussey, PJ, Hutabarat, M, Hwang, H-Y, Hwang, S, Ieni, A, Ikeda, F, Imagawa, Y, Imai, Y, Imbriano, C, Imoto, M, Inman, DM, Inoki, K, Iovanna, J, Iozzo, RV, Ippolito, G, Irazoqui, JE, Iribarren, P, Ishaq, M, Ishikawa, M, Ishimwe, N, Isidoro, C, Ismail, N, Issazadeh-Navikas, S, Itakura, E, Ito, D, Ivankovic, D, Ivanova, S, Iyer, AKV, Izquierdo, JM, Izumi, M, Jäättelä, M, Jabir, MS, Jackson, WT, Jacobo-Herrera, N, Jacomin, A-C, Jacquin, E, Jadiya, P, Jaeschke, H, Jagannath, C, Jakobi, AJ, Jakobsson, J, Janji, B, Jansen-Dürr, P, Jansson, PJ, Jantsch, J, Januszewski, S, Jassey, A, Jean, S, Jeltsch-David, H, Jendelova, P, Jenny, A, Jensen, TE, Jessen, N, Jewell, JL, Ji, J, Jia, L, Jia, R, Jiang, L, Jiang, Q, Jiang, R, Jiang, T, Jiang, X, Jiang, Y, Jimenez-Sanchez, M, Jin, E-J, Jin, F, Jin, H, Jin, L, Jin, M, Jin, S, Jo, E-K, Joffre, C, Johansen, T, Johnson, GVW, Johnston, SA, Jokitalo, E, Jolly, MK, Joosten, LAB, Jordan, J, Joseph, B, Ju, D, Ju, J-S, Ju, J, Juárez, E, Judith, D, Juhász, G, Jun, Y, Jung, CH, Jung, S-C, Jung, YK, Jungbluth, H, Jungverdorben, J, Just, S, Kaarniranta, K, Kaasik, A, Kabuta, T, Kaganovich, D, Kahana, A, Kain, R, Kajimura, S, Kalamvoki, M, Kalia, M, Kalinowski, DS, Kaludercic, N, Kalvari, I, Kaminska, J, Kaminskyy, VO, Kanamori, H, Kanasaki, K, Kang, C, Kang, R, Kang, SS, Kaniyappan, S, Kanki, T, Kanneganti, T-D, Kanthasamy, AG, Kanthasamy, A, Kantorow, M, Kapuy, O, Karamouzis, MV, Karim, MR, Karmakar, P, Katare, RG, Kato, M, Kaufmann, SHE, Kauppinen, A, Kaushal, GP, Kaushik, S, Kawasaki, K, Kazan, K, Ke, P-Y, Keating, DJ, Keber, U, Kehrl, JH, Keller, KE, Keller, CW, Kemper, JK, Kenific, CM, Kepp, O, Kermorgant, S, Kern, A, Ketteler, R, Keulers, TG, Khalfin, B, Khalil, H, Khambu, B, Khan, SY, Khandelwal, VKM, Khandia, R, Kho, W, Khobrekar, NV, Khuansuwan, S, Khundadze, M, Killackey, SA, Kim, D, Kim, DR, Kim, D-H, Kim, D-E, Kim, EY, Kim, E-K, Kim, H-R, Kim, H-S, Hyung-Ryong Kim, Kim, JH, Kim, JK, Kim, J-H, Kim, J, Kim, KI, Kim, PK, Kim, S-J, Kimball, SR, Kimchi, A, Kimmelman, AC, Kimura, T, King, MA, Kinghorn, KJ, Kinsey, CG, Kirkin, V, Kirshenbaum, LA, Kiselev, SL, Kishi, S, Kitamoto, K, Kitaoka, Y, Kitazato, K, Kitsis, RN, Kittler, JT, Kjaerulff, O, Klein, PS, Klopstock, T, Klucken, J, Knævelsrud, H, Knorr, RL, Ko, BCB, Ko, F, Ko, J-L, Kobayashi, H, Kobayashi, S, Koch, I, Koch, JC, Koenig, U, Kögel, D, Koh, YH, Koike, M, Kohlwein, SD, Kocaturk, NM, Komatsu, M, König, J, Kono, T, Kopp, BT, Korcsmaros, T, Korkmaz, G, Korolchuk, VI, Korsnes, MS, Koskela, A, Kota, J, Kotake, Y, Kotler, ML, Kou, Y, Koukourakis, MI, Koustas, E, Kovacs, AL, Kovács, T, Koya, D, Kozako, T, Kraft, C, Krainc, D, Krämer, H, Krasnodembskaya, AD, Kretz-Remy, C, Kroemer, G, Ktistakis, NT, Kuchitsu, K, Kuenen, S, Kuerschner, L, Kukar, T, Kumar, A, Kumar, D, Kumar, S, Kume, S, Kumsta, C, Kundu, CN, Kundu, M, Kunnumakkara, AB, Kurgan, L, Kutateladze, TG, Kutlu, O, Kwak, S, Kwon, HJ, Kwon, TK, Kwon, YT, Kyrmizi, I, La Spada, A, Labonté, P, Ladoire, S, Laface, I, Lafont, F, Lagace, DC, Lahiri, V, Lai, Z, Laird, AS, Lakkaraju, A, Lamark, T, Lan, S-H, Landajuela, A, Lane, DJR, Lane, JD, Lang, CH, Lange, C, Langel, Ü, Langer, R, Lapaquette, P, Laporte, J, LaRusso, NF, Lastres-Becker, I, Lau, WCY, Laurie, GW, Lavandero, S, Law, BYK, Law, HK-W, Layfield, R, Le, W, Le Stunff, H, Leary, AY, Lebrun, J-J, Leck, LYW, Leduc-Gaudet, J-P, Lee, C, Lee, C-P, Lee, D-H, Lee, EB, Lee, EF, Lee, GM, Lee, H-J, Lee, HK, Lee, JM, Lee, JS, Lee, J-A, Lee, J-Y, Lee, JH, Lee, M, Lee, MG, Lee, MJ, Lee, M-S, Lee, SY, Lee, S-J, Lee, SB, Lee, WH, Lee, Y-R, Lee, Y-H, Lee, Y, Lefebvre, C, Legouis, R, Lei, YL, Lei, Y, Leikin, S, Leitinger, G, Lemus, L, Leng, S, Lenoir, O, Lenz, G, Lenz, HJ, Lenzi, P, León, Y, Leopoldino, AM, Leschczyk, C, Leskelä, S, Letellier, E, Leung, C-T, Leung, PS, Leventhal, JS, Levine, B, Lewis, PA, Ley, K, Li, B, Li, D-Q, Li, J, Li, K, Li, L, Li, M, Li, P-L, Li, M-Q, Li, Q, Li, S, Li, T, Li, W, Li, X, Li, Y-P, Li, Y, Li, Z, Lian, J, Liang, C, Liang, Q, Liang, W, Liang, Y, Liao, G, Liao, L, Liao, M, Liao, Y-F, Librizzi, M, Lie, PPY, Lilly, MA, Lim, HJ, Lima, TRR, Limana, F, Lin, C, Lin, C-W, Lin, D-S, Lin, F-C, Lin, JD, Lin, KM, Lin, K-H, Lin, L-T, Lin, P-H, Lin, Q, Lin, S, Lin, S-J, Lin, W, Lin, X, Lin, Y-X, Lin, Y-S, Linden, R, Lindner, P, Ling, S-C, Lingor, P, Linnemann, AK, Liou, Y-C, Lipinski, MM, Lipovšek, S, Lira, VA, Lisiak, N, Liton, PB, Liu, C, Liu, C-H, Liu, C-F, Liu, CH, Liu, F, Liu, H, Liu, H-S, Liu, H-F, Liu, J, Liu, L, Liu, M, Liu, Q, Liu, W, Liu, X-H, Liu, X, Liu, Y, Livingston, JA, Lizard, G, Lizcano, JM, Ljubojevic-Holzer, S, LLeonart, ME, Llobet-Navàs, D, Llorente, A, Lo, CH, Lobato-Márquez, D, Long, Q, Long, YC, Loos, B, Loos, JA, López, MG, López-Doménech, G, López-Guerrero, JA, López-Jiménez, AT, López-Pérez, Ó, López-Valero, I, Lorenowicz, MJ, Lorente, M, Lorincz, P, Lossi, L, Lotersztajn, S, Lovat, PE, Lovell, JF, Lovy, A, Lőw, P, Lu, G, Lu, H, Lu, J-H, Lu, J-J, Lu, M, Lu, S, Luciani, A, Lucocq, JM, Ludovico, P, Luftig, MA, Luhr, M, Luis-Ravelo, D, Lum, JJ, Luna-Dulcey, L, Lund, AH, Lund, VK, Lünemann, JD, Lüningschrör, P, Luo, H, Luo, R, Luo, S, Luo, Z, Luparello, C, Lüscher, B, Luu, L, Lyakhovich, A, Lyamzaev, KG, Lystad, AH, Lytvynchuk, L, Ma, AC, Ma, C, Ma, M, Ma, N-F, Ma, Q-H, Ma, X, Ma, Y, Ma, Z, MacDougald, OA, Macian, F, MacIntosh, GC, MacKeigan, JP, Macleod, KF, Maday, S, Madeo, F, Madesh, M, Madl, T, Madrigal-Matute, J, Maeda, A, Maejima, Y, Magarinos, M, Mahavadi, P, Maiani, E, Maiese, K, Maiti, P, Maiuri, MC, Majello, B, Major, MB, Makareeva, E, Malik, F, Mallilankaraman, K, Malorni, W, Maloyan, A, Mammadova, N, Man, GCW, Manai, F, Mancias, JD, Mandelkow, E-M, Mandell, MA, Manfredi, AA, Manjili, MH, Manjithaya, R, Manque, P, Manshian, BB, Manzano, R, Manzoni, C, Mao, K, Marchese, C, Marchetti, S, Marconi, AM, Marcucci, F, Mardente, S, Mareninova, OA, Margeta, M, Mari, M, Marinelli, S, Marinelli, O, Mariño, G, Mariotto, S, Marshall, RS, Marten, MR, Martens, S, Martin, APJ, Martin, KR, Martin, S, Martín-Segura, A, Martín-Acebes, MA, Martin-Burriel, I, Martin-Rincon, M, Martin-Sanz, P, Martina, JA, Martinet, W, Martinez, A, Martinez, J, Martinez Velazquez, M, Martinez-Lopez, N, Martinez-Vicente, M, Martins, DO, Martins, JO, Martins, WK, Martins-Marques, T, Marzetti, E, Masaldan, S, Masclaux-Daubresse, C, Mashek, DG, Massa, V, Massieu, L, Masson, GR, Masuelli, L, Masyuk, AI, Masyuk, TV, Matarrese, P, Matheu, A, Matoba, S, Matsuzaki, S, Mattar, P, Matte, A, Mattoscio, D, Mauriz, JL, Mauthe, M, Mauvezin, C, Maverakis, E, Maycotte, P, Mayer, J, Mazzoccoli, G, Mazzoni, C, Mazzulli, JR, McCarty, N, McDonald, C, McGill, MR, McKenna, SL, McLaughlin, B, McLoughlin, F, McNiven, MA, McWilliams, TG, Mechta-Grigoriou, F, Medeiros, TC, Medina, DL, Megeney, LA, Megyeri, K, Mehrpour, M, Mehta, JL, Meijer, AJ, Meijer, AH, Mejlvang, J, Meléndez, A, Melk, A, Memisoglu, G, Mendes, AF, Meng, D, Meng, F, Meng, T, Menna-Barreto, R, Menon, MB, Mercer, C, Mercier, AE, Mergny, J-L, Merighi, A, Merkley, SD, Merla, G, Meske, V, Mestre, AC, Metur, SP, Meyer, C, Meyer, H, Mi, W, Mialet-Perez, J, Miao, J, Micale, L, Miki, Y, Milan, E, Milczarek, M, Miller, DL, Miller, SI, Miller, S, Millward, SW, Milosevic, I, Minina, EA, Mirzaei, H, Mirzaei, HR, Mirzaei, M, Mishra, A, Mishra, N, Mishra, PK, Misirkic Marjanovic, M, Misasi, R, Misra, A, Misso, G, Mitchell, C, Mitou, G, Miura, T, Miyamoto, S, Miyazaki, M, Miyazaki, T, Miyazawa, K, Mizushima, N, Mogensen, TH, Mograbi, B, Mohammadinejad, R, Mohamud, Y, Mohanty, A, Mohapatra, S, Möhlmann, T, Mohmmed, A, Moles, A, Moley, KH, Molinari, M, Mollace, V, Møller, AB, Mollereau, B, Mollinedo, F, Montagna, C, Monteiro, MJ, Montella, A, Montes, LR, Montico, B, Mony, VK, Monzio Compagnoni, G, Moore, MN, Moosavi, MA, Mora, AL, Mora, M, Morales-Alamo, D, Moratalla, R, Moreira, PI, Morelli, E, Moreno, S, Moreno-Blas, D, Moresi, V, Morga, B, Morgan, AH, Morin, F, Morishita, H, Moritz, OL, Moriyama, M, Moriyasu, Y, Morleo, M, Morselli, E, Moruno-Manchon, JF, Moscat, J, Mostowy, S, Motori, E, Moura, AF, Moustaid-Moussa, N, Mrakovcic, M, Muciño-Hernández, G, Mukherjee, A, Mukhopadhyay, S, Mulcahy Levy, JM, Mulero, V, Muller, S, Münch, C, Munjal, A, Munoz-Canoves, P, Muñoz-Galdeano, T, Münz, C, Murakawa, T, Muratori, C, Murphy, BM, Murphy, JP, Murthy, A, Myöhänen, TT, Mysorekar, IU, Mytych, J, Nabavi, SM, Nabissi, M, Nagy, P, Nah, J, Nahimana, A, Nakagawa, I, Nakamura, K, Nakatogawa, H, Nandi, SS, Nanjundan, M, Nanni, M, Napolitano, G, Nardacci, R, Narita, M, Nassif, M, Nathan, I, Natsumeda, M, Naude, RJ, Naumann, C, Naveiras, O, Navid, F, Nawrocki, ST, Nazarko, TY, Nazio, F, Negoita, F, Neill, T, Neisch, AL, Neri, LM, Netea, MG, Neubert, P, Neufeld, TP, Neumann, D, Neutzner, A, Newton, PT, Ney, PA, Nezis, IP, Ng, CCW, Ng, TB, Nguyen, HTT, Nguyen, LT, Ni, H-M, Ní Cheallaigh, C, Ni, Z, Nicolao, MC, Nicoli, F, Nieto-Diaz, M, Nilsson, P, Ning, S, Niranjan, R, Nishimune, H, Niso-Santano, M, Nixon, RA, Nobili, A, Nobrega, C, Noda, T, Nogueira-Recalde, U, Nolan, TM, Nombela, I, Novak, I, Novoa, B, Nozawa, T, Nukina, N, Nussbaum-Krammer, C, Nylandsted, J, O'Donovan, TR, O'Leary, SM, O'Rourke, EJ, O'Sullivan, MP, O'Sullivan, TE, Oddo, S, Oehme, I, Ogawa, M, Ogier-Denis, E, Ogmundsdottir, MH, Ogretmen, B, Oh, GT, Oh, S-H, Oh, YJ, Ohama, T, Ohashi, Y, Ohmuraya, M, Oikonomou, V, Ojha, R, Okamoto, K, Okazawa, H, Oku, M, Oliván, S, Oliveira, JMA, Ollmann, M, Olzmann, JA, Omari, S, Omary, MB, Önal, G, Ondrej, M, Ong, S-B, Ong, S-G, Onnis, A, Orellana, JA, Orellana-Muñoz, S, Ortega-Villaizan, MDM, Ortiz-Gonzalez, XR, Ortona, E, Osiewacz, HD, Osman, A-HK, Osta, R, Otegui, MS, Otsu, K, Ott, C, Ottobrini, L, Ou, J-HJ, Outeiro, TF, Oynebraten, I, Ozturk, M, Pagès, G, Pahari, S, Pajares, M, Pajvani, UB, Pal, R, Paladino, S, Pallet, N, Palmieri, M, Palmisano, G, Palumbo, C, Pampaloni, F, Pan, L, Pan, Q, Pan, W, Pan, X, Panasyuk, G, Pandey, R, Pandey, UB, Pandya, V, Paneni, F, Pang, SY, Panzarini, E, Papademetrio, DL, Papaleo, E, Papinski, D, Papp, D, Park, EC, Park, HT, Park, J-M, Park, J-I, Park, JT, Park, J, Park, SC, Park, S-Y, Parola, AH, Parys, JB, Pasquier, A, Pasquier, B, Passos, JF, Pastore, N, Patel, HH, Patschan, D, Pattingre, S, Pedraza-Alva, G, Pedraza-Chaverri, J, Pedrozo, Z, Pei, G, Pei, J, Peled-Zehavi, H, Pellegrini, JM, Pelletier, J, Peñalva, MA, Peng, D, Peng, Y, Penna, F, Pennuto, M, Pentimalli, F, Pereira, CM, Pereira, GJS, Pereira, LC, Pereira de Almeida, L, Perera, ND, Pérez-Lara, Á, Perez-Oliva, AB, Pérez-Pérez, ME, Periyasamy, P, Perl, A, Perrotta, C, Perrotta, I, Pestell, RG, Petersen, M, Petrache, I, Petrovski, G, Pfirrmann, T, Pfister, AS, Philips, JA, Pi, H, Picca, A, Pickrell, AM, Picot, S, Pierantoni, GM, Pierdominici, M, Pierre, P, Pierrefite-Carle, V, Pierzynowska, K, Pietrocola, F, Pietruczuk, M, Pignata, C, Pimentel-Muiños, FX, Pinar, M, Pinheiro, RO, Pinkas-Kramarski, R, Pinton, P, Pircs, K, Piya, S, Pizzo, P, Plantinga, TS, Platta, HW, Plaza-Zabala, A, Plomann, M, Plotnikov, EY, Plun-Favreau, H, Pluta, R, Pocock, R, Pöggeler, S, Pohl, C, Poirot, M, Poletti, A, Ponpuak, M, Popelka, H, Popova, B, Porta, H, Porte Alcon, S, Portilla-Fernandez, E, Post, M, Potts, MB, Poulton, J, Powers, T, Prahlad, V, Prajsnar, TK, Praticò, D, Prencipe, R, Priault, M, Proikas-Cezanne, T, Promponas, VJ, Proud, CG, Puertollano, R, Puglielli, L, Pulinilkunnil, T, Puri, D, Puri, R, Puyal, J, Qi, X, Qi, Y, Qian, W, Qiang, L, Qiu, Y, Quadrilatero, J, Quarleri, J, Raben, N, Rabinowich, H, Ragona, D, Ragusa, MJ, Rahimi, N, Rahmati, M, Raia, V, Raimundo, N, Rajasekaran, N-S, Ramachandra Rao, S, Rami, A, Ramírez-Pardo, I, Ramsden, DB, Randow, F, Rangarajan, PN, Ranieri, D, Rao, H, Rao, L, Rao, R, Rathore, S, Ratnayaka, JA, Ratovitski, EA, Ravanan, P, Ravegnini, G, Ray, SK, Razani, B, Rebecca, V, Reggiori, F, Régnier-Vigouroux, A, Reichert, AS, Reigada, D, Reiling, JH, Rein, T, Reipert, S, Rekha, RS, Ren, H, Ren, J, Ren, W, Renault, T, Renga, G, Reue, K, Rewitz, K, Ribeiro de Andrade Ramos, B, Riazuddin, SA, Ribeiro-Rodrigues, TM, Ricci, J-E, Ricci, R, Riccio, V, Richardson, DR, Rikihisa, Y, Risbud, MV, Risueño, RM, Ritis, K, Rizza, S, Rizzuto, R, Roberts, HC, Roberts, LD, Robinson, KJ, Roccheri, MC, Rocchi, S, Rodney, GG, Rodrigues, T, Rodrigues Silva, VR, Rodriguez, A, Rodriguez-Barrueco, R, Rodriguez-Henche, N, Rodriguez-Rocha, H, Roelofs, J, Rogers, RS, Rogov, VV, Rojo, AI, Rolka, K, Romanello, V, Romani, L, Romano, A, Romano, PS, Romeo-Guitart, D, Romero, LC, Romero, M, Roney, JC, Rongo, C, Roperto, S, Rosenfeldt, MT, Rosenstiel, P, Rosenwald, AG, Roth, KA, Roth, L, Roth, S, Rouschop, KMA, Roussel, BD, Roux, S, Rovere-Querini, P, Roy, A, Rozieres, A, Ruano, D, Rubinsztein, DC, Rubtsova, MP, Ruckdeschel, K, Ruckenstuhl, C, Rudolf, E, Rudolf, R, Ruggieri, A, Ruparelia, AA, Rusmini, P, Russell, RR, Russo, GL, Russo, M, Russo, R, Ryabaya, OO, Ryan, KM, Ryu, K-Y, Sabater-Arcis, M, Sachdev, U, Sacher, M, Sachse, C, Sadhu, A, Sadoshima, J, Safren, N, Saftig, P, Sagona, AP, Sahay, G, Sahebkar, A, Sahin, M, Sahin, O, Sahni, S, Saito, N, Saito, S, Saito, T, Sakai, R, Sakai, Y, Sakamaki, J-I, Saksela, K, Salazar, G, Salazar-Degracia, A, Salekdeh, GH, Saluja, AK, Sampaio-Marques, B, Sanchez, MC, Sanchez-Alcazar, JA, Sanchez-Vera, V, Sancho-Shimizu, V, Sanderson, JT, Sandri, M, Santaguida, S, Santambrogio, L, Santana, MM, Santoni, G, Sanz, A, Sanz, P, Saran, S, Sardiello, M, Sargeant, TJ, Sarin, A, Sarkar, C, Sarkar, S, Sarrias, M-R, Sarmah, DT, Sarparanta, J, Sathyanarayan, A, Sathyanarayanan, R, Scaglione, KM, Scatozza, F, Schaefer, L, Schafer, ZT, Schaible, UE, Schapira, AHV, Scharl, M, Schatzl, HM, Schein, CH, Scheper, W, Scheuring, D, Schiaffino, MV, Schiappacassi, M, Schindl, R, Schlattner, U, Schmidt, O, Schmitt, R, Schmidt, SD, Schmitz, I, Schmukler, E, Schneider, A, Schneider, BE, Schober, R, Schoijet, AC, Schott, MB, Schramm, M, Schröder, B, Schuh, K, Schüller, C, Schulze, RJ, Schürmanns, L, Schwamborn, JC, Schwarten, M, Scialo, F, Sciarretta, S, Scott, MJ, Scotto, KW, Scovassi, AI, Scrima, A, Scrivo, A, Sebastian, D, Sebti, S, Sedej, S, Segatori, L, Segev, N, Seglen, PO, Seiliez, I, Seki, E, Selleck, SB, Sellke, FW, Selsby, JT, Sendtner, M, Senturk, S, Seranova, E, Sergi, C, Serra-Moreno, R, Sesaki, H, Settembre, C, Setty, SRG, Sgarbi, G, Sha, O, Shacka, JJ, Shah, JA, Shang, D, Shao, C, Shao, F, Sharbati, S, Sharkey, LM, Sharma, D, Sharma, G, Sharma, K, Sharma, P, Sharma, S, Shen, H-M, Shen, H, Shen, J, Shen, M, Shen, W, Shen, Z, Sheng, R, Sheng, Z, Sheng, Z-H, Shi, J, Shi, X, Shi, Y-H, Shiba-Fukushima, K, Shieh, J-J, Shimada, Y, Shimizu, S, Shimozawa, M, Shintani, T, Shoemaker, CJ, Shojaei, S, Shoji, I, Shravage, BV, Shridhar, V, Shu, C-W, Shu, H-B, Shui, K, Shukla, AK, Shutt, TE, Sica, V, Siddiqui, A, Sierra, A, Sierra-Torre, V, Signorelli, S, Sil, P, Silva, BJDA, Silva, JD, Silva-Pavez, E, Silvente-Poirot, S, Simmonds, RE, Simon, AK, Simon, H-U, Simons, M, Singh, A, Singh, LP, Singh, R, Singh, SV, Singh, SK, Singh, SB, Singh, S, Singh, SP, Sinha, D, Sinha, RA, Sinha, S, Sirko, A, Sirohi, K, Sivridis, EL, Skendros, P, Skirycz, A, Slaninová, I, Smaili, SS, Smertenko, A, Smith, MD, Soenen, SJ, Sohn, EJ, Sok, SPM, Solaini, G, Soldati, T, Soleimanpour, SA, Soler, RM, Solovchenko, A, Somarelli, JA, Sonawane, A, Song, F, Song, HK, Song, J-X, Song, K, Song, Z, Soria, LR, Sorice, M, Soukas, AA, Soukup, S-F, Sousa, D, Sousa, N, Spagnuolo, PA, Spector, SA, Srinivas Bharath, MM, St Clair, D, Stagni, V, Staiano, L, Stalnecker, CA, Stankov, MV, Stathopulos, PB, Stefan, K, Stefan, SM, Stefanis, L, Steffan, JS, Steinkasserer, A, Stenmark, H, Sterneckert, J, Stevens, C, Stoka, V, Storch, S, Stork, B, Strappazzon, F, Strohecker, AM, Stupack, DG, Su, H, Su, L-Y, Su, L, Suarez-Fontes, AM, Subauste, CS, Subbian, S, Subirada, PV, Sudhandiran, G, Sue, CM, Sui, X, Summers, C, Sun, G, Sun, J, Sun, K, Sun, M-X, Sun, Q, Sun, Y, Sun, Z, Sunahara, KKS, Sundberg, E, Susztak, K, Sutovsky, P, Suzuki, H, Sweeney, G, Symons, JD, Sze, SCW, Szewczyk, NJ, Tabęcka-Łonczynska, A, Tabolacci, C, Tacke, F, Taegtmeyer, H, Tafani, M, Tagaya, M, Tai, H, Tait, SWG, Takahashi, Y, Takats, S, Talwar, P, Tam, C, Tam, SY, Tampellini, D, Tamura, A, Tan, CT, Tan, E-K, Tan, Y-Q, Tanaka, M, Tang, D, Tang, J, Tang, T-S, Tanida, I, Tao, Z, Taouis, M, Tatenhorst, L, Tavernarakis, N, Taylor, A, Taylor, GA, Taylor, JM, Tchetina, E, Tee, AR, Tegeder, I, Teis, D, Teixeira, N, Teixeira-Clerc, F, Tekirdag, KA, Tencomnao, T, Tenreiro, S, Tepikin, AV, Testillano, PS, Tettamanti, G, Tharaux, P-L, Thedieck, K, Thekkinghat, AA, Thellung, S, Thinwa, JW, Thirumalaikumar, VP, Thomas, SM, Thomes, PG, Thorburn, A, Thukral, L, Thum, T, Thumm, M, Tian, L, Tichy, A, Till, A, Timmerman, V, Titorenko, VI, Todi, SV, Todorova, K, Toivonen, JM, Tomaipitinca, L, Tomar, D, Tomas-Zapico, C, Tomić, S, Tong, BC-K, Tong, C, Tong, X, Tooze, SA, Torgersen, ML, Torii, S, Torres-López, L, Torriglia, A, Towers, CG, Towns, R, Toyokuni, S, Trajkovic, V, Tramontano, D, Tran, Q-G, Travassos, LH, Trelford, CB, Tremel, S, Trougakos, IP, Tsao, BP, Tschan, MP, Tse, H-F, Tse, TF, Tsugawa, H, Tsvetkov, AS, Tumbarello, DA, Tumtas, Y, Tuñón, MJ, Turcotte, S, Turk, B, Turk, V, Turner, BJ, Tuxworth, RI, Tyler, JK, Tyutereva, EV, Uchiyama, Y, Ugun-Klusek, A, Uhlig, HH, Ułamek-Kozioł, M, Ulasov, IV, Umekawa, M, Ungermann, C, Unno, R, Urbe, S, Uribe-Carretero, E, Üstün, S, Uversky, VN, Vaccari, T, Vaccaro, MI, Vahsen, BF, Vakifahmetoglu-Norberg, H, Valdor, R, Valente, MJ, Valko, A, Vallee, RB, Valverde, AM, Van den Berghe, G, van der Veen, S, Van Kaer, L, van Loosdregt, J, van Wijk, SJL, Vandenberghe, W, Vanhorebeek, I, Vannier-Santos, MA, Vannini, N, Vanrell, MC, Vantaggiato, C, Varano, G, Varela-Nieto, I, Varga, M, Vasconcelos, MH, Vats, S, Vavvas, DG, Vega-Naredo, I, Vega-Rubin-de-Celis, S, Velasco, G, Velázquez, AP, Vellai, T, Vellenga, E, Velotti, F, Verdier, M, Verginis, P, Vergne, I, Verkade, P, Verma, M, Verstreken, P, Vervliet, T, Vervoorts, J, Vessoni, AT, Victor, VM, Vidal, M, Vidoni, C, Vieira, OV, Vierstra, RD, Viganó, S, Vihinen, H, Vijayan, V, Vila, M, Vilar, M, Villalba, JM, Villalobo, A, Villarejo-Zori, B, Villarroya, F, Villarroya, J, Vincent, O, Vindis, C, Viret, C, Viscomi, MT, Visnjic, D, Vitale, I, Vocadlo, DJ, Voitsekhovskaja, OV, Volonté, C, Volta, M, Vomero, M, Von Haefen, C, Vooijs, MA, Voos, W, Vucicevic, L, Wade-Martins, R, Waguri, S, Waite, KA, Wakatsuki, S, Walker, DW, Walker, MJ, Walker, SA, Walter, J, Wandosell, FG, Wang, B, Wang, C-Y, Wang, C, Wang, D, Wang, F, Wang, G, Wang, H, Wang, H-G, Wang, J, Wang, K, Wang, L, Wang, MH, Wang, M, Wang, N, Wang, P, Wang, QJ, Wang, Q, Wang, QK, Wang, QA, Wang, W-T, Wang, W, Wang, X, Wang, Y, Wang, Y-Y, Wang, Z, Warnes, G, Warnsmann, V, Watada, H, Watanabe, E, Watchon, M, Wawrzyńska, A, Weaver, TE, Wegrzyn, G, Wehman, AM, Wei, H, Wei, L, Wei, T, Wei, Y, Weiergräber, OH, Weihl, CC, Weindl, G, Weiskirchen, R, Wells, A, Wen, RH, Wen, X, Werner, A, Weykopf, B, Wheatley, SP, Whitton, JL, Whitworth, AJ, Wiktorska, K, Wildenberg, ME, Wileman, T, Wilkinson, S, Willbold, D, Williams, B, Williams, RSB, Williams, RL, Williamson, PR, Wilson, RA, Winner, B, Winsor, NJ, Witkin, SS, Wodrich, H, Woehlbier, U, Wollert, T, Wong, E, Wong, JH, Wong, RW, Wong, VKW, Wong, WW-L, Wu, A-G, Wu, C, Wu, J, Wu, KK, Wu, M, Wu, S-Y, Wu, S, Wu, WKK, Wu, X, Wu, Y-W, Wu, Y, Xavier, RJ, Xia, H, Xia, L, Xia, Z, Xiang, G, Xiang, J, Xiang, M, Xiang, W, Xiao, B, Xiao, G, Xiao, H, Xiao, H-T, Xiao, J, Xiao, L, Xiao, S, Xiao, Y, Xie, B, Xie, C-M, Xie, M, Xie, Y, Xie, Z, Xilouri, M, Xu, C, Xu, E, Xu, H, Xu, J, Xu, L, Xu, WW, Xu, X, Xue, Y, Yakhine-Diop, SMS, Yamaguchi, M, Yamaguchi, O, Yamamoto, A, Yamashina, S, Yan, S, Yan, S-J, Yan, Z, Yanagi, Y, Yang, C, Yang, D-S, Yang, H, Yang, H-T, Yang, J-M, Yang, J, Yang, L, Yang, M, Yang, P-M, Yang, Q, Yang, S, Yang, S-F, Yang, W, Yang, WY, Yang, X, Yang, Y, Yao, H, Yao, S, Yao, X, Yao, Y-G, Yao, Y-M, Yasui, T, Yazdankhah, M, Yen, PM, Yi, C, Yin, X-M, Yin, Y, Yin, Z, Ying, M, Ying, Z, Yip, CK, Yiu, SPT, Yoo, YH, Yoshida, K, Yoshii, SR, Yoshimori, T, Yousefi, B, Yu, B, Yu, H, Yu, J, Yu, L, Yu, M-L, Yu, S-W, Yu, VC, Yu, WH, Yu, Z, Yuan, J, Yuan, L-Q, Yuan, S, Yuan, S-SF, Yuan, Y, Yuan, Z, Yue, J, Yue, Z, Yun, J, Yung, RL, Zacks, DN, Zaffagnini, G, Zambelli, VO, Zanella, I, Zang, QS, Zanivan, S, Zappavigna, S, Zaragoza, P, Zarbalis, KS, Zarebkohan, A, Zarrouk, A, Zeitlin, SO, Zeng, J, Zeng, J-D, Žerovnik, E, Zhan, L, Zhang, B, Zhang, DD, Zhang, H, Zhang, H-L, Zhang, J, Zhang, J-P, Zhang, KYB, Zhang, LW, Zhang, L, Zhang, M, Zhang, P, Zhang, S, Zhang, W, Zhang, X, Zhang, X-W, Zhang, XD, Zhang, Y, Zhang, Y-D, Zhang, Y-Y, Zhang, Z, Zhao, H, Zhao, L, Zhao, S, Zhao, T, Zhao, X-F, Zhao, Y, Zheng, G, Zheng, K, Zheng, L, Zheng, S, Zheng, X-L, Zheng, Y, Zheng, Z-G, Zhivotovsky, B, Zhong, Q, Zhou, A, Zhou, B, Zhou, C, Zhou, G, Zhou, H, Zhou, J, Zhou, K, Zhou, R, Zhou, X-J, Zhou, Y, Zhou, Z-Y, Zhou, Z, Zhu, B, Zhu, C, Zhu, G-Q, Zhu, H, Zhu, W-G, Zhu, Y, Zhuang, H, Zhuang, X, Zientara-Rytter, K, Zimmermann, CM, Ziviani, E, Zoladek, T, Zong, W-X, Zorov, DB, Zorzano, A, Zou, W, Zou, Z, Zuryn, S, Zwerschke, W, Brand-Saberi, B, Dong, XC, Kenchappa, CS, Lin, Y, Oshima, S, Rong, Y, Sluimer, JC, Stallings, CL, and Tong, C-K
- Abstract
In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
- Published
- 2021
3. Dissecting Functional, Structural, and Molecular Requirements for Serotonin Release from Mouse Enterochromaffin Cells
- Author
-
Shaaban, A, Maaß, F, Schwarze, V, Lund, ML, Beuermann, S, Chan, M, Harenberg, C, Bewick, GA, Keating, DJ, Benseler, F, Cooper, BH, Imig, C, Shaaban, A, Maaß, F, Schwarze, V, Lund, ML, Beuermann, S, Chan, M, Harenberg, C, Bewick, GA, Keating, DJ, Benseler, F, Cooper, BH, and Imig, C
- Published
- 2021
4. The gut microbiome regulates host glucose homeostasis via peripheral serotonin.
- Author
-
Martin, AM, Yabut, JM, Choo, JM, Page, AJ, Sun, EW, Jessup, CF, Wesselingh, SL, Khan, WI, Rogers, GB, Steinberg, GR, Keating, DJ, Martin, AM, Yabut, JM, Choo, JM, Page, AJ, Sun, EW, Jessup, CF, Wesselingh, SL, Khan, WI, Rogers, GB, Steinberg, GR, and Keating, DJ
- Abstract
The gut microbiome is an established regulator of aspects of host metabolism, such as glucose handling. Despite the known impacts of the gut microbiota on host glucose homeostasis, the underlying mechanisms are unknown. The gut microbiome is also a potent mediator of gut-derived serotonin synthesis, and this peripheral source of serotonin is itself a regulator of glucose homeostasis. Here, we determined whether the gut microbiome influences glucose homeostasis through effects on gut-derived serotonin. Using both pharmacological inhibition and genetic deletion of gut-derived serotonin synthesis, we find that the improvements in host glucose handling caused by antibiotic-induced changes in microbiota composition are dependent on the synthesis of peripheral serotonin.
- Published
- 2019
5. The nutrient-sensing repertoires of mouse enterochromaffin cells differ between duodenum and colon
- Author
-
Amanda L. Lumsden, Alyce M. Martin, Claire F. Jessup, Damien J. Keating, Richard L. Young, Nick J. Spencer, Martin, AM, Lumsden, AL, Young, RL, Jessup, CF, Spencer, NJ, and Keating, DJ
- Subjects
0301 basic medicine ,Cell physiology ,Male ,Physiology ,Colon ,Duodenum ,receptors ,Motility ,Gene Expression ,Nutrient sensing ,Biology ,enterochromaffin ,Receptors, G-Protein-Coupled ,03 medical and health sciences ,nutrients ,Free fatty acid receptor 1 ,Enterochromaffin Cells ,Animals ,glucose ,Receptor ,chemistry.chemical_classification ,Endocrine and Autonomic Systems ,Gastroenterology ,Fatty acid ,serotonin ,030104 developmental biology ,Biochemistry ,chemistry ,Enterochromaffin cell ,Mice, Inbred CBA ,Serotonin - Abstract
Background: Enterochromaffin (EC) cells within the gastrointestinal (GI) tract provide almost all body serotonin (5-hydroxytryptamine [5-HT]). Peripheral 5-HT, released from EC cells lining the gut wall, serves diverse physiological roles. These include modulating GI motility, bone formation, hepatic gluconeogenesis, thermogenesis, insulin resistance, and regulation of fat mass. Enterochromaffin cells are nutrient sensors, but which nutrients they are responsive to and how this changes in different parts of the GI tract are poorly understood. Methods: To accurately undertake such an examination, we undertook the first isolation and purification of primary mouse EC cells from both the duodenum and colon in the same animal. This allowed us to compare, in an internally controlled manner, regional differences in the expression of nutrient sensors in EC cells using real-time PCR. Key Results: Both colonic and duodenal EC cells expressed G protein-coupled receptors and facilitative transporters for sugars, free fatty acids, amino acids, and lipid amides. We find differential expression of nutrient receptor and transporters in EC cells obtained from duodenal and colonic EC cells. Duodenal EC cells have higher expression of tryptophan hydroxylase-1, sugar transporters GLUT2, GLUT5, and free fatty acid receptors 1 and 3 (FFAR1 and FFAR3). Colonic EC cells express higher levels of GLUT1, FFAR2, and FFAR4. Conclusions & Inferences: We highlight the diversity of EC cell physiology and identify differences in the regional sensing repertoire of EC cells to an assortment of nutrients. These data indicate that not all EC cells are similar and that differences in their physiological responses are likely dependent on their location within the GI tract. Refereed/Peer-reviewed
- Published
- 2016
6. Exocytosis is impaired in mucopolysaccharidosis IIIA mouse chromaffin cells
- Author
-
Doug A. Brooks, Damien J. Keating, E.H. Teo, John J. Hopwood, Kim M. Hemsley, Marnie Winter, Emma J. Parkinson-Lawrence, Kimberly D. Mackenzie, Keating, DJ, Winter, MA, Hemsley, KM, Mackenzie, KD, Teo, EH, Hopwood, JJ, Brooks, DA, and Parkinson-Lawrence, EJ
- Subjects
medicine.medical_specialty ,Chromaffin Cells ,Mucopolysaccharidosis ,Mice, Transgenic ,Stimulation ,Neurotransmission ,Biology ,Exocytosis ,Statistics, Nonparametric ,carbon fibre amperometry ,Mice ,Mucopolysaccharidosis III ,chemistry.chemical_compound ,Catecholamines ,Microscopy, Electron, Transmission ,Carbon Fiber ,Internal medicine ,Adrenal Glands ,medicine ,Animals ,Secretion ,neurotransmission ,chromaffin cells ,Neurotransmitter ,Cells, Cultured ,mouse ,Analysis of Variance ,General Neuroscience ,medicine.disease ,Carbon ,mucopolysaccharidosis IIIA ,Mice, Inbred C57BL ,Disease Models, Animal ,Endocrinology ,chemistry ,Catecholamine ,Calcium ,Heparitin Sulfate ,Lysosomes ,exocytosis ,medicine.drug - Abstract
Mucopolysaccharidosis IIIA (MPS IIIA) is a lysosomal storage disorder caused by a deficiency in the activity of the lysosomal hydrolase, sulphamidase, an enzyme involved in the degradation of heparan sulphate. MPS IIIA patients exhibit progressive mental retardation and behavioural disturbance. While neuropathology is the major clinical problem in MPS IIIA patients, there is little understanding of how lysosomal storage generates this phenotype. As reduced neuronal communication can underlie cognitive deficiencies, we investigated whether the secretion of neurotransmitters is altered in MPS IIIA mice; utilising adrenal chromaffin cells, a classical model for studying secretion via exocytosis. MPS IIIA chromaffin cells displayed heparan sulphate storage and electron microscopy revealed large electron-lucent storage compartments. There were also increased numbers of large/elongated chromaffin granules, with a morphology that was similar to immature secretory granules. Carbon fibre amperometry illustrated a significant decrease in the number of exocytotic events for MPS IIIA, when compared to control chromaffin cells. However, there were no changes in the kinetics of release, the amount of catecholamine released per exocytoticevent, or the amount of Ca2+ entry upon stimulation.The increased number of large/elongated granules andreduced number of exocytotic events suggests that eitherthe biogenesis and/or the cell surface docking and fusion potential of these vesicles is impaired in MPS IIIA. If this also occurs in central nervous system neurons, the reduction in neurotransmitter release could help to explain the developmentof neuropathology in MPS IIIA Refereed/Peer-reviewed
- Published
- 2012
7. Virtual work conditions impact negative work behaviors via ambiguity, anonymity, and (un)accountability: An integrative review.
- Author
-
Keating DJ, Cullen-Lester KL, and Meuser JD
- Subjects
- Humans, Workplace psychology, Social Responsibility, Crime Victims psychology
- Abstract
Negative work behavior (NWB) occurs with concerning frequency in virtual work environments. Despite their prevalence and a substantial, multidisciplinary research literature on virtual negative behaviors in general, we lack clear answers regarding if, how, and why conditions differentiating virtual (i.e., computer-mediated) from face-to-face (F2F) work impact perpetrators', victims', and bystanders' involvement in NWB. These questions remain because of an assumed isomorphism (i.e., identical form) within the literature on NWB in F2F and virtual work. We explain why we cannot assume that what is known about perpetrator engagement, victim experience, and bystander intervention from studying F2F NWB applies uniformly to virtual negative work behavior (VNWB). Specifically, we identify how eight conditions of the virtual workplace facilitate three psychological enablers (i.e., ambiguity, anonymity, and (un)accountability) of perpetrators', victims', and bystanders' involvement in VNWB. In doing so, this integrative conceptual review advances a coherent understanding of what is (un)known about VNWB, integrates fragmented theoretical literature, and guides practical intervention. Importantly, we identify limitations of existing research practices that threaten the validity and generalizability of empirical findings. If not addressed, these issues will continue to undermine theoretical development and empirical investigations of F2F NWB and VNWB. Finally, this review points to new areas of inquiry that will meaningfully advance the understanding of NWB in the modern, increasingly virtual workplace. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
- Published
- 2024
- Full Text
- View/download PDF
8. Alterations in GLP-1 and PYY release with aging and body mass in the human gut.
- Author
-
Jones LA, Sun EW, Lumsden AL, Thorpe DW, Peterson RA, De Fontgalland D, Sposato L, Rabbitt P, Hollington P, Wattchow DA, and Keating DJ
- Abstract
The lining of our intestinal surface contains an array of hormone-producing cells that are collectively our bodies' largest endocrine cell reservoir. These "enteroendocrine" (EE) cells reside amongst the billions of absorptive epithelial and other cell types that line our gastrointestinal tract and can sense and respond to the ever-changing internal environment in our gut. EE cells release an array of important signalling molecules that can act as hormones, including glucagon-like peptide (GLP-1) and peptide YY (PYY) which are co-secreted from L cells. While much is known about the effects of these hormones on metabolism, insulin secretion and food intake, less is understood about their secretion from human intestinal tissue. In this study we assess whether GLP-1 and PYY release differs across human small and large intestinal tissue locations within the gastrointestinal tract, and/or by sex, body weight and the age of an individual. We identify that the release of both hormones is greater in more distal regions of the human colon, but is not different between sexes. We observe a negative correlation of GLP-1 and BMI in the small, but not large, intestine. Increased aging correlates with declining secretion of both GLP-1 and PYY in human large, but not small, intestine. When the data for large intestine is isolated by region, this relationship with age remains significant for GLP-1 in the ascending and descending colon and in the descending colon for PYY. This is the first demonstration that site-specific differences in GLP-1 and PYY release occur in human gut, as do site-specific relationships of L cell secretion with aging and body mass., Competing Interests: Declaration of competing interest None., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2023
- Full Text
- View/download PDF
9. The Impact of Long-Term Macrolide Exposure on the Gut Microbiome and Its Implications for Metabolic Control.
- Author
-
Choo JM, Martin AM, Taylor SL, Sun E, Mobegi FM, Kanno T, Richard A, Burr LD, Lingman S, Martin M, Keating DJ, Mason AJ, and Rogers GB
- Subjects
- Animals, Mice, Macrolides pharmacology, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents therapeutic use, Erythromycin pharmacology, Gastrointestinal Microbiome, Respiratory Tract Diseases drug therapy
- Abstract
Long-term low-dose macrolide therapy is now widely used in the treatment of chronic respiratory diseases for its immune-modulating effects, although the antimicrobial properties of macrolides can also have collateral impacts on the gut microbiome. We investigated whether such treatment altered intestinal commensal microbiology and whether any such changes affected systemic immune and metabolic regulation. In healthy adults exposed to 4 weeks of low-dose erythromycin or azithromycin, as used clinically, we observed consistent shifts in gut microbiome composition, with a reduction in microbial capacity related to carbohydrate metabolism and short-chain fatty acid biosynthesis. These changes were accompanied by alterations in systemic biomarkers relating to immune (interleukin 5 [IL-5], IL-10, monocyte chemoattractant protein 1 [MCP-1]) and metabolic (serotonin [5-HT], C-peptide) homeostasis. Transplantation of erythromycin-exposed murine microbiota into germ-free mice demonstrated that changes in metabolic homeostasis and gastrointestinal motility, but not systemic immune regulation, resulted from changes in intestinal microbiology caused by macrolide treatment. Our findings highlight the potential for long-term low-dose macrolide therapy to influence host physiology via alteration of the gut microbiome. IMPORTANCE Long-term macrolide therapy is widely used in chronic respiratory diseases although its antibacterial activity can also affect the gut microbiota, a key regulator of host physiology. Macrolide-associated studies on the gut microbiota have been limited to short antibiotic courses and have not examined its consequences for host immune and metabolic regulation. This study revealed that long-term macrolides depleted keystone bacteria and impacted host regulation, mediated directly by macrolide activity or indirectly by alterations to the gut microbiota. Understanding these macrolide-associated mechanisms will contribute to identifying the risk of long-term exposure and highlights the importance of targeted therapy for maintenance of the gut microbiota., Competing Interests: The authors declare no conflict of interest.
- Published
- 2023
- Full Text
- View/download PDF
10. Single-cell gene expression links SARS-CoV-2 infection and gut serotonin.
- Author
-
Martin AM, Roach M, Jones LA, Thorpe D, Coleman RA, Allman C, Edwards R, and Keating DJ
- Subjects
- Humans, Serotonin, SARS-CoV-2, Gastrointestinal Tract, Gene Expression, COVID-19 genetics
- Abstract
Competing Interests: Competing interests: None declared.
- Published
- 2023
- Full Text
- View/download PDF
11. Distinguishing the contributions of neuronal and mucosal serotonin in the regulation of colonic motility.
- Author
-
Martin AM, Jones LA, Wei L, Spencer NJ, Sanders KM, Ro S, and Keating DJ
- Subjects
- Animals, Colon, Enterochromaffin Cells, Fluoxetine pharmacology, Intestinal Mucosa, Mice, Serotonergic Neurons, Gastrointestinal Motility physiology, Serotonin pharmacology
- Abstract
Background: Specialized enterochromaffin (EC) cells within the mucosal lining of the gut synthesize and secrete almost all serotonin (5-hydroxytryptamine, 5-HT) in the body. Significantly lower amounts of 5-HT are made by other peripheral tissues and serotonergic neurons within the enteric nervous system (ENS). EC cells are in close proximity to 5-HT receptors in the ENS, and the role of 5-HT as a modulator of gut motility, particularly colonic motor complexes, has been well defined. However, the relative contribution of neuronal 5-HT to this process under resting and stimulus-evoked conditions is unclear., Methods: In this study, we combined the use of the selective serotonin transporter (SERT) inhibitor, fluoxetine, with two models of mucosal 5-HT depletion-surgical removal of the mucosa and our Tph1
Cre/ERT2 ; Rosa26DTA mouse line-to determine the relative contribution of neuronal and mucosal 5-HT to resting and distension-evoked colonic motility., Key Results: Fluoxetine significantly reduced the frequency of colonic migrating complexes (CMCs) in flat-sheet preparations with the mucosa present and in intact control Tph1-DTA colons in which EC cells were present. No such effect was observed in mucosa-free preparations or in intact Tph1-DTA preparations lacking EC cell 5-HT., Conclusions and Inferences: We demonstrate that mucosal 5-HT release plays an important role in distension-evoked colonic motility, and that SERT inhibition no longer alters gut motility when EC cells are absent, thus demonstrating that ENS 5-HT does not play a role in regulating gut motility., (© 2022 John Wiley & Sons Ltd.)- Published
- 2022
- Full Text
- View/download PDF
12. Role of 5-HT in the enteric nervous system and enteroendocrine cells.
- Author
-
Spencer NJ and Keating DJ
- Abstract
Since the 1950s, considerable circumstantial evidence had been presented that endogenous 5-HT (serotonin) synthesized from within the wall of the gastrointestinal (GI) tract played an important role in GI motility and transit. However, identifying the precise functional role of gut-derived 5-HT has been difficult to ascertain, for a number of reasons. Over the past decade, as recording techniques have advanced significantly and access to new genetically modified animals improved, there have been major new insights and major changes in our understanding of the functional role of endogenous 5-HT in the GI tract. Data from many different laboratories have shown that major patterns of GI motility and transit still occur with minor or no, change when all endogenous 5-HT is pharmacologically or genetically ablated from the gut. Furthermore, antagonists of 5-HT
3 receptors are equally, or more potent at inhibiting GI motility in segments of intestine that are completely depleted of endogenous 5-HT. Here, the most recent findings are discussed with regard to the functional role of endogenous 5-HT in enterochromaffin cells and enteric neurons in gut motility and more broadly in some major homeostatic pathways., (© 2022 British Pharmacological Society.)- Published
- 2022
- Full Text
- View/download PDF
13. The gut-brain axis: spatial relationship between spinal afferent nerves and 5-HT-containing enterochromaffin cells in mucosa of mouse colon.
- Author
-
Dodds KN, Travis L, Kyloh MA, Jones LA, Keating DJ, and Spencer NJ
- Subjects
- Animals, Brain-Gut Axis, Colon metabolism, Mice, Enterochromaffin Cells metabolism, Serotonin metabolism
- Abstract
Cross talk between the gastrointestinal tract and brain is of significant relevance for human health and disease. However, our understanding of how the gut and brain communicate has been limited by a lack of techniques to identify the precise spatial relationship between extrinsic nerve endings and their proximity to specific cell types that line the inner surface of the gastrointestinal tract. We used an in vivo anterograde tracing technique, previously developed in our laboratory, to selectively label single spinal afferent axons and their nerve endings in mouse colonic mucosa. The closest three-dimensional distances between spinal afferent nerve endings and axonal varicosities to enterochromaffin (EC) cells, which contain serotonin (5-hydroxytryptamine; 5-HT), were then measured. The mean distances (± standard deviation) between any varicosity along a spinal afferent axon or its nerve ending, and the nearest EC cell, were 5.7 ± 6.0 μm (median: 3.6 μm) and 26.9 ± 18.6 μm (median: 24.1 μm), respectively. Randomization of the spatial location of EC cells revealed similar results to this actual data. These distances are ∼200-1,000 times greater than those between pre- and postsynaptic membranes (15-25 nm) that underlie synaptic transmission in the vertebrate nervous system. Our findings suggest that colonic 5-HT-containing EC cells release substances to activate centrally projecting spinal afferent nerves likely via diffusion, as such signaling is unlikely to occur with the spatial fidelity of a synapse. NEW & NOTEWORTHY We show an absence of close physical contact between spinal afferent nerves and 5-HT-containing EC cells in mouse colonic mucosa. Similar relative distances were observed between randomized EC cells and spinal afferents compared with actual data. This spatial relationship suggests that substances released from colonic 5-HT-containing EC cells are unlikely to act via synaptic transmission to neighboring spinal afferents that relay sensory information from the gut lumen to the brain.
- Published
- 2022
- Full Text
- View/download PDF
14. Diminished Piezo2-Dependent Tactile Sensitivity Occurs in Aging Human Gut and Slows Gastrointestinal Transit in Mice.
- Author
-
Jones LA, Jin B, Martin AM, Wei L, Ro S, and Keating DJ
- Subjects
- Aging, Animals, Digestion, Humans, Ion Channels metabolism, Mechanotransduction, Cellular, Mice, Gastrointestinal Transit, Touch
- Published
- 2022
- Full Text
- View/download PDF
15. The gut microbiome and mental health: advances in research and emerging priorities.
- Author
-
Shoubridge AP, Choo JM, Martin AM, Keating DJ, Wong ML, Licinio J, and Rogers GB
- Subjects
- Brain microbiology, Humans, Mental Health, Gastrointestinal Microbiome physiology, Mental Disorders microbiology, Microbiota physiology
- Abstract
The gut microbiome exerts a considerable influence on human neurophysiology and mental health. Interactions between intestinal microbiology and host regulatory systems have now been implicated both in the development of psychiatric conditions and in the efficacy of many common therapies. With the growing acceptance of the role played by the gut microbiome in mental health outcomes, the focus of research is now beginning to shift from identifying relationships between intestinal microbiology and pathophysiology, and towards using this newfound insight to improve clinical outcomes. Here, we review recent advances in our understanding of gut microbiome-brain interactions, the mechanistic underpinnings of these relationships, and the ongoing challenge of distinguishing association and causation. We set out an overarching model of the evolution of microbiome-CNS interaction and examine how a growing knowledge of these complex systems can be used to determine disease susceptibility and reduce risk in a targeted manner., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)
- Published
- 2022
- Full Text
- View/download PDF
16. Pharmacological and structure-activity relationship studies of oleoyl-lysophosphatidylinositol synthetic mimetics.
- Author
-
Paternoster S, Simpson PV, Kokh E, Kizilkaya HS, Rosenkilde MM, Mancera RL, Keating DJ, Massi M, and Falasca M
- Subjects
- Animals, Cell Line, Enteroendocrine Cells metabolism, Humans, Lysophospholipids chemistry, Mice, Inbred C57BL, Models, Molecular, Receptors, G-Protein-Coupled metabolism, Structure-Activity Relationship, Mice, Diabetes Mellitus, Experimental metabolism, Enteroendocrine Cells drug effects, Glucagon-Like Peptide 1 metabolism, Lysophospholipids pharmacology
- Abstract
Metabolic diseases, such as obesity and type 2 diabetes, are relentlessly spreading worldwide. The beginning of the 21st century has seen the introduction of mechanistically novel types of drugs, aimed primarily at keeping these pathologies under control. In particular, an important family of therapeutics exploits the beneficial physiology of the gut-derived glucagon-like peptide-1 (GLP-1), with important clinical benefits, from glycaemic control to cardioprotection. Nonetheless, these protein-based drugs act systemically as exogenous GLP-1 mimetics and are not exempt from side effects. The food-derived lipid oleoyl-lysophosphatidylinositol (LPI) is a potent GPR119-dependent GLP-1 secreting agent. Here we present a structure-activity relationship (SAR) study of a synthetic library of oleoyl-LPI mimetics capable to induce the physiological release of GLP-1 from gastrointestinal enteroendocrine cells (EECs). The best lead compounds have shown potent and efficient release of GLP-1 in vitro from human and murine cells, and in vivo in diabetic db/db mice. We have also generated a molecular model of oleoyl-LPI, as well as its best performing analogues, interacting with the orthosteric site of GPR119, laying foundational evidence for their pharmacological activity., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
17. Dynamin regulates L cell secretion in human gut.
- Author
-
Sun EW, Matusica D, Wattchow DA, McCluskey A, Robinson PJ, and Keating DJ
- Subjects
- Adult, Aged, Animals, Cells, Cultured, Culture Media chemistry, Cyanoacrylates pharmacology, Enteroendocrine Cells drug effects, Enteroendocrine Cells metabolism, Female, Humans, Indoles pharmacology, Intestinal Mucosa drug effects, Intestinal Mucosa metabolism, L Cells, Male, Mice, Middle Aged, Tyrphostins pharmacology, Dynamin I metabolism, Dynamin II metabolism, Enteroendocrine Cells cytology, Glucagon-Like Peptide 1 metabolism, Intestinal Mucosa cytology, Peptide YY metabolism
- Abstract
Background: The mechanochemical enzyme dynamin mediates endocytosis and regulates neuroendocrine cell exocytosis. Enteroendocrine L cells co-secrete the anorectic gut hormones glucagon-like peptide 1 (GLP-1) and peptide YY (PYY) postprandially and is a potential therapeutic target for metabolic diseases. In the present study, we aimed to determine if dynamin is implicated in human L cell secretion., Methods: Western blot was performed on the murine L cell line GLUTag. Static incubation of human colonic mucosae with activators and inhibitors of dynamin was carried out. GLP-1 and PYY contents of the secretion supernatants were assayed using ELISA., Results and Conclusion: s: Both dynamin I and II are expressed in GLUTag cells. The dynamin activator Ryngo 1-23 evoked significant GLP-1 and PYY release from human colonic mucosae while the dynamin inhibitor Dynole 3-42 significantly inhibited release triggered by known L cell secretagogues. Thus, the cell signaling regulator dynamin is able to bi-directionally regulate L cell hormone secretion in the human gut and may represent a novel target for gastrointestinal-targeted metabolic drug development., (Copyright © 2021. Published by Elsevier B.V.)
- Published
- 2021
- Full Text
- View/download PDF
18. The composition of the gut microbiota following early-life antibiotic exposure affects host health and longevity in later life.
- Author
-
Lynn MA, Eden G, Ryan FJ, Bensalem J, Wang X, Blake SJ, Choo JM, Chern YT, Sribnaia A, James J, Benson SC, Sandeman L, Xie J, Hassiotis S, Sun EW, Martin AM, Keller MD, Keating DJ, Sargeant TJ, Proud CG, Wesselingh SL, Rogers GB, and Lynn DJ
- Subjects
- Animals, Longevity drug effects, Mice, Anti-Bacterial Agents pharmacology, Gastrointestinal Microbiome drug effects, Gastrointestinal Microbiome immunology, Longevity immunology
- Abstract
Studies investigating whether there is a causative link between the gut microbiota and lifespan have largely been restricted to invertebrates or to mice with a reduced lifespan because of a genetic deficiency. We investigate the effect of early-life antibiotic exposure on otherwise healthy, normal chow-fed, wild-type mice, monitoring these mice for more than 700 days in comparison with untreated control mice. We demonstrate the emergence of two different low-diversity community types, post-antibiotic microbiota (PAM) I and PAM II, following antibiotic exposure. PAM II but not PAM I mice have impaired immunity, increased insulin resistance, and evidence of increased inflammaging in later life as well as a reduced lifespan. Our data suggest that differences in the composition of the gut microbiota following antibiotic exposure differentially affect host health and longevity in later life., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
19. A Gut-Intrinsic Melanocortin Signaling Complex Augments L-Cell Secretion in Humans.
- Author
-
Sun EW, Iepsen EW, Pezos N, Lumsden AL, Martin AM, Schober G, Isaacs NJ, Rayner CK, Nguyen NQ, de Fontgalland D, Rabbitt P, Hollington P, Wattchow DA, Hansen T, Holm JC, Liou AP, Jackson VM, Torekov SS, Young RL, and Keating DJ
- Subjects
- Autocrine Communication, Blood Glucose metabolism, Case-Control Studies, Enteroendocrine Cells drug effects, Glucose administration & dosage, Glucose Tolerance Test, Humans, Intestinal Mucosa drug effects, Loss of Function Mutation, Paracrine Communication, Pro-Opiomelanocortin genetics, Receptor, Melanocortin, Type 4 agonists, Receptor, Melanocortin, Type 4 genetics, Secretory Pathway, Signal Transduction, Time Factors, alpha-MSH pharmacology, Enteroendocrine Cells metabolism, Glucagon-Like Peptide 1 metabolism, Intestinal Mucosa metabolism, Peptide YY metabolism, Pro-Opiomelanocortin metabolism, Receptor, Melanocortin, Type 4 metabolism, alpha-MSH metabolism
- Abstract
Objective: Hypothalamic melanocortin 4 receptors (MC4R) are a key regulator of energy homeostasis. Brain-penetrant MC4R agonists have failed, as concentrations required to suppress food intake also increase blood pressure. However, peripherally located MC4R may also mediate metabolic benefits of MC4R activation. Mc4r transcript is enriched in mouse enteroendocrine L cells and peripheral administration of the endogenous MC4R agonist, α-melanocyte stimulating hormone (α-MSH), triggers the release of the anorectic hormones Glucagon-like peptide-1 (GLP-1) and peptide tyrosine tyrosine (PYY) in mice. This study aimed to determine whether pathways linking MC4R and L-cell secretion exist in humans., Design: GLP-1 and PYY levels were assessed in body mass index-matched individuals with or without loss-of-function MC4R mutations following an oral glucose tolerance test. Immunohistochemistry was performed on human intestinal sections to characterize the mucosal MC4R system. Static incubations with MC4R agonists were carried out on human intestinal epithelia, GLP-1 and PYY contents of secretion supernatants were assayed., Results: Fasting PYY levels and oral glucose-induced GLP-1 secretion were reduced in humans carrying a total loss-of-function MC4R mutation. MC4R was localized to L cells and regulates GLP-1 and PYY secretion from ex vivo human intestine. α-MSH immunoreactivity in the human intestinal epithelia was predominantly localized to L cells. Glucose-sensitive mucosal pro-opiomelanocortin cells provide a local source of α-MSH that is essential for glucose-induced GLP-1 secretion in small intestine., Conclusion: Our findings describe a previously unidentified signaling nexus in the human gastrointestinal tract involving α-MSH release and MC4R activation on L cells in an autocrine and paracrine fashion. Outcomes from this study have direct implications for targeting mucosal MC4R to treat human metabolic disorders., (Crown Copyright © 2021. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
20. Serotonin Deficiency Is Associated With Delayed Gastric Emptying.
- Author
-
Wei L, Singh R, Ha SE, Martin AM, Jones LA, Jin B, Jorgensen BG, Zogg H, Chervo T, Gottfried-Blackmore A, Nguyen L, Habtezion A, Spencer NJ, Keating DJ, Sanders KM, and Ro S
- Subjects
- Animals, Cell Line, Enterochromaffin Cells physiology, Humans, Mice, Mice, Transgenic, Tryptophan Hydroxylase metabolism, Gastric Emptying genetics, Gastrointestinal Transit genetics, Serotonin deficiency
- Abstract
Background & Aims: Gastrointestinal (GI) motility is regulated by serotonin (5-hydroxytryptamine [5-HT]), which is primarily produced by enterochromaffin (EC) cells in the GI tract. However, the precise roles of EC cell-derived 5-HT in regulating gastric motility remain a major point of conjecture. Using a novel transgenic mouse line, we investigated the distribution of EC cells and the pathophysiologic roles of 5-HT deficiency in gastric motility in mice and humans., Methods: We developed an inducible, EC cell-specific Tph1
CreERT2/+ mouse, which was used to generate a reporter mouse line, Tph1-tdTom, and an EC cell-depleted line, Tph1-DTA. We examined EC cell distribution, morphology, and subpopulations in reporter mice. GI motility was measured in vivo and ex vivo in EC cell-depleted mice. Additionally, we evaluated 5-HT content in biopsy and plasma specimens from patients with idiopathic gastroparesis (IG)., Results: Tph1-tdTom mice showed EC cells that were heterogeneously distributed throughout the GI tract with the greatest abundance in the antrum and proximal colon. Two subpopulations of EC cells were identified in the gut: self-renewal cells located at the base of the crypt and mature cells observed in the villi. Tph1-DTA mice displayed delayed gastric emptying, total GI transit, and colonic transit. These gut motility alterations were reversed by exogenous provision of 5-HT. Patients with IG had a significant reduction of antral EC cell numbers and 5-HT content, which negatively correlated with gastric emptying rate., Conclusions: The Tph1CreERT2/+ mouse provides a powerful tool to study the functional roles of EC cells in the GI tract. Our findings suggest a new pathophysiologic mechanism of 5-HT deficiency in IG., (Copyright © 2021 AGA Institute. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
- Full Text
- View/download PDF
21. Evidence for Glucagon Secretion and Function Within the Human Gut.
- Author
-
Sun EW, Martin AM, de Fontgalland D, Sposato L, Rabbitt P, Hollington P, Wattchow DA, Colella AD, Chataway T, Wewer Albrechtsen NJ, Spencer NJ, Young RL, and Keating DJ
- Subjects
- Adult, Aged, Aged, 80 and over, Cholesterol metabolism, Cohort Studies, Female, Glucagon-Like Peptide 1 metabolism, Glucose metabolism, Humans, Male, Middle Aged, Glucagon metabolism, Intestinal Mucosa metabolism
- Abstract
Glucagon is secreted by pancreatic α cells in response to hypoglycemia and increases hepatic glucose output through hepatic glucagon receptors (GCGRs). There is evidence supporting the notion of extrapancreatic glucagon but its source and physiological functions remain elusive. Intestinal tissue samples were obtained from patients undergoing surgical resection of cancer. Mass spectrometry analysis was used to detect glucagon from mucosal lysate. Static incubations of mucosal tissue were performed to assess glucagon secretory response. Glucagon concentration was quantitated using a highly specific sandwich enzyme-linked immunosorbent assay. A cholesterol uptake assay and an isolated murine colonic motility assay were used to assess the physiological functions of intestinal GCGRs. Fully processed glucagon was detected by mass spectrometry in human intestinal mucosal lysate. High glucose evoked significant glucagon secretion from human ileal tissue independent of sodium glucose cotransporter and KATP channels, contrasting glucose-induced glucagon-like peptide 1 (GLP-1) secretion. The GLP-1 receptor agonist Exendin-4 attenuated glucose-induced glucagon secretion from the human ileum. GCGR blockade significantly increased cholesterol uptake in human ileal crypt culture and markedly slowed ex vivo colonic motility. Our findings describe the human gut as a potential source of extrapancreatic glucagon and demonstrate a novel enteric glucagon/GCGR circuit with important physiological functions beyond glycemic regulation., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2021
- Full Text
- View/download PDF
22. Enteroendocrine cells sense bacterial tryptophan catabolites to activate enteric and vagal neuronal pathways.
- Author
-
Ye L, Bae M, Cassilly CD, Jabba SV, Thorpe DW, Martin AM, Lu HY, Wang J, Thompson JD, Lickwar CR, Poss KD, Keating DJ, Jordt SE, Clardy J, Liddle RA, and Rawls JF
- Subjects
- Animals, Animals, Genetically Modified, Cholinergic Neurons metabolism, Enteric Nervous System cytology, Gastrointestinal Motility physiology, Intestinal Mucosa cytology, Intestinal Mucosa innervation, Proto-Oncogene Proteins c-ret genetics, Serotonin metabolism, Signal Transduction, Tryptophan metabolism, Zebrafish, Zebrafish Proteins genetics, Edwardsiella tarda metabolism, Enteric Nervous System metabolism, Enteroendocrine Cells physiology, Intestinal Mucosa metabolism, TRPA1 Cation Channel metabolism
- Abstract
The intestinal epithelium senses nutritional and microbial stimuli using epithelial sensory enteroendocrine cells (EEC). EECs communicate nutritional information to the nervous system, but whether they also relay signals from intestinal microbes remains unknown. Using in vivo real-time measurements of EEC and nervous system activity in zebrafish, we discovered that the bacteria Edwardsiella tarda activate EECs through the receptor transient receptor potential ankyrin A1 (Trpa1) and increase intestinal motility. Microbial, pharmacological, or optogenetic activation of Trpa1
+ EECs directly stimulates vagal sensory ganglia and activates cholinergic enteric neurons by secreting the neurotransmitter 5-hydroxytryptamine (5-HT). A subset of indole derivatives of tryptophan catabolism produced by E. tarda and other gut microbes activates zebrafish EEC Trpa1 signaling. These catabolites also directly stimulate human and mouse Trpa1 and intestinal 5-HT secretion. These results establish a molecular pathway by which EECs regulate enteric and vagal neuronal pathways in response to microbial signals., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)- Published
- 2021
- Full Text
- View/download PDF
23. Circulating cathepsin S improves glycaemic control in mice.
- Author
-
Karimkhanloo H, Keenan SN, Sun EW, Wattchow DA, Keating DJ, Montgomery MK, and Watt MJ
- Subjects
- 3T3-L1 Cells, Adipocytes metabolism, Animals, Cathepsins therapeutic use, Diabetes Mellitus, Type 2 drug therapy, Drug Evaluation, Preclinical, Glucagon-Like Peptide 1 metabolism, Glucose metabolism, Glycemic Control, Liver metabolism, Mice, Blood Glucose, Cathepsins blood
- Abstract
Cathepsin S (CTSS) is a cysteine protease that regulates many physiological processes and is increased in obesity and type 2 diabetes. While previous studies show that deletion of CTSS improves glycaemic control through suppression of hepatic glucose output, little is known about the role of circulating CTSS in regulating glucose and energy metabolism. We assessed the effects of recombinant CTSS on metabolism in cultured hepatocytes, myotubes and adipocytes, and in mice following acute CTSS administration. CTSS improved glucose tolerance in lean mice and this coincided with increased plasma insulin. CTSS reduced G6pc and Pck1 mRNA expression and glucose output from hepatocytes but did not affect glucose metabolism in myotubes or adipocytes. CTSS did not affect insulin secretion from pancreatic β-cells, rather CTSS stimulated glucagon-like peptide (GLP)-1 secretion from intestinal mucosal tissues. CTSS retained its positive effects on glycaemic control in mice injected with the GLP1 receptor antagonist Exendin (9-39) amide. The effects of CTSS on glycaemic control were not retained in high-fat-fed mice or db/db mice, despite the preservation of CTSS' inhibitory actions on hepatic glucose output in isolated primary hepatocytes. In conclusion, we unveil a role for CTSS in the regulation of glycaemic control via direct effects on hepatocytes, and that these effects on glycaemic control are abrogated in insulin resistant states.
- Published
- 2021
- Full Text
- View/download PDF
24. Amperometry in Single Cells and Tissue.
- Author
-
Keating DJ
- Subjects
- Endocrine System metabolism, Humans, Kinetics, Nervous System metabolism, Endocrine System ultrastructure, Exocytosis genetics, Nervous System ultrastructure, Single-Cell Analysis methods
- Abstract
The release from cells of signaling molecules through the controlled process of exocytosis involves multiple coordinated steps and is essential for the proper control of a multitude of biological pathways across the endocrine and nervous systems. However, these events are minute both temporally and in terms of the minute amounts of neurotransmitters, hormones, growth factors, and peptides released from single vesicles during exocytosis. It is therefore difficult to measure the kinetics of single exocytosis events in real time. One noninvasive method of measuring the release of molecules from cells is carbon-fiber amperometry. In this chapter, we will describe how we undertake such measurements from both single cells and in live tissue, how the subsequent data are analyzed, and how we interpret these results in terms of their relevant physiology.
- Published
- 2021
- Full Text
- View/download PDF
25. The Suitability of Glioblastoma Cell Lines as Models for Primary Glioblastoma Cell Metabolism.
- Author
-
Arthurs AL, Keating DJ, Stringer BW, and Conn SJ
- Abstract
In contrast to most non-malignant tissue, cells comprising the brain tumour glioblastoma (GBM) preferentially utilise glycolysis for metabolism via "the Warburg effect". Research into therapeutics targeting the disease's highly glycolytic state offer a promising avenue to improve patient survival. These studies often employ GBM cell lines for in vitro studies which translate poorly to the in vivo patient context. The metabolic traits of five of the most used GBM cell lines were assessed and compared to primary GBM and matched, healthy brain tissue. In patient-derived GBM cell lines, the basal mitochondrial rate ( p = 0.043) and ATP-linked respiration ( p < 0.001) were lower than primary adjacent normal cells from the same patient, while reserve capacity ( p = 0.037) and Krebs cycle capacity ( p = 0.002) were higher. Three cell lines, U251MG, U373MG and D54, replicate the mitochondrial metabolism of primary GBM cells. Surprisingly, glycolytic capacity is not different between healthy and GBM tissue. The T98G cell line recapitulated glycolysis-related metabolic parameters of the primary GBM cells and is recommended for research relating to glycolysis. These findings can guide preclinical research into the development of novel therapeutics targeting metabolic pathways in GBM.
- Published
- 2020
- Full Text
- View/download PDF
26. The ever-changing roles of serotonin.
- Author
-
Jones LA, Sun EW, Martin AM, and Keating DJ
- Subjects
- Adipocytes enzymology, Adipocytes metabolism, Animals, Enteric Nervous System metabolism, Enterochromaffin Cells enzymology, Gastrointestinal Motility physiology, Gastrointestinal Tract enzymology, Gastrointestinal Tract microbiology, Homeostasis, Humans, Neurons enzymology, Tryptophan Hydroxylase metabolism, Enterochromaffin Cells metabolism, Gastrointestinal Microbiome physiology, Gastrointestinal Tract metabolism, Glucose metabolism, Neurons metabolism, Obesity metabolism, Serotonin metabolism
- Abstract
Serotonin (5-HT) has traditional roles as a key neurotransmitter in the central nervous system and as a regulatory hormone controlling a broad range of physiological functions. Perhaps the most classically-defined functions of 5-HT are centrally in the control of mood, sleep and anxiety and peripherally in the modulation of gastrointestinal motility. A more recently appreciated role for 5-HT has emerged, however, as an important metabolic hormone contributing to glucose homeostasis and adiposity, with a causal relationship existing between circulating 5-HT levels and metabolic diseases. Almost all peripheral 5-HT is derived from specialised enteroendocrine cells, called enterochromaffin (EC) cells, located throughout the length of the lining of the gastrointestinal tract. EC cells are important luminal sensory cells that can detect and respond to an array of ingested nutrients, as well as luminal gut microbiota and their associated metabolites. Intriguingly, the interaction between gut microbiota and EC cells is dynamic in nature and has strong implications for host physiology. In this review, we discuss the traditional and modern functions of 5-HT and highlight an emerging pathway by which gut microbiota influences host health. Serotonin, also known as 5-hydroxytryptamine (5-HT), is an important neurotransmitter, growth factor and hormone that mediates a range of physiological functions. In mammals, serotonin is synthesized from the essential amino acid tryptophan by the rate-limiting enzyme tryptophan hydroxylase (TPH), for which there are two isoforms expressed in distinct cell types throughout the body. Tph1 is mainly expressed by specialized gut endocrine cells known as enterochromaffin (EC) cells and by other non-neuronal cell types such as adipocytes (Walther et al., 2003). Tph2 is primarily expressed in neurons of the raphe nuclei of the brain stem and a subset of neurons in the enteric nervous system (ENS) (Yabut et al., 2019). As 5-HT cannot readily cross the blood-brain barrier, the central and peripheral pools of 5-HT are anatomically separated and as such, act in their own distinct manners (Martin et al., 2017c). In this review we discuss the peripheral roles of serotonin, with particular focus on the interaction of gut-derived serotonin with the gut microbiota, and address emerging evidence linking this relationship with host homeostasis., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
27. Diet differentially regulates enterochromaffin cell serotonin content, density and nutrient sensitivity in the mouse small and large intestine.
- Author
-
Martin AM, Jones LA, Jessup CF, Sun EW, and Keating DJ
- Subjects
- Animals, Blood Glucose metabolism, Male, Mice, Diet, Enterochromaffin Cells metabolism, Intestine, Large metabolism, Intestine, Small metabolism, Obesity metabolism, Serotonin metabolism
- Abstract
Background: Enterochromaffin (EC) cells are specialized enteroendocrine cells lining the gastrointestinal (GI) tract and the source of almost all serotonin (5-hydroxytryptamine; 5-HT) in the body. Gut-derived 5-HT has a plethora of physiological roles, including regulation of gastrointestinal motility, and has been implicated as a driver of obesity and metabolic disease. This is due to 5-HT influencing key metabolic processes, such as hepatic gluconeogenesis, adipose tissue lipolysis and hindering thermogenic capacity. Increased circulating 5-HT occurs in humans with obesity and type 2 diabetes. However, despite the known metabolic roles of gut-derived 5-HT, the mechanisms underlying the cellular-level change in EC cells under obesogenic conditions remains unknown., Methods: We use a mouse model of diet-induced obesity (DIO) to identify the regional changes that occur in primary EC cells from the duodenum and colon. Transcriptional changes in the nutrient sensing profile of primary EC cells were assessed, and responses to nutrient stimuli in culture were determined by 5-HT ELISA., Key Results: We find that obesogenic conditions affect EC cells in a region-dependent manner. Duodenal EC cells from DIO mice have impaired sugar sensing even in the presence of increased 5-HT content per cell, while colonic EC cell numbers are significantly increased, but have unaltered nutrient sensing capacity., Conclusions & Inferences: Our findings from this study add novel insights into the mechanisms by which functional changes to EC cells occur at a cellular level, which may contribute to the altered circulating 5-HT seen with obesity and metabolic disease, and associated gastrointestinal disorders., (© 2020 John Wiley & Sons Ltd.)
- Published
- 2020
- Full Text
- View/download PDF
28. Islets and pancreatic ductal adenocarcinoma - An opportunity for translational research from the 'Bench to the Bedside'.
- Author
-
Barreto SG, Michael MZ, and Keating DJ
- Subjects
- Adenocarcinoma pathology, Adenoma, Islet Cell pathology, Carcinoma, Pancreatic Ductal pathology, Humans, Pancreatic Neoplasms pathology, Translational Research, Biomedical, Adenocarcinoma therapy, Adenoma, Islet Cell therapy, Carcinoma, Pancreatic Ductal therapy, Islets of Langerhans pathology, Pancreatic Neoplasms therapy
- Abstract
The islet-acinar axis is of prime importance to the optimal functioning of the human pancreas. Not only is this inter-relationship important for normal physiological processes, it is also relevant in diseased states, including chronic pancreatitis and pancreatic ductal adenocarcinoma (PDAC). Early experiments, nearly 4 decades ago, explored the role of islets in the development and progression of PDAC. These led to further studies that provided compelling evidence to support the role of islets and their hormones in PDAC. This association presents oncologists with therapeutic options not only for managing, but potentially preventing PDAC, a cancer that is well known for its poor patient outcomes. This review will discuss the accumulated evidence regarding the role of islets and their hormones in PDAC and highlight areas for future research., Competing Interests: Declaration of competing interest None to declare., (Copyright © 2020 IAP and EPC. Published by Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
29. Mechanisms controlling hormone secretion in human gut and its relevance to metabolism.
- Author
-
Martin AM, Sun EW, and Keating DJ
- Subjects
- Cholecystokinin metabolism, Cytokines metabolism, Energy Metabolism, Enteroendocrine Cells metabolism, Gastrointestinal Microbiome, Glucagon-Like Peptide 1 metabolism, Glucose metabolism, Homeostasis, Humans, Peptide YY metabolism, Serotonin metabolism, Endocrine System metabolism, Gastrointestinal Hormones metabolism, Gastrointestinal Tract metabolism
- Abstract
The homoeostatic regulation of metabolism is highly complex and involves multiple inputs from both the nervous and endocrine systems. The gut is the largest endocrine organ in our body and synthesises and secretes over 20 different hormones from enteroendocrine cells that are dispersed throughout the gut epithelium. These hormones include GLP-1, PYY, GIP, serotonin, and CCK, each of whom play pivotal roles in maintaining energy balance and glucose homeostasis. Some are now the basis of several clinically used glucose-lowering and weight loss therapies. The environment in which these enteroendocrine cells exist is also complex, as they are exposed to numerous physiological inputs including ingested nutrients, circulating factors and metabolites produced from neighbouring gut microbiome. In this review, we examine the diverse means by which gut-derived hormones carry out their metabolic functions through their interactions with different metabolically important organs including the liver, pancreas, adipose tissue and brain. Furthermore, we discuss how nutrients and microbial metabolites affect gut hormone secretion and the mechanisms underlying these interactions.
- Published
- 2019
- Full Text
- View/download PDF
30. The gut microbiome regulates host glucose homeostasis via peripheral serotonin.
- Author
-
Martin AM, Yabut JM, Choo JM, Page AJ, Sun EW, Jessup CF, Wesselingh SL, Khan WI, Rogers GB, Steinberg GR, and Keating DJ
- Subjects
- Animals, Anti-Bacterial Agents pharmacology, Area Under Curve, Blood Glucose metabolism, Gene Deletion, Glucose Tolerance Test, Male, Mice, Mice, Inbred C57BL, Random Allocation, Gastrointestinal Microbiome, Glucose metabolism, Homeostasis, Serotonin physiology
- Abstract
The gut microbiome is an established regulator of aspects of host metabolism, such as glucose handling. Despite the known impacts of the gut microbiota on host glucose homeostasis, the underlying mechanisms are unknown. The gut microbiome is also a potent mediator of gut-derived serotonin synthesis, and this peripheral source of serotonin is itself a regulator of glucose homeostasis. Here, we determined whether the gut microbiome influences glucose homeostasis through effects on gut-derived serotonin. Using both pharmacological inhibition and genetic deletion of gut-derived serotonin synthesis, we find that the improvements in host glucose handling caused by antibiotic-induced changes in microbiota composition are dependent on the synthesis of peripheral serotonin., Competing Interests: The authors declare no conflict of interest., (Copyright © 2019 the Author(s). Published by PNAS.)
- Published
- 2019
- Full Text
- View/download PDF
31. Treatment of type 2 diabetes with the designer cytokine IC7Fc.
- Author
-
Findeisen M, Allen TL, Henstridge DC, Kammoun H, Brandon AE, Baggio LL, Watt KI, Pal M, Cron L, Estevez E, Yang C, Kowalski GM, O'Reilly L, Egan C, Sun E, Thai LM, Krippner G, Adams TE, Lee RS, Grötzinger J, Garbers C, Risis S, Kraakman MJ, Mellet NA, Sligar J, Kimber ET, Young RL, Cowley MA, Bruce CR, Meikle PJ, Baldock PA, Gregorevic P, Biden TJ, Cooney GJ, Keating DJ, Drucker DJ, Rose-John S, and Febbraio MA
- Subjects
- Adaptor Proteins, Signal Transducing metabolism, Animals, Binding, Competitive, Cytokines chemistry, Diabetes Mellitus, Type 2 metabolism, Drug Design, Fatty Liver prevention & control, Glucose Tolerance Test, Humans, Hyperglycemia drug therapy, Hyperglycemia metabolism, Incretins metabolism, Interleukin-6 antagonists & inhibitors, Interleukin-6 metabolism, Male, Mice, Muscle, Skeletal drug effects, Obesity metabolism, Pancreas metabolism, Phosphoproteins metabolism, Protein Engineering, Receptors, Interleukin-6 metabolism, Signal Transduction, Transcription Factors, Weight Gain drug effects, YAP-Signaling Proteins, Cytokine Receptor gp130 metabolism, Cytokines chemical synthesis, Cytokines therapeutic use, Diabetes Mellitus, Type 2 drug therapy, Immunoglobulin G therapeutic use, Recombinant Fusion Proteins therapeutic use
- Abstract
The gp130 receptor cytokines IL-6 and CNTF improve metabolic homeostasis but have limited therapeutic use for the treatment of type 2 diabetes. Accordingly, we engineered the gp130 ligand IC7Fc, in which one gp130-binding site is removed from IL-6 and replaced with the LIF-receptor-binding site from CNTF, fused with the Fc domain of immunoglobulin G, creating a cytokine with CNTF-like, but IL-6-receptor-dependent, signalling. Here we show that IC7Fc improves glucose tolerance and hyperglycaemia and prevents weight gain and liver steatosis in mice. In addition, IC7Fc either increases, or prevents the loss of, skeletal muscle mass by activation of the transcriptional regulator YAP1. In human-cell-based assays, and in non-human primates, IC7Fc treatment results in no signs of inflammation or immunogenicity. Thus, IC7Fc is a realistic next-generation biological agent for the treatment of type 2 diabetes and muscle atrophy, disorders that are currently pandemic.
- Published
- 2019
- Full Text
- View/download PDF
32. Emerging Roles for Serotonin in Regulating Metabolism: New Implications for an Ancient Molecule.
- Author
-
Yabut JM, Crane JD, Green AE, Keating DJ, Khan WI, and Steinberg GR
- Subjects
- Adipose Tissue, Brown metabolism, Adipose Tissue, White metabolism, Animals, Energy Metabolism, Humans, Lipid Metabolism, Serotonin physiology, Signal Transduction, Serotonin metabolism
- Abstract
Serotonin is a phylogenetically ancient biogenic amine that has played an integral role in maintaining energy homeostasis for billions of years. In mammals, serotonin produced within the central nervous system regulates behavior, suppresses appetite, and promotes energy expenditure by increasing sympathetic drive to brown adipose tissue. In addition to these central circuits, emerging evidence also suggests an important role for peripheral serotonin as a factor that enhances nutrient absorption and storage. Specifically, glucose and fatty acids stimulate the release of serotonin from the duodenum, promoting gut peristalsis and nutrient absorption. Serotonin also enters the bloodstream and interacts with multiple organs, priming the body for energy storage by promoting insulin secretion and de novo lipogenesis in the liver and white adipose tissue, while reducing lipolysis and the metabolic activity of brown and beige adipose tissue. Collectively, peripheral serotonin acts as an endocrine factor to promote the efficient storage of energy by upregulating lipid anabolism. Pharmacological inhibition of serotonin synthesis or signaling in key metabolic tissues are potential drug targets for obesity, type 2 diabetes, and nonalcoholic fatty liver disease (NAFLD)., (Copyright © 2019 Endocrine Society.)
- Published
- 2019
- Full Text
- View/download PDF
33. Metformin Triggers PYY Secretion in Human Gut Mucosa.
- Author
-
Sun EW, Martin AM, Wattchow DA, de Fontgalland D, Rabbitt P, Hollington P, Young RL, and Keating DJ
- Subjects
- AMP-Activated Protein Kinases antagonists & inhibitors, AMP-Activated Protein Kinases metabolism, Adult, Aged, Colon cytology, Colon drug effects, Colon metabolism, Diabetes Mellitus, Type 2 drug therapy, Diabetes Mellitus, Type 2 metabolism, Enteroendocrine Cells drug effects, Enteroendocrine Cells metabolism, Equilibrative Nucleoside Transport Proteins metabolism, Female, Humans, Hypoglycemic Agents therapeutic use, Ileum cytology, Ileum drug effects, Ileum metabolism, Intestinal Mucosa cytology, Intestinal Mucosa metabolism, Male, Metformin therapeutic use, Middle Aged, Serotonin Plasma Membrane Transport Proteins metabolism, Weight Loss drug effects, Hypoglycemic Agents pharmacology, Intestinal Mucosa drug effects, Metformin pharmacology, Peptide YY metabolism
- Abstract
Context: The antidiabetic drug metformin causes weight loss, but the underlying mechanisms are unclear. Recent clinical studies show that metformin increases plasma levels of the anorectic gut hormone, peptide YY (PYY), but whether this is through a direct effect on the gut is unknown., Objective: We hypothesized that exposure of human gut mucosal tissue to metformin would acutely trigger PYY secretion., Design, Setting, Participants, and Interventions: Mucosal tissue was prepared from 46 human colonic and 9 ileal samples obtained after surgical resection and ex vivo secretion assays were performed. Tissue was exposed to metformin, as well as a series of other compounds as part of our mechanistic studies, in static incubations. Supernatant was sampled after 15 minutes., Main Outcome Measures: PYY levels in supernatant, measured using ELISA., Results: Metformin increased PYY secretion from both ileal (P < 0.05) and colonic (P < 0.001) epithelia. Both basal and metformin-induced PYY secretion were unchanged across body mass index or in tissues obtained from individuals with type 2 diabetes. Metformin-dependent PYY secretion was blocked by inhibitors of the plasma membrane monoamine transporter (PMAT) and the serotonin reuptake transporter (SERT), as well as by an inhibitor of AMP kinase (AMPK)., Conclusions: This is a report of a direct action of metformin on the gut epithelium to trigger PYY secretion in humans, occurring via cell internalization through PMAT and SERT and intracellular activation of AMPK. Our results provide further support that the role of metformin in the treatment of metabolic syndrome has a gut-based component., (Copyright © 2019 Endocrine Society.)
- Published
- 2019
- Full Text
- View/download PDF
34. Extracellular and intracellular sphingosine-1-phosphate distinctly regulates exocytosis in chromaffin cells.
- Author
-
Jiang ZJ, Delaney TL, Zanin MP, Haberberger RV, Pitson SM, Huang J, Alford S, Cologna SM, Keating DJ, and Gong LW
- Subjects
- Animals, Female, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Sphingosine metabolism, Chromaffin Cells metabolism, Exocytosis physiology, Lysophospholipids metabolism, Sphingosine analogs & derivatives
- Abstract
Sphingosine-1-phosphate (S1P) is an essential bioactive sphingosine lipid involved in many neurological disorders. Sphingosine kinase 1 (SphK1), a key enzyme for S1P production, is concentrated in presynaptic terminals. However, the role of S1P/SphK1 signaling in exocytosis remains elusive. By detecting catecholamine release from single vesicles in chromaffin cells, we show that a dominant negative SphK1 (SphK1
DN ) reduces the number of amperometric spikes and increases the duration of foot, which reflects release through a fusion pore, implying critical roles for S1P in regulating the rate of exocytosis and fusion pore expansion. Similar phenotypes were observed in chromaffin cells obtained from SphK1 knockout mice compared to those from wild-type mice. In addition, extracellular S1P treatment increased the number of amperometric spikes, and this increase, in turn, was inhibited by a selective S1P3 receptor blocker, suggesting extracellular S1P may regulate the rate of exocytosis via activation of S1P3. Furthermore, intracellular S1P application induced a decrease in foot duration of amperometric spikes in control cells, indicating intracellular S1P may regulate fusion pore expansion during exocytosis. Taken together, our study represents the first demonstration that S1P regulates exocytosis through distinct mechanisms: extracellular S1P may modulate the rate of exocytosis via activation of S1P receptors while intracellular S1P may directly control fusion pore expansion during exocytosis. OPEN SCIENCE BADGES: This article has received a badge for *Open Materials* because it provided all relevant information to reproduce the study in the manuscript. The complete Open Science Disclosure form for this article can be found at the end of the article. More information about the Open Practices badges can be found at https://cos.io/our-services/open-science-badges/., (© 2019 International Society for Neurochemistry.)- Published
- 2019
- Full Text
- View/download PDF
35. Abundance of Synaptic Vesicle-Related Proteins in Alpha-Synuclein-Containing Protein Inclusions Suggests a Targeted Formation Mechanism.
- Author
-
McCormack A, Keating DJ, Chegeni N, Colella A, Wang JJ, and Chataway T
- Subjects
- Aged, Aged, 80 and over, Female, Humans, Lewy Bodies metabolism, Male, Middle Aged, Oligodendroglia metabolism, Protein Aggregation, Pathological metabolism, Inclusion Bodies metabolism, Lewy Body Disease metabolism, Multiple System Atrophy metabolism, Synaptic Vesicles metabolism, alpha-Synuclein metabolism
- Abstract
Proteinaceous α-synuclein-containing inclusions are found in affected brain regions in patients with Parkinson's disease (PD), Dementia with Lewy bodies (DLB) and multiple system atrophy (MSA). These appear in neurons as Lewy bodies in both PD and DLB and as glial cytoplasmic inclusions (GCIs) in oligodendrocytes in MSA. The role they play in the pathology of the diseases is unknown, and relatively little is still known about their composition. By purifying the inclusions from the surrounding tissue and comprehensively analysing their protein composition, vital clues to the formation mechanism and role in the disease process may be found. In this study, Lewy bodies were purified from postmortem brain tissue from DLB cases (n = 2) and GCIs were purified from MSA cases (n = 5) using a recently improved purification method, and the purified inclusions were analysed by mass spectrometry. Twenty-one percent of the proteins found consistently in the GCIs and LBs were synaptic-vesicle related. Identified proteins included those associated with exosomes (CD9), clathrin-mediated endocytosis (clathrin, AP-2 complex, dynamin), retrograde transport (dynein, dynactin, spectrin) and synaptic vesicle fusion (synaptosomal-associated protein 25, vesicle-associated membrane protein 2, syntaxin-1). This suggests that the misfolded or excess α-synuclein may be targeted to inclusions via vesicle-mediated transport, which also explains the presence of the neuronal protein α-synuclein within GCIs.
- Published
- 2019
- Full Text
- View/download PDF
36. The Influence of the Gut Microbiome on Host Metabolism Through the Regulation of Gut Hormone Release.
- Author
-
Martin AM, Sun EW, Rogers GB, and Keating DJ
- Abstract
The microbial community of the gut conveys significant benefits to host physiology. A clear relationship has now been established between gut bacteria and host metabolism in which microbial-mediated gut hormone release plays an important role. Within the gut lumen, bacteria produce a number of metabolites and contain structural components that act as signaling molecules to a number of cell types within the mucosa. Enteroendocrine cells within the mucosal lining of the gut synthesize and secrete a number of hormones including CCK, PYY, GLP-1, GIP, and 5-HT, which have regulatory roles in key metabolic processes such as insulin sensitivity, glucose tolerance, fat storage, and appetite. Release of these hormones can be influenced by the presence of bacteria and their metabolites within the gut and as such, microbial-mediated gut hormone release is an important component of microbial regulation of host metabolism. Dietary or pharmacological interventions which alter the gut microbiome therefore pose as potential therapeutics for the treatment of human metabolic disorders. This review aims to describe the complex interaction between intestinal microbiota and their metabolites and gut enteroendocrine cells, and highlight how the gut microbiome can influence host metabolism through the regulation of gut hormone release.
- Published
- 2019
- Full Text
- View/download PDF
37. What is the role of endogenous gut serotonin in the control of gastrointestinal motility?
- Author
-
Keating DJ and Spencer NJ
- Subjects
- Animals, Gastrointestinal Tract physiology, Humans, Gastrointestinal Motility physiology, Serotonin physiology
- Abstract
In recent years, there have been dramatic changes in our understanding of the role of endogenous 5-Hydroxytryptamine (5-HT or serotonin) in the control of gastrointestinal (GI) motility. Whilst it is well accepted that there are numerous types of 5-HT receptors expressed on enteric neurons and that exogenous 5-HT potently stimulates GI-motility, understanding the role of endogenous 5-HT in GI-motility has been substantially more difficult to resolve. Recent studies found 5-HT
3 and 5-HT4 antagonists have the same effects on peristalsis in colon preparations depleted of endogenous 5-HT. Then, recent work revealed that in mice with genetic mutations to prevent the synthesis of endogenous 5-HT from enterochromaffin EC) cells did not block major neurogenic motor patterns in the gut wall and did not reduce GI-transit in conscious animals, raising doubts about early hypotheses that endogenous 5-HT was critical for neurogenic GI-motility patterns. Indeed, functional evidence now suggests that 5-HT3 and 5-HT4 receptors on enteric nerves display constitutive activity. In summary, recent findings demonstrate that endogenous 5-HT released from the mucosa or enteric neurons is not required for the generation of major neurogenic motor patterns, at least in the large intestine, but that it likely acts as a modulator of contractile frequency. This review will discuss how and why our understanding of endogenous 5-HT has dramatically changed in the past few years., (Copyright © 2018 Elsevier Ltd. All rights reserved.)- Published
- 2019
- Full Text
- View/download PDF
38. Sugar Responses of Human Enterochromaffin Cells Depend on Gut Region, Sex, and Body Mass.
- Author
-
Lumsden AL, Martin AM, Sun EW, Schober G, Isaacs NJ, Pezos N, Wattchow DA, de Fontgalland D, Rabbitt P, Hollington P, Sposato L, Due SL, Rayner CK, Nguyen NQ, Liou AP, Jackson VM, Young RL, and Keating DJ
- Subjects
- Cells, Cultured, Dose-Response Relationship, Drug, Female, Humans, Male, Sex Factors, Body Weight, Carbohydrates pharmacology, Enterochromaffin Cells drug effects, Gastrointestinal Tract cytology
- Abstract
Gut-derived serotonin (5-HT) is released from enterochromaffin (EC) cells in response to nutrient cues, and acts to slow gastric emptying and modulate gastric motility. Rodent studies also evidence a role for gut-derived 5-HT in the control of hepatic glucose production, lipolysis and thermogenesis, and in mediating diet-induced obesity. EC cell number and 5-HT content is increased in the small intestine of obese rodents and human, however, it is unknown whether EC cells respond directly to glucose in humans, and whether their capacity to release 5-HT is perturbed in obesity. We therefore investigated 5-HT release from human duodenal and colonic EC cells in response to glucose, sucrose, fructose and α-glucoside (αMG) in relation to body mass index (BMI). EC cells released 5-HT only in response to 100 and 300 mM glucose (duodenum) and 300 mM glucose (colon), independently of osmolarity. Duodenal, but not colonic, EC cells also released 5-HT in response to sucrose and αMG, but did not respond to fructose. 5-HT content was similar in all EC cells in males, and colonic EC cells in females, but 3 to 4-fold higher in duodenal EC cells from overweight females ( p < 0.05 compared to lean, obese). Glucose-evoked 5-HT release was 3-fold higher in the duodenum of overweight females ( p < 0.05, compared to obese), but absent here in overweight males. Our data demonstrate that primary human EC cells respond directly to dietary glucose cues, with regional differences in selectivity for other sugars. Augmented glucose-evoked 5-HT release from duodenal EC is a feature of overweight females, and may be an early determinant of obesity.
- Published
- 2019
- Full Text
- View/download PDF
39. The Regulation of Peripheral Metabolism by Gut-Derived Hormones.
- Author
-
Sun EWL, Martin AM, Young RL, and Keating DJ
- Abstract
Enteroendocrine cells lining the gut epithelium constitute the largest endocrine organ in the body and secrete over 20 different hormones in response to cues from ingested foods and changes in nutritional status. Not only do these hormones convey signals from the gut to the brain via the gut-brain axis, they also act directly on metabolically important peripheral targets in a highly concerted fashion to maintain energy balance and glucose homeostasis. Gut-derived hormones released during fasting tend to be orexigenic and have hyperglycaemic potential. Conversely, gut hormones secreted postprandially generally promote satiety and facilitate glucose clearance. Although some of the metabolic benefits conferred by bariatric surgeries have been ascribed to changes in the secretory profiles of various gut hormones, the therapeutic potential of the enteroendocrine system as a viable target against metabolic diseases remain largely underexploited, except for incretin-mimetics. This review provides a brief overview of the physiological importance and highlights the therapeutic potential of the following gut hormones: serotonin, glucose-dependent insulinotropic peptide, glucagon-like peptide 1, oxyntomodulin, peptide YY, insulin-like peptide 5, and ghrelin.
- Published
- 2019
- Full Text
- View/download PDF
40. Metformin-induced glucagon-like peptide-1 secretion contributes to the actions of metformin in type 2 diabetes.
- Author
-
Bahne E, Sun EWL, Young RL, Hansen M, Sonne DP, Hansen JS, Rohde U, Liou AP, Jackson ML, de Fontgalland D, Rabbitt P, Hollington P, Sposato L, Due S, Wattchow DA, Rehfeld JF, Holst JJ, Keating DJ, Vilsbøll T, and Knop FK
- Subjects
- Adult, Aged, Aged, 80 and over, Australia, Blood Glucose drug effects, Double-Blind Method, Female, Humans, Male, Middle Aged, Postprandial Period, Diabetes Mellitus, Type 2 metabolism, Glucagon-Like Peptide 1 metabolism, Metformin pharmacology
- Abstract
Background: Metformin reduces plasma glucose and has been shown to increase glucagon-like peptide 1 (GLP-1) secretion. Whether this is a direct action of metformin on GLP-1 release, and whether some of the glucose-lowering effect of metformin occurs due to GLP-1 release, is unknown. The current study investigated metformin-induced GLP-1 secretion and its contribution to the overall glucose-lowering effect of metformin and underlying mechanisms in patients with type 2 diabetes., Methods: Twelve patients with type 2 diabetes were included in this placebo-controlled, double-blinded study. On 4 separate days, the patients received metformin (1,500 mg) or placebo suspended in a liquid meal, with subsequent i.v. infusion of the GLP-1 receptor antagonist exendin9-39 (Ex9-39) or saline. During 240 minutes, blood was sampled. The direct effect of metformin on GLP-1 secretion was tested ex vivo in human ileal and colonic tissue with and without dorsomorphin-induced inhibiting of the AMPK activity., Results: Metformin increased postprandial GLP-1 secretion compared with placebo (P = 0.014), and the postprandial glucose excursions were significantly smaller after metformin + saline compared with metformin + Ex9-39 (P = 0.004). Ex vivo metformin acutely increased GLP-1 secretion (colonic tissue, P < 0.01; ileal tissue, P < 0.05), but the effect was abolished by inhibition of AMPK activity., Conclusions: Metformin has a direct and AMPK-dependent effect on GLP-1-secreting L cells and increases postprandial GLP-1 secretion, which seems to contribute to metformin's glucose-lowering effect and mode of action., Trial Registration: NCT02050074 (https://clinicaltrials.gov/ct2/show/NCT02050074)., Funding: This study received grants from the A.P. Møller Foundation, the Novo Nordisk Foundation, the Danish Medical Association research grant, the Australian Research Council, the National Health and Medical Research Council, and Pfizer Inc.
- Published
- 2018
- Full Text
- View/download PDF
41. Gut Mechanisms Linking Intestinal Sweet Sensing to Glycemic Control.
- Author
-
Kreuch D, Keating DJ, Wu T, Horowitz M, Rayner CK, and Young RL
- Abstract
Sensing nutrients within the gastrointestinal tract engages the enteroendocrine cell system to signal within the mucosa, to intrinsic and extrinsic nerve pathways, and the circulation. This signaling provides powerful feedback from the intestine to slow the rate of gastric emptying, limit postprandial glycemic excursions, and induce satiation. This review focuses on the intestinal sensing of sweet stimuli (including low-calorie sweeteners), which engage similar G-protein-coupled receptors (GPCRs) to the sweet taste receptors (STRs) of the tongue. It explores the enteroendocrine cell signals deployed upon STR activation that act within and outside the gastrointestinal tract, with a focus on the role of this distinctive pathway in regulating glucose transport function via absorptive enterocytes, and the associated impact on postprandial glycemic responses in animals and humans. The emerging role of diet, including low-calorie sweeteners, in modulating the composition of the gut microbiome and how this may impact glycemic responses of the host, is also discussed, as is recent evidence of a causal role of diet-induced dysbiosis in influencing the gut-brain axis to alter gastric emptying and insulin release. Full knowledge of intestinal STR signaling in humans, and its capacity to engage host and/or microbiome mechanisms that modify glycemic control, holds the potential for improved prevention and management of type 2 diabetes.
- Published
- 2018
- Full Text
- View/download PDF
42. Regulator of Calcineurin 1 helps coordinate whole-body metabolism and thermogenesis.
- Author
-
Rotter D, Peiris H, Grinsfelder DB, Martin AM, Burchfield J, Parra V, Hull C, Morales CR, Jessup CF, Matusica D, Parks BW, Lusis AJ, Nguyen NUN, Oh M, Iyoke I, Jakkampudi T, McMillan DR, Sadek HA, Watt MJ, Gupta RK, Pritchard MA, Keating DJ, and Rothermel BA
- Subjects
- 3T3-L1 Cells, Adipocytes cytology, Adipocytes drug effects, Adipocytes metabolism, Adipose Tissue metabolism, Adipose Tissue, Beige drug effects, Adipose Tissue, Beige metabolism, Adipose Tissue, White drug effects, Adipose Tissue, White metabolism, Adrenergic Agents pharmacology, Animals, Calcineurin metabolism, Calcium-Binding Proteins, Cell Differentiation drug effects, Cold Temperature, Female, Insulin Resistance, Intracellular Signaling Peptides and Proteins deficiency, Lipid Metabolism drug effects, Liver metabolism, Male, Metabolic Syndrome metabolism, Mice, Mice, Knockout, Muscle Proteins deficiency, Muscle Proteins genetics, Muscle, Skeletal metabolism, Muscle, Striated metabolism, Obesity metabolism, Obesity pathology, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha metabolism, Promoter Regions, Genetic genetics, Proteolipids genetics, Proteolipids metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Uncoupling Protein 1 metabolism, Intracellular Signaling Peptides and Proteins metabolism, Metabolism drug effects, Muscle Proteins metabolism, Thermogenesis drug effects
- Abstract
Increasing non-shivering thermogenesis (NST), which expends calories as heat rather than storing them as fat, is championed as an effective way to combat obesity and metabolic disease. Innate mechanisms constraining the capacity for NST present a fundamental limitation to this approach, yet are not well understood. Here, we provide evidence that Regulator of Calcineurin 1 ( RCAN1 ), a feedback inhibitor of the calcium-activated protein phosphatase calcineurin (CN), acts to suppress two distinctly different mechanisms of non-shivering thermogenesis (NST): one involving the activation of UCP1 expression in white adipose tissue, the other mediated by sarcolipin (SLN) in skeletal muscle. UCP1 generates heat at the expense of reducing ATP production, whereas SLN increases ATP consumption to generate heat. Gene expression profiles demonstrate a high correlation between Rcan1 expression and metabolic syndrome. On an evolutionary timescale, in the context of limited food resources, systemic suppression of prolonged NST by RCAN1 might have been beneficial; however, in the face of caloric abundance, RCAN1-mediated suppression of these adaptive avenues of energy expenditure may now contribute to the growing epidemic of obesity., (© 2018 The Authors.)
- Published
- 2018
- Full Text
- View/download PDF
43. Augmented capacity for peripheral serotonin release in human obesity.
- Author
-
Young RL, Lumsden AL, Martin AM, Schober G, Pezos N, Thazhath SS, Isaacs NJ, Cvijanovic N, Sun EWL, Wu T, Rayner CK, Nguyen NQ, Fontgalland D, Rabbitt P, Hollington P, Sposato L, Due SL, Wattchow DA, Liou AP, Jackson VM, and Keating DJ
- Subjects
- Adult, Blood Glucose metabolism, Cells, Cultured, Colon metabolism, Endoscopy, Gastrointestinal, Female, Humans, Male, Middle Aged, Obesity metabolism, Peripheral Nervous System metabolism, Real-Time Polymerase Chain Reaction, Signal Transduction, Colon cytology, Enterochromaffin Cells metabolism, Obesity physiopathology, Peripheral Nervous System physiology, Serotonin metabolism
- Abstract
Background/objectives: Evidence from animal studies highlights an important role for serotonin (5-HT), derived from gut enterochromaffin (EC) cells, in regulating hepatic glucose production, lipolysis and thermogenesis, and promoting obesity and dysglycemia. Evidence in humans is limited, although elevated plasma 5-HT concentrations are linked to obesity., Subjects/methods: We assessed (i) plasma 5-HT concentrations before and during intraduodenal glucose infusion (4 kcal/min for 30 min) in non-diabetic obese (BMI 44 ± 4 kg/m
2 , N = 14) and control (BMI 24 ± 1 kg/m2 , N = 10) subjects, (ii) functional activation of duodenal EC cells (immunodetection of phospho-extracellular related-kinase, pERK) in response to glucose, and in separate subjects, (iii) expression of tryptophan hydroxylase-1 (TPH1) in duodenum and colon (N = 39), and (iv) 5-HT content in primary EC cells from these regions (N = 85)., Results: Plasma 5-HT was twofold higher in obese than control responders prior to (P = 0.025), and during (iAUC, P = 0.009), intraduodenal glucose infusion, and related positively to BMI (R2 = 0.334, P = 0.003) and HbA1c (R2 = 0.508, P = 0.009). The density of EC cells in the duodenum was twofold higher at baseline in obese subjects than controls (P = 0.023), with twofold more EC cells activated by glucose infusion in the obese (EC cells co-expressing 5-HT and pERK, P = 0.001), while the 5-HT content of EC cells in duodenum and colon was similar; TPH1 expression was 1.4-fold higher in the duodenum of obese subjects (P = 0.044), and related positively to BMI (R2 = 0.310, P = 0.031)., Conclusions: Human obesity is characterized by an increased capacity to produce and release 5-HT from the proximal small intestine, which is strongly linked to higher body mass, and glycemic control. Gut-derived 5-HT is likely to be an important driver of pathogenesis in human obesity and dysglycemia.- Published
- 2018
- Full Text
- View/download PDF
44. Identification of a Rhythmic Firing Pattern in the Enteric Nervous System That Generates Rhythmic Electrical Activity in Smooth Muscle.
- Author
-
Spencer NJ, Hibberd TJ, Travis L, Wiklendt L, Costa M, Hu H, Brookes SJ, Wattchow DA, Dinning PG, Keating DJ, and Sorensen J
- Subjects
- Animals, Female, Intestines innervation, Intestines physiology, Male, Mice, Mice, Inbred C57BL, Neuroimaging methods, Enteric Nervous System physiology, Muscle, Smooth physiology, Myoelectric Complex, Migrating physiology
- Abstract
The enteric nervous system (ENS) contains millions of neurons essential for organization of motor behavior of the intestine. It is well established that the large intestine requires ENS activity to drive propulsive motor behaviors. However, the firing pattern of the ENS underlying propagating neurogenic contractions of the large intestine remains unknown. To identify this, we used high-resolution neuronal imaging with electrophysiology from neighboring smooth muscle. Myoelectric activity underlying propagating neurogenic contractions along murine large intestine [also referred to as colonic migrating motor complexes, (CMMCs)] consisted of prolonged bursts of rhythmic depolarizations at a frequency of ∼2 Hz. Temporal coordination of this activity in the smooth muscle over large spatial fields (∼7 mm, longitudinally) was dependent on the ENS. During quiescent periods between neurogenic contractions, recordings from large populations of enteric neurons, in mice of either sex, revealed ongoing activity. The onset of neurogenic contractions was characterized by the emergence of temporally synchronized activity across large populations of excitatory and inhibitory neurons. This neuronal firing pattern was rhythmic and temporally synchronized across large numbers of ganglia at ∼2 Hz. ENS activation preceded smooth muscle depolarization, indicating rhythmic depolarizations in smooth muscle were controlled by firing of enteric neurons. The cyclical emergence of temporally coordinated firing of large populations of enteric neurons represents a unique neural motor pattern outside the CNS. This is the first direct observation of rhythmic firing in the ENS underlying rhythmic electrical depolarizations in smooth muscle. The pattern of neuronal activity we identified underlies the generation of CMMCs. SIGNIFICANCE STATEMENT How the enteric nervous system (ENS) generates neurogenic contractions of smooth muscle in the gastrointestinal (GI) tract has been a long-standing mystery in vertebrates. It is well known that myogenic pacemaker cells exist in the GI tract [called interstitial cells of Cajal (ICCs)] that generate rhythmic myogenic contractions. However, the mechanisms underlying the generation of rhythmic neurogenic contractions of smooth muscle in the GI tract remains unknown. We developed a high-resolution neuronal imaging method with electrophysiology to address this issue. This technique revealed a novel pattern of rhythmic coordinated neuronal firing in the ENS that has never been identified. Rhythmic neuronal firing in the ENS was found to generate rhythmic neurogenic depolarizations in smooth muscle that underlie contraction of the GI tract., (Copyright © 2018 the authors 0270-6474/18/385508-16$15.00/0.)
- Published
- 2018
- Full Text
- View/download PDF
45. ICAM-1-related long non-coding RNA: promoter analysis and expression in human retinal endothelial cells.
- Author
-
Lumsden AL, Ma Y, Ashander LM, Stempel AJ, Keating DJ, Smith JR, and Appukuttan B
- Subjects
- Alleles, Binding Sites, Humans, Intercellular Adhesion Molecule-1 metabolism, Phenotype, Polymorphism, Single Nucleotide genetics, Protein Binding, RNA, Long Noncoding metabolism, Transcription Factors metabolism, Endothelial Cells metabolism, Gene Expression Regulation, Intercellular Adhesion Molecule-1 genetics, Promoter Regions, Genetic, RNA, Long Noncoding genetics, Retina cytology
- Abstract
Objective: Regulation of intercellular adhesion molecule (ICAM)-1 in retinal endothelial cells is a promising druggable target for retinal vascular diseases. The ICAM-1-related (ICR) long non-coding RNA stabilizes ICAM-1 transcript, increasing protein expression. However, studies of ICR involvement in disease have been limited as the promoter is uncharacterized. To address this issue, we undertook a comprehensive in silico analysis of the human ICR gene promoter region., Results: We used genomic evolutionary rate profiling to identify a 115 base pair (bp) sequence within 500 bp upstream of the transcription start site of the annotated human ICR gene that was conserved across 25 eutherian genomes. A second constrained sequence upstream of the orthologous mouse gene (68 bp; conserved across 27 Eutherian genomes including human) was also discovered. Searching these elements identified 33 matrices predictive of binding sites for transcription factors known to be responsive to a broad range of pathological stimuli, including hypoxia, and metabolic and inflammatory proteins. Five phenotype-associated single nucleotide polymorphisms (SNPs) in the immediate vicinity of these elements included four SNPs (i.e. rs2569693, rs281439, rs281440 and rs11575074) predicted to impact binding motifs of transcription factors, and thus the expression of ICR and ICAM-1 genes, with potential to influence disease susceptibility. We verified that human retinal endothelial cells expressed ICR, and observed induction of expression by tumor necrosis factor-α.
- Published
- 2018
- Full Text
- View/download PDF
46. The neuronal and endocrine roles of RCAN1 in health and disease.
- Author
-
Peiris H and Keating DJ
- Subjects
- Animals, Humans, Disease, Endocrine System metabolism, Health, Intracellular Signaling Peptides and Proteins metabolism, Neurons metabolism
- Abstract
The regulator of calcineurin 1 (RCAN1) was first discovered as a gene located on human chromosome 21, expressed in neurons and overexpressed in the brains of Down syndrome individuals. Increased expression of RCAN1 has been linked with not only Down syndrome-associated pathology but also an associated neurological disorder, Alzheimer's Disease, in which neuronal RCAN1 expression is also increased. RCAN1 has additionally been demonstrated to affect other cell types including endocrine cells, with links to the pathogenesis of β-cell dysfunction in type 2 diabetes. The primary functions of RCAN1 relate to the inhibition of the phosphatase calcineurin, and to the regulation of mitochondrial function. Various forms of cellular stress such as reactive oxygen species and hyperglycaemia cause transient increases in RCAN1 expression. The short term (hours to days) induction of RCAN1 expression is generally thought to have a protective effect by regulating the expression of pro-survival genes in multiple cell types, many of which are mediated via the calcineurin/NFAT transcriptional pathway. However, strong evidence also supports the notion that chronic (weeks-years) overexpression of RCAN1 has a detrimental effect on cells and that this may drive pathophysiological changes in neurons and endocrine cells linked to Down syndrome, Alzheimer's Disease and type 2 diabetes. Here we review the evidence related to these roles of RCAN1 in neurons and endocrine cells and their relationship to these human health disorders., (© 2017 John Wiley & Sons Australia, Ltd.)
- Published
- 2018
- Full Text
- View/download PDF
47. p75 neurotrophin receptor interacts with and promotes BACE1 localization in endosomes aggravating amyloidogenesis.
- Author
-
Saadipour K, Mañucat-Tan NB, Lim Y, Keating DJ, Smith KS, Zhong JH, Liao H, Bobrovskaya L, Wang YJ, Chao MV, and Zhou XF
- Subjects
- Amyloid beta-Protein Precursor metabolism, Animals, Disease Models, Animal, Mice, Knockout, Primary Cell Culture, Receptors, Nerve Growth Factor genetics, Signal Transduction, Alzheimer Disease metabolism, Amyloid Precursor Protein Secretases metabolism, Amyloid beta-Peptides metabolism, Aspartic Acid Endopeptidases metabolism, Cerebral Cortex metabolism, Endosomes metabolism, Neurons metabolism, Receptors, Nerve Growth Factor metabolism
- Abstract
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by a progressive deposition of amyloid beta (Aβ) and dysregulation of neurotrophic signaling, causing synaptic dysfunction, loss of memory, and cell death. The expression of p75 neurotrophin receptor is elevated in the brain of AD patients, suggesting its involvement in this disease. However, the exact mechanism of its action is not yet clear. Here, we show that p75 interacts with beta-site amyloid precursor protein cleaving enzyme-1 (BACE1), and this interaction is enhanced in the presence of Aβ. Our results suggest that the colocalization of BACE1 and amyloid precursor protein (APP) is increased in the presence of both Aβ and p75 in cortical neurons. In addition, the localization of APP and BACE1 in early endosomes is increased in the presence of Aβ and p75. An increased phosphorylation of APP-Thr668 and BACE1-Ser498 by c-Jun N-terminal kinase (JNK) in the presence of Aβ and p75 could be responsible for this localization. In conclusion, our study proposes a potential involvement in amyloidogenesis for p75, which may represent a future therapeutic target for AD. Cover Image for this Issue: doi. 10.1111/jnc.14163., (© 2017 International Society for Neurochemistry.)
- Published
- 2018
- Full Text
- View/download PDF
48. Synaptic activation of putative sensory neurons by hexamethonium-sensitive nerve pathways in mouse colon.
- Author
-
Hibberd TJ, Travis L, Wiklendt L, Costa M, Brookes SJH, Hu H, Keating DJ, and Spencer NJ
- Subjects
- Animals, Calcitonin Gene-Related Peptide metabolism, Calcium Signaling drug effects, Electric Stimulation, Evoked Potentials drug effects, Female, In Vitro Techniques, Kinetics, Male, Mice, Inbred C57BL, Myenteric Plexus metabolism, Nitric Oxide Synthase Type I metabolism, Reaction Time, Sensory Receptor Cells metabolism, Colon innervation, Hexamethonium pharmacology, Myenteric Plexus drug effects, Nicotinic Antagonists pharmacology, Sensory Receptor Cells drug effects, Synaptic Transmission drug effects
- Abstract
The gastrointestinal tract contains its own independent population of sensory neurons within the gut wall. These sensory neurons have been referred to as intrinsic primary afferent neurons (IPANs) and can be identified by immunoreactivity to calcitonin gene-related peptide (CGRP) in mice. A common feature of IPANs is a paucity of fast synaptic inputs observed during sharp microelectrode recordings. Whether this is observed using different recording techniques is of particular interest for understanding the physiology of these neurons and neural circuit modeling. Here, we imaged spontaneous and evoked activation of myenteric neurons in isolated whole preparations of mouse colon and correlated recordings with CGRP and nitric oxide synthase (NOS) immunoreactivity, post hoc. Calcium indicator fluo 4 was used for this purpose. Calcium responses were recorded in nerve cell bodies located 5-10 mm oral to transmural electrical nerve stimuli. A total of 618 recorded neurons were classified for CGRP or NOS immunoreactivity. Aboral electrical stimulation evoked short-latency calcium transients in the majority of myenteric neurons, including ~90% of CGRP-immunoreactive Dogiel type II neurons. Activation of Dogiel type II neurons had a time course consistent with fast synaptic transmission and was always abolished by hexamethonium (300 μM) and by low-calcium Krebs solution. The nicotinic receptor agonist 1,1-dimethyl-4-phenylpiperazinium iodide (during synaptic blockade) directly activated Dogiel type II neurons. The present study suggests that murine colonic Dogiel type II neurons receive prominent fast excitatory synaptic inputs from hexamethonium-sensitive neural pathways. NEW & NOTEWORTHY Myenteric neurons in isolated mouse colon were recorded using calcium imaging and then neurochemically defined. Short-latency calcium transients were detected in >90% of calcitonin gene-related peptide-immunoreactive neurons to electrical stimulation of hexamethonium-sensitive pathways. Putative sensory Dogiel type II calcitonin gene-related peptide-immunoreactive myenteric neurons may receive widespread fast synaptic inputs in mouse colon.
- Published
- 2018
- Full Text
- View/download PDF
49. Current Therapies That Modify Glucagon Secretion: What Is the Therapeutic Effect of Such Modifications?
- Author
-
Grøndahl MF, Keating DJ, Vilsbøll T, and Knop FK
- Subjects
- Humans, Hypoglycemic Agents therapeutic use, Molecular Targeted Therapy, Receptors, Glucagon metabolism, Diabetes Mellitus, Type 2 drug therapy, Glucagon metabolism
- Abstract
Purpose of Review: Hyperglucagonemia contributes significantly to hyperglycemia in type 2 diabetes and suppressed glucagon levels may increase the risk of hypoglycemia. Here, we give a brief overview of glucagon physiology and the role of glucagon in the pathophysiology of type 2 diabetes and provide insights into how antidiabetic drugs influence glucagon secretion as well as a perspective on the future of glucagon-targeting drugs., Recent Findings: Several older as well as recent investigations have evaluated the effect of antidiabetic agents on glucagon secretion to understand how glucagon may be involved in the drugs' efficacy and safety profiles. Based on these findings, modulation of glucagon secretion seems to play a hitherto underestimated role in the efficacy and safety of several glucose-lowering drugs. Numerous drugs currently available to diabetologists are capable of altering glucagon secretion: metformin, sulfonylurea compounds, insulin, glucagon-like peptide-1 receptor agonists, dipeptidyl peptidase-4 inhibitors, sodium-glucose cotransporter 2 inhibitors and amylin mimetics. Their diverse effects on glucagon secretion are of importance for their individual efficacy and safety profiles. Understanding how these drugs interact with glucagon secretion may help to optimize treatment.
- Published
- 2017
- Full Text
- View/download PDF
50. Inhibition of Miro1 disturbs mitophagy and pancreatic β-cell function interfering insulin release via IRS-Akt-Foxo1 in diabetes.
- Author
-
Chen L, Liu C, Gao J, Xie Z, Chan LWC, Keating DJ, Yang Y, Sun J, Zhou F, Wei Y, Men X, and Yang S
- Abstract
Mitochondrial function is essential to meet metabolic demand of pancreatic beta cells respond to high nutrient stress. Mitophagy is an essential component to normal pancreatic β-cell function and has been associated with β-cell failure in Type 2 diabetes (T2D). Our previous studies have indicated that mitochondrial Rho (Miro) GTPase-mediated mitochondrial dysfunction under high nutrient stress leads to NOD-like receptor 3 (NLRP3)-dependent proinflammatory responses and subsequent insulin resistance. However, the in vivo mechanism by which Miro1 underlies mitophagy has not been identified. Here we show firstly that the expression of Miro is reduced in human T2D and mouse db/db islets and in INS-1 cell line exposed to high glucose and palmitate. β-cell specific ablation of Miro1 (Miro1f/f: Rip-cre mice, or (IKO) under high nutrient stress promotes the development of hyperglycemia. β-cells from IKO mice display an inhibition of mitophagy under oxidative stress and induces mitochondrial dysfunction. Dysfunctional mitophagy in IKO mice is represented by damaged islet beta cell mitochondrial and secretory capacity, unbalanced downstream MKK-JNK signalling without affecting the levels of MEK, ERK or p38 activation and subsequently, impaired insulin secretion signaling via inhibition IRS-AKT-Foxo1 pathway, leading to worsening glucose tolerance in these mice. Thus, these data suggest that Miro1 may be responsible for mitophagy deficiency and β-cell dysfunction in T2D and that strategies target Miro1 in vivo may provide a therapeutic target to enhance β-cell mitochondrial quality and insulin secretion to ameliorate complications associated with T2D., Competing Interests: CONFLICTS OF INTEREST The authors declare no competing financial interests.
- Published
- 2017
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.