96 results on '"Leibiger I"'
Search Results
2. Single residue (K332A) substitution in Kir6.2 abolishes the stimulatory effect of long-chain acyl-CoA esters: indications for a long-chain acyl-CoA ester binding motif
- Author
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Bränström, R., Leibiger, I. B., Leibiger, B., Klement, G., Nilsson, J., Århem, P., Aspinwall, C. A., Corkey, B. E., Larsson, O., and Berggren, P.-O.
- Published
- 2007
- Full Text
- View/download PDF
3. Proinsulin C-peptide and insulin: Limited pattern similarities of interest in inter-peptide interactions but no C-peptide effect on insulin and IGF-1 receptor signaling
- Author
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Henriksson, M., Johansson, J., Moede, T., Leibiger, I., Shafqat, J., Berggren, P. -O., and Jörnvall, H.
- Published
- 2006
- Full Text
- View/download PDF
4. Non-invasive in vivo imaging of pancreatic β-cell function and survival – a perspective
- Author
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Leibiger, I. B., Caicedo, A., and Berggren, P.-O.
- Published
- 2012
- Full Text
- View/download PDF
5. A Novel Assay for Detection of Amyloid Induced Apoptosis
- Author
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Schultz, S, primary, Berggren, P, additional, Westermark, G, additional, Leibiger, I, additional, Paulsson, J, additional, and Köhler, M, additional
- Published
- 2007
- Full Text
- View/download PDF
6. Pals-associated tight junction protein functionally links dopamine and angiotensin II to the regulation of sodium transport in renal epithelial cells
- Author
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Chen, Z, Leibiger, I, Katz, A I, and Bertorello, A M
- Published
- 2009
- Full Text
- View/download PDF
7. Dual function of the intron of the rat insulin I gene in regulation of gene expression
- Author
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Damert, A., Leibiger, B., and Leibiger, I. B.
- Published
- 1996
- Full Text
- View/download PDF
8. Functional analysis of DNA-elements involved in transcriptional control of the human glucose transporter 2 (GLUT 2) gene in the insulin-producing cell line βTC-3
- Author
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Leibiger, B. and Leibiger, I. B.
- Published
- 1995
- Full Text
- View/download PDF
9. Non-invasive in vivo imaging of pancreatic β-cell function and survival - a perspective
- Author
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Leibiger, I. B., Caicedo, A., and Berggren, P.-O.
- Published
- 2011
- Full Text
- View/download PDF
10. Molecular cloning of a novel putative murine mitochondrial transporter
- Author
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Li, F.Y., Leibiger, B., Leibiger, I., and Larsson, C.
- Subjects
Cloning -- Research ,Mitochondria -- Genetic aspects ,Gene expression -- Research ,Biological sciences - Published
- 2001
11. Loss of parafibromin expression in a subset of parathyroid adenomas
- Author
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Juhlin, C, primary, Larsson, C, additional, Yakoleva, T, additional, Leibiger, I, additional, Leibiger, B, additional, Alimov, A, additional, Weber, G, additional, Höög, A, additional, and Villablanca, A, additional
- Published
- 2006
- Full Text
- View/download PDF
12. Inositol hexakisphosphate promotes dynamin I- mediated endocytosis
- Author
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Hoy, M., primary, Efanov, A. M., additional, Bertorello, A. M., additional, Zaitsev, S. V., additional, Olsen, H. L., additional, Bokvist, K., additional, Leibiger, B., additional, Leibiger, I. B., additional, Zwiller, J., additional, Berggren, P.-O., additional, and Gromada, J., additional
- Published
- 2002
- Full Text
- View/download PDF
13. Short-term regulation of insulin gene transcription
- Author
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Leibiger, B., primary, Moede, T., additional, Uhles, S., additional, Berggren, P.-O., additional, and Leibiger, I. B., additional
- Published
- 2002
- Full Text
- View/download PDF
14. Online monitoring of stimulus-induced gene expression in pancreatic beta-cells.
- Author
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Moede, T, primary, Leibiger, B, additional, Berggren, P O, additional, and Leibiger, I B, additional
- Published
- 2001
- Full Text
- View/download PDF
15. Overexpression of cytosolically-directed Multiple Inositol Polyphosphate Phosphatase in insulin secreting cells
- Author
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Barker, C. J., primary, Yu, J., additional, Leibiger, B., additional, Leibiger, I., additional, Caffrey, J. J., additional, Shears, S. B., additional, and Berggren, P. O., additional
- Published
- 2000
- Full Text
- View/download PDF
16. Functional analysis of DNA-elements involved in transcriptional control of the human glucose transporter 2 (GLUT 2) gene in the insulin-producing cell line ?TC-3
- Author
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Leibiger, B., primary and Leibiger, I. B., additional
- Published
- 1995
- Full Text
- View/download PDF
17. The neuropeptide galanin occurs in two conformations
- Author
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Lindskog, S., primary, Ahrén, B., additional, Leibiger, I., additional, and Zühlke, H., additional
- Published
- 1992
- Full Text
- View/download PDF
18. Long chain coenzyme A esters activate the pore-forming subunit (Kir6. 2) of the ATP-regulated potassium channel.
- Author
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Bränström, R, Leibiger, I B, Leibiger, B, Corkey, B E, Berggren, P O, and Larsson, O
- Abstract
The ATP-dependent potassium (KATP) channel in the pancreatic beta-cell is a complex of two proteins, the pore-forming Kir6.2 and the sulfonylurea receptor type 1 (SUR1). Both subunits are required for functional KATP channels because expression of Kir6.2 alone does not result in measurable currents. However, truncation of the last 26 or 36 amino acids of the C terminus of Kir6.2 enables functional expression of the pore-forming protein in the absence of SUR1. Thus, by using the truncated form of Kir6.2, expressed in the absence and presence of SUR1, it has been shown that the site at which ATP mediates channel inhibition is likely to be situated on Kir6.2. We have now examined the effects of long chain acyl-CoA (LC-CoA) esters on the C-terminally truncated mouse Kir6.2Delta365-390 (Kir6. 2DeltaC26) in inside-out patches isolated from Xenopus laevis oocytes. LC-CoA esters, saturated (C14:0, C16:0) and unsaturated (C18:1), increased Kir6.2DeltaC26 currents, whereas short and medium chain CoA esters (C3:0, C8:0, C12:0) were unable to affect channel activity. The LC-CoA esters were also able to counteract the blocking effect of ATP on Kir6.2DeltaC26. The stimulatory effect of the esters could be explained by the induction of a prolonged open state of Kir6.2DeltaC26. In the presence of the esters, channel open time was increased approximately 3-fold, which is identical to what was obtained in the native mouse KATP channel. Coexpression of SUR1 together with Kir6.2DeltaC26 did not further increase the ability of LC-CoA esters to stimulate channel activity. We conclude that Kir6.2 is the primary target for LC-CoA esters to activate the KATP channel and that the esters are likely to induce a conformational change by a direct interference with the pore-forming subunit, leading to openings of long duration.
- Published
- 1998
19. [Molecular-biological aspects of diabetes mellitus]
- Author
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Walther R, Brandt R, Leibiger I, Leibiger B, Stephan Menzel, Neumer C, and Zühlke H
- Subjects
Animals, Genetically Modified ,Base Sequence ,HLA Antigens ,Molecular Sequence Data ,Diabetes Mellitus ,Animals ,Humans ,Insulin ,Chromosomes, Human, Pair 6 ,Amino Acid Sequence ,DNA ,Polymorphism, Restriction Fragment Length ,Proinsulin
20. 40 EASD Annual Meeting of the European Association for the Study of Diabetes : Munich, Germany, 5-9 September 2004
- Author
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Veitenhansl, M., Stegner, K., Hierl, F-X, Dieterle, C., Feldmeier, H., Gutt, B., Landgraf, R., Garrow, A. P., Vileikyte, L., Findlow, A., Waterman, C., Boulton, A. J. M., Shankhdhar, K., Shankhdhar, L., Shankhdhar, U., Petrova, N. L., Foster, A. V. M., Edmonds, M. E., Ferraresi, R., Caravaggi, C., Giglio, R., Cavaiani, P., Pogliaghi, I., Sommariva, E., Katz, I. A., Harlan, A., Miranda-Palma, B., Prieto-Sanchez, L., Armstrong, D. G., Bowker, J. H., Mizel, M. S., Cernea, S., Wohlgelernter, J., Kidron, M., Modi, P., Raz, I., Arbit, E., Nosek, L., Kapitza, C., Beckett, P., Gelfand, R., Goldberg, M., Heise, T., Testa, M. A., Turner, R. R., Hayes, J. F., Scranton, R. E., Simonson, D. C., Yang, Y-W, Hsu, Y-J, Naujok, O., Francini, F., Jorns, A., Tiedge, M., Lenzen, S., Abdel-Wahab, Y. H. A., Marenah, L., Orr, D. F., Shaw, C., Flatt, P. R., Chokkalingam, K., Mansell, P. I., Clausen, P., Ekbom, P., Damm, P., Feldt-Rasmussen, U., Nielsen, B., Mathiesen, E. R., Feldt-Rasmussen, B., Dewan, S., Da Silva, N., Ternan, P. Mc, Leong, K. S., Wilding, J. P. H., Asatiani, N., Kurashvili, R., Dundua, M., Shelestova, E., Pagava, K., Ramazashvili, M., Hod, M., Smirnov, S., Petersen, J. L. A., Justesen, T. I., Ringholm Nielsen, L., Muller, C., Hojlund, K., Wensaas, A., Kase, E. T., Aas, V., Rustan, A. C., Thoresen, G. H., Levin, K., Beck-Nielsen, H., Gaster, M., Im, S-S, Kang, S-Y, Kim, S-Y, Ahn, Y-H, Lihn, A. S., Schmoll, D., Werner, T., Kienitz, A., Meyer, M., Barthel, A., Ailett, F., Sutherland, C., Walther, R., Grempler, R., Sasson, S., Reich, R., Tenenbaum, T., Alpert, E., Anfossi, G., Russo, I., Traversa, M., Massucco, P., Mattiello, L., Doronzo, G., Trovati, M., Lally, S., Tan, C. Y., Owens, D., Tomkin, G. H., Porchay, I., Pean, F., Bellili, N., Betoulle, D., Balkau, B., Tichet, J., Marre, M., Fumeron, F., Group D.E.S.I.R., Chatellier, G., Alhenc-Gelas, F., Diabhycar, Study Group, Nichols, G. A., Brown, J. B., Hayes, R. P., Bowman, L., Drexel, H., Saely, C. H., Marte, T., Benzer, W., Langer, P., Hoefle, G., Moll, W., Aczel, S., Karagiannis, E., Lubben, G., Urquhart, R., Edwards, G., Bruce, S., Howlett, H. S. C., Cugnardey, N., Turner, K. C., Park, J-S, Fiedorek, F. T., Avogaro, A., Gallo, A., Pinton, P., Rizzuto, R., Murphy, E., Ceolotto, G., Caterson, I., Guy-Grand, B., Hill, J., Barone, M., Aiello, A., Allochis, G., Borzi, V., Cannata, F., Caronna, S., D Avanzo, A., Elli, R., Formoso, G., Paroli, A., Scardapane, R., Sorichetti, P., Tatti, P., Viviani, G., Santeusanio, F., Italian Repaglinide Study Group, Manzella, D., Grella, R., Abbatecola, A. M., Paolisso, G., Sondergaard, L. G., Monster, T. B. M., Johnsen, S. P., Olsen, M. L., Mclaughlin, J. K., Sorensen, H. T., Lervang, H. H., Rungby, J., Lyssenko, V., Fredriksson, J., Almgren, P., Anevski, D., Orho-Melander, M., Sjogren, M., Tuomi, T., Groop, L., Jaziri, R., Aubert, R., Tuomilehto, J., Hu, G., Jousilahti, P., Peltonen, M., Lindstrom, J., Laina, A., Alevizaki, M., Philippou, G., Souvatzoglou, A., Anastasiou, E., Alba, S., Metcalf, B. S., Voss, L. D., Jeffery, A. N., Wilkin, T. J., Gluimer, C., Colagiuri, S., Vistisen, D., Borch-Johnsen, K., Haynes, A., Bower, C., Bulsara, M. K., Jones, T. W., Davis, E. A., Mortensen, H. B., Hougaard, P., Holl, R., Swift, P., Pociot, F., Knip, M., Hansen, L., Szadkowska, A., Pietrzak, I., Zmyslowska, A., Wyka, K., Bodalski, J., Holl, R. W., Swift, R., Hougaard, R., Gerstl, E-M, Engelsberger, I., Rabl, W., Rosenbauer, J., Grobe, H., Hofer, S. E., Krause, U., DPV-Wiss-Study Group, Dabelea, D., Morgan, T., Pettitt, D. J., Dolan, L., Mayer-Davis, E. J., Pihoker, C., Hillier, T. A., Imperatore, G., Ruggiero, A., Hamman, R. E., Stylianou, A., Tentolouris, N., Perrea, D., Tselepis, A. D., Lourida, E., Kitsou, E., Katsilambros, N., Vedovato, M., Dodesini, A. R., Lepore, G., Tiengo, A., Trevisan, R., Penno, G., Miccoli, R., Pucci, L., Lucchesi, D., Bandinelli, S., Fotino, C., Triscornia, S., Baldassari, E., Del Prato, S., Reboldi, P., Santeusanio, E., Fuller, J., Langham, R. G., Gow, R. M., Zhang, Y., Kelly, D. J., Christensen, P. K., Parving, H-H, Gilbert, R. E., Chibalin, A. V., Zhong, Z., Kotova, O., Davidescu, A., Ehren, I., Ekberg, K., Wahren, J., Wassef, L., Buckley, A. J., Rooney, K. B., Briody, J., Thompson, M., Ozanne, S. E., Thompson, C. H., Chamson-Reig, A., Summers, K., Arany, E. J. R., Hill, D. J., Solerte, S. B., Gazzaruso, C., Locatelli, E., Precerutti, S., Schifino, N., Ferrari, E., Fioravanti, M., Phenekos, C. V., Ginis, A., Fragaki, I., Chalkiadaki, M., Tzioras, C., Powell, L. A., Mcguire, G. M., Jewhurst, V., Trimble, E. R., Rasmussen, B. M., Vessby, B., Uusitupa, M., Berglund, L., Pedersen, E., Riccardi, G., Rivellese, A. A., Tapsell, L., Hermansen, K., Kanwu, Study Group, Da Silva Xavier, G., Rutter, J., Rutter, G. A., Briaud, I. M., Lingohr, M. K., Dickson, L. M., Mccuaig, J. R., Lawrence, J. C., Rhodes, C. J., Wikstrom, J. D., Katzman, S. M., Shirihai, O. S., Yang, J., Deng, S., Wang, X., Hessner, M. J., Wu, J., Wong, R. K., Sukumvanich, S., Markman, J. F., Naji, A., Wolf, B. A., Gao, Z., Rubi, B., Del Arco, A., Satrustegui, J., Maechler, P., Del Guerra, S., Lupi, R., Bugliani, M., Sbrana, S., Torri, S., Boggi, U., Vistoli, F., Mosca, F., Marchetti, P., Rennings, A. J. M., Smits, P., Stewart, M. W., Tack, C. J. J., Li, L., Nystrom, T., Gutniak, M., Ahren, B., Holst, J., Sjoholm, A., Gomes, M. B., Cailleaux, S., Tibirica, E., Albertini, J-P, Chen, H., Mather, R., Valensi, P. E., Chisalita, S. I., Arnqvist, H. J., Kraenkel, N., Adams, V., Linke, A., Gielen, S., Schuler, G., Humbrecht, R., Cipollone, F., Iezzi, A., Fazia, M., Pini, B., Cucurullo, C., Cesare, D., Schmidt, A. M., Mazurek, T., Zang, L. F., Mannion, J., Diehl, J., Martin, J., Martella, A., Zalewski, A., Shi, Y., Otter, W., Winter, M., Doering, W., Standi, E., Schnell, O., Kragelund, C., Kober, L., Faber, J., Hildebrandt, P., Steffensen, R., Pankowska, E., Szypowska, A., Lipka, M., Herwig, J., Scholl-Schilling, G., Bohles, H., Robertson, K. J., Schonle, E., Gucev, Z., Mordhorst, L., Tamer, S. C., Gall, M-A, Ludvigsson, J., Hoogma, R. P. L., Hammond, P. J., Gomis, R., Kerr, D., Bruttomesso, D., Bouter, P., Wiefels, K. J., La Calle, H., Schweitzer, D. H., Pfohl, M., Torlone, E., Krinelke, L. G., 205-Nations Study Group, Conget, I., Storms, F., Rodriguez, J., Leperlier, C., Davies, M., At Lantus, Study Group, Peter, R., Luzio, S. D., Dunseath, G., Miles, A., Hare, B., Backx, K., Pauvaday, V., Owens, D. R., Caselli, A., Marfia, G. A., Battista, C., Veves, A., Spallone, V., Uccioli, L., Gonzalez, J. S., Peyrot, M. F., Rubin, R. R., Leventhal, H., Scheffler, N., Ulbrecht, J. S., Cavanagh, P. R., Boulton, A. J., Perrin, N. A., Oglesby, A., Bastyr, E. J., Ziegler, D., Siekierka-Kleiser, E., Meyer, B., Schweers, M., Selvarajah, D., Wilkinson, I. D., Emery, C. J., Shaw, P. J., Griffiths, P. D., Tesfaye, S., Obrosova, I. G., Arezzo, J., Phillips, K., Fidarestat Study Group, Gribble, F. M., Williams, L., Reimann, F., Iakoubov, R., Whiteside, C., Brubaker, P. L., Acitores, A., Gonzalez, N., Sancho, V., Valverde, I., Villanueva-Penacarrillo, M. L., Martin-Duce, A., Trigo, M. V., Arnes, L., Burkart, V., Ichino, N., Ohashi, A., Klein, B. S., Paxian, S., Schmid, R., Karlsen, A. E., Heding, P. E., Frobose, H., Ronn, S. G., Kruhoffer, M., Orntoft, T. F., Nerup, J., Mandrup-Poulsen, T., Billestrup, N., Cardozo, A. K., Ortis, F., Feng, Y-M, Rasschaert, J., Eylen, F., Storling, J., Herchuelz, A., Eizirik, D. L., Wang, H., Kouri, G., Wollheim, C. B., Ribaux, P., Hammar, E., Parnaud, G., Rouiller, D., Bosco, D., Halban, P., Midthjell, K., Carlsson, S., Grill, V., Lau, C., Farch, K., Glumer, C., Tetens, I., Jorgensen, T., Tillin, T., Forouhi, N., Mckeigue, P., Chaturvedi, N., Zethelius, B., Hales, C. N., Berne, C., Coleman, R. L., Stevens, R. J., Holman, R. R., Christensen, J. O., Sandbak, A., Lauritzen, T., Irwin, N., Gault, V. A., Green, B. D., Harriott, P., O Harte, F. P. M., Bouman, S. D., Urso, B., Brand, C. L., Rolin, B., Ribel, U., Schaffer, L., Maggs, D. G., Ceriello, A., Frias, J. P., Wang, Y., Ruggles, J. A., Kolterman, O. G., Piconi, L., Weyer, C., Want, L. L., Ratner, R. E., Uwaifo, G. I., Thornberry, N. A., Eiermann, G., Kim, D., Lankas, G., Leiting, B., Li, Z., Lyons, K., Petrov, A., Sinha Roy, R., Woods, A., Woods, J., Zhang, B. B., Fisher, M., Moller, D. E., Weber, A. E., Dreyer, M., Bellin, C., Schmitz, V., Roesen, R., Nescheret, A. P., Bose, A. K., Mocanu, M. M., Carr, R. D., Yellon, D. M., Manolopoulos, K., Born, S., Wagner, A., Jeziorska, M., Ben Drief, A., Bashir, M., Tomlinson, D., Malik, R. A., Zeymer, U., Schwarzmaier-D Assie, A., Petzinna, D., Chiasson, J-L, Stratton, I. M., Af Bjorkesten, C-G, Fagerudd, J., Rosengard-Barlund, M., Forsblom, C., Pettersson-Fernholm, K., Waden, J., Saraheimo, M., Ronnback, M., Thorn, L., Groop, P-H, Mollsten, A., Svensson, M., Kockum, I., Rudberg, S., Brismar, K., Dahlquist, G., Hovind, P., Hansen, T. K., Tarnow, L., Thiel, S., Jensen, B. R., Flyvbjerg, A., Kankova, K., Hertlova, M., Krusova, D., Schwenke, S., Ott, J., Thom, S. A. M., Mistry, P., Sjolie, A., Larsen, B., Witt, N., Hughes, A. D., Samira, H. H., Lahiry, S., Howlader, S. R., Parveen, S., Azad Khan, A. K., Clarke, P. M., Gray, A., Stevens, R., Holman, R., Phillips, L., Phillips, P. J., Chittleborough, C., Baldock, K., Taylor, A., North West Adelaide Health Study Team, Davis, W. A., Davis, T. M. E., Knuiman, M. W., Hendrie, D., Worthley, D., Nicolucci, A., Pellegrini, F., Berardis, G., Franciosi, M., Belfiglio, M., Rossi, M. C. E., Sacco, M., Valentini, M., Richardson, C. C., Jones, P., Persaud, S., Hussain, K., Clark, A., Christie, M. R., Gniuli, D., Hribal, M. L., Accili, D., Khan, M., Zervou, S., Cheung, L., Abouna, S., Ifandi, V., Pelengaris, S., Luco, R. F., Ferrer, J., Ma, D., Shield, J. P. H., Dean, W., Leclerc, I., Knauf, C., Burcelin, R., Kelsey, G., Powers, A. C., Shostak, A., Ferrara, N., Poffenberger, G., Jerome, W. G., Brissova, M., Geloneze, S. R., Tambascia, M. A., Pareja, J. C., Chaim, E., Silveira, H. V., Geloneze, B., Ravikumar, B., Carey, P. E., Snaar, J. E., Dheelchand, D., Cook, D. B., Neely, D., Taylor, G., Morris, P. G., Taylor, R., Stears, A. J., Masding, M. G., Wootton, S. A., Sandeman, D. D., Klimes, I., Wein, S., Gasperikova, D., Ukropec, J., Wiernsperger, N., Sebokova, E., Manco, M., Mingrone, G., Granato, L., Greco, A. V., Nanni, G., Castagneto, M., Vidal, H., Calvani, M., Ferrannini, E., Alvarsson, M., Sundkvist, G., Lager, I., Henricsson, M., Berntorp, K., Fernqvist-Forbes, E., Steen, L., Orn, T., Shutler, S., Bianchi-Biscay, M., Rosenstock, J., Sugimoto, D., Strange, P., Stewart, J., Soltes Rak, E., Dailey, G., Kloos, C., Muller, U., Samann, A., Femerling, M., Risse, A., Jecht, M., Haak, T., Garg, R., Lawrence, I. G., Akinsola, M. O., Davies, M. J., Mcnally, P. G., Garber, A. J., Kim, H., Draeger, E., Aydin, L., Sengul, A., Kurklu, A., Ucak, S., Basat, O., Seber, S., Altuntas, Y., Jin, J., Yu, Y., Yu, H., Zhang, X., Mattoo, V., Eckland, D., Widel, M., Duran, S., Fajardo, C., Strand, J., Knight, D., Oakley, D., Tan, M., Sato, A., Nagao, M., Aki, N., Nakagami, T., Iwamoto, Y., Zhou, Z., Li, X., Huang, G., Yan, X., Yang, L., Peng, J., Wang, J., Tan, S., Tang, W., Furnsinn, C., Brunmair, B., Wagner, L., Gras, F., Artwohl, M., Zierhut, B., Waldhausl, W., Shine, B. L., Hopkins, D., Anand, V., Lim, E., Raval, U., Sharp, P., Corder, R., Lipkin, D., Lahiri, A., Bartnik, M., Ryden, L., Ferrari, R., Malmberg, K., Pyorala, K., Simoons, M. L., Standl, E., Soler-Soler, J., Ohrvik, J., Euro Heart Survey Investigators, Bruce, D. G., Starkstein, S. E., Schauer, U. J. W., Astrup, A. S., Pietraszek, L., Nielsen, F. S., Rossing, P., Ali, S., Smidt, U. M., Yokoyama, H., Pavkov, M. E., Knowler, W. C., Bennett, P. H., Nelson, R. G., Lopez-Alba, A., Morcillo, L., Caballero, A., Montoya, L., Jimenez, A., Maceira, B., Lewis, J. B., Ravid, M., Wajman, A., Tadgell, C., Remuzzi, G., Hunsicker, L. G., Wessman, M., Taskinen, M-R, FinnDiane Study Group, Pugliese, G., Amadio, L., Menini, S., Oddi, G., Ricci, C., Iacobini, C., Pricci, F., Sorcini, M., Pesce, C., Migliaccio, E., Giorgio, M., Pelicci, P., Di Mario, U., Lassila, M., Jandeleit-Dahm, K., Seah, K. K., Calkin, A. C., Allen, T. J., Cooper, M. E., Lopes Faria, J. M., Cavakcanti, T. C., Silva, K. C., Ferrari, A. L., Lopes Faria, J. B., Cellek, S., Foxwell, N. A., Cotter, M. A., Cameron, N. E., Tennagels, N., Jordan, H., Stahl, P., Voss, M. D., Welte, S., Werner, U., Lehmann, R., Moeschel, K., Baumgartner, F., Oeckinghaus, A., Beck, A., Weigert, C., Hennige, A., Schleicher, E. D., Haring, H. U., Mussig, K., Staiger, H., Haring, H-U, Natalicchio, A., Laviola, L., Tullio, C., Renna, L., Giorgino, R., Giorgino, F., Falasca, M., Maffucci, T., Park, D., Kang, S., Song, J., Lee, D., Lee, Y., Hariharan, N., Kunselman, L., Gu, L., Sasseville, V., Harrity, T., Cheng, P. T. W., Pratley, R. E., Schweizer, A., Mills, D., Kim, T-H, Song, X-L, Poelje, P. D., Potter, S. C., Dang, Q., Fujitaki, J. M., Linemeyer, D. L., Landau, B. R., Erion, M. D., Pankop, M., Golay, A., Despres, J., Sjostrom, L., Lawlor, D. L., Legg, K. T., Ur, E., Houweling, S. T., Kleefstra, N., Meyboom-De Jong, B., Bilo, H. J. G., Schlomer, G. 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21. Positive and negative regulatory elements are involved in transcriptional control of the rat glucokinase gene in the insulin producing cell line HIT M2.2.2
- Author
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Leibiger, I. B., Walther, R., Pett, U., and Leibiger, B.
- Published
- 1994
- Full Text
- View/download PDF
22. Functional analysis of a newly identified TAAT-box of the rat insulin-II gene promoter
- Author
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Leibiger, I. B., Schwarz, T., Leibiger, B., and Walther, R.
- Published
- 1995
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23. CT-Boxes are involved in control of the rat insulin II gene expression
- Author
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Rommel, C., Leibiger, I. B., Leibiger, B., and Walther, R.
- Published
- 1994
- Full Text
- View/download PDF
24. 40(th) EASD Annual Meeting of the European Association for the Study of Diabetes : Munich, Germany, 5-9 September 2004
- Author
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S. Artigas, A V Dreval, Mark I. McCarthy, C Watson, Peter H. Bennett, M Quint, Y Ikeda, E Alpert, F Schiele, H Sekihara, Erik Gylfe, P Lowe, J Kuhlmann, Alain Golay, V Longo, Shahidul Alam Khan Akm., L G Mantovani, M Zawodniak-Szalapska, G Winkler, T Harrity, L Virág, U Johne, Kuo S-W., Linda C Tapsell, J Rodriguez, Michel Komajda, K Kankova, Carole A. Cull, M Sporna, E Estilles, U Ribel, M C Spruce, E Buzzigoli, T Prazak, J K McLaughlin, M K Lingohr, M Lim, F Calara, A Siebenhofer, G Meregalli, Roberto Anichini, A D Baron, R Kurashvili, P C Butler, G I Fantus, T. E. De Gooyer, Park Y-M., R. Walther, S Heinrich, Agnieszka Zawiejska, S Mukherjee, Nikolaos Papanas, G Wong, Ian D. Caterson, David M. Maahs, Shuichi Kaneko, Alexandra E. Butler, Francisco Javier Ampudia-Blasco, O N Kong, Attali J-R., C A Hedman, K Oshinyemi, Nicolle Müller, I C Cranston, N Okumus, M V Vlaiculescu, Balasubramanian Ravikumar, W W Cheatham, K Mukasa, K B Biswas, Annunziata Lapolla, Phil McEwan, G Mader, Gilles Chassot, Dragi Anevski, Werner A. Scherbaum, M Donath, C Hesselmann, R A Gandhi, David E. Moller, Ezio Bonifacio, C Garcia, V Ifandi, P Hornnes, Nieuwenhoven Fav., C Puech, S Pérez-Del-Pulgar, Kim S-R., G Hines, C Rubio Terrés, Michael Gaster, N. Hosszufalusi, A Scholze, Andrew A. Young, Stavros Liatis, F Hariri, S Tan, Paul Valensi, Allan E. Karlsen, J Kim, E. Moberg, J Kaiser, L Berman, G Nelson, A Altkrüger, P Kothare, D B Cook, S Doran, G. van Dijk, Shahnaz Shahinfar, Kim C-S., P Stahl, M Manousaki, S Sigrist, S K Lim, M. P. Stern, A Guberti, C Rezzani, J McKenney, Karl Thomaseth, Sofia Carlsson, M Julia, R Brillante, I Rubesova, T Darkow, E Matsumoto, Wendy M. 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Dell'Antonio, G, Maestroni, A, Ruggieri, D, Luzi, L, Piemonti, L, Zerbini, G, Anafaroglu, I, Tutuncu, N, Sultana, M, Siddiqua, N, Iwasaki, T, Nakajima, A, Yoneda, M, Mukasa, K, Tanaka, S, and Sekihara, H
- Subjects
0303 health sciences ,medicine.medical_specialty ,business.industry ,EASD ,Endocrinology, Diabetes and Metabolism ,Human physiology ,medicine.disease ,03 medical and health sciences ,0302 clinical medicine ,Diabetes mellitus ,Family medicine ,Internal Medicine ,Medicine ,business ,030217 neurology & neurosurgery ,030304 developmental biology - Published
- 2004
25. Local Dexamethasone Administration Delays Allogeneic Islet Graft Rejection in the Anterior Chamber of the Eye of Non-Human Primates.
- Author
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Tun SBB, Chua M, Tan GSW, Leibiger I, Ali Y, Barathi VA, and Berggren PO
- Subjects
- Animals, Anterior Chamber, Dexamethasone pharmacology, Dexamethasone therapeutic use, Graft Rejection, Graft Survival, Mice, Primates, Diabetes Mellitus, Hematopoietic Stem Cell Transplantation, Islets of Langerhans Transplantation methods
- Abstract
Pancreatic islet transplantation into the anterior chamber of the eye (ACE) has been shown to improve glycemic control and metabolic parameters of diabetes in both murine and primate models. This novel transplantation site also allows the delivery of therapeutic agents, such as immunosuppressive drugs, locally to prevent islet graft rejection and circumvent unwanted systemic side effects. Local intravitreal administration of micronized dexamethasone implant was performed prior to allogeneic islet transplantation into the ACEs of non-human primates. Two study groups were observed namely allogeneic graft without immunosuppression (n = 4 eyes) and allogeneic graft with local immunosuppression (n = 8 eyes). Survival of islet grafts and dexamethasone concentration in the ACE were assessed in parallel for 24 weeks. Allogeneic islet grafts with local dexamethasone treatment showed significantly better survival than those with no immunosuppression (median survival time- 15 weeks vs 3 weeks, log-rank test p<0.0001). Around 73% of the grafts still survived at week 10 with a single local dexamethasone implant, where the control group showed no graft survival. Dexamethasone treated islet grafts revealed a good functional response to high glucose stimulation despite there was a transient suppression of insulin secretion from week 8 to 12. Our findings show a significant improvement of allografts survival in the ACE with local dexamethasone treatment. These results highlight the feasibility of local administration of pharmacological compounds in the ACE to improve islet graft survival and function. By eliminating the need for systemic immunosuppression, these findings may impact clinical islet transplantation in the treatment of diabetes, and the ACE may serve as a novel therapeutic islet transplantation site with high potential for local pharmacological intervention.
- Published
- 2022
- Full Text
- View/download PDF
26. Author Correction: Structural basis for delta cell paracrine regulation in pancreatic islets.
- Author
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E Drigo RA, Jacob S, García-Prieto CF, Zheng X, Fukuda M, Nhu HTT, Stelmashenko O, Peçanha FLM, Rodriguez-Diaz R, Bushong E, Deerinck T, Phan S, Ali Y, Leibiger I, Chua M, Boudier T, Song SH, Graf M, Augustine GJ, Ellisman MH, and Berggren PO
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
- Published
- 2019
- Full Text
- View/download PDF
27. Structural basis for delta cell paracrine regulation in pancreatic islets.
- Author
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Arrojo E Drigo R, Jacob S, García-Prieto CF, Zheng X, Fukuda M, Nhu HTT, Stelmashenko O, Peçanha FLM, Rodriguez-Diaz R, Bushong E, Deerinck T, Phan S, Ali Y, Leibiger I, Chua M, Boudier T, Song SH, Graf M, Augustine GJ, Ellisman MH, and Berggren PO
- Subjects
- Animals, Blood Glucose metabolism, Humans, Insulin-Like Growth Factor I metabolism, Insulin-Secreting Cells cytology, Insulin-Secreting Cells metabolism, Insulin-Secreting Cells ultrastructure, Intravital Microscopy, Islets of Langerhans cytology, Islets of Langerhans metabolism, Islets of Langerhans pathology, Mice, Mice, Transgenic, Microscopy, Electron, Optical Imaging, Optogenetics, Prediabetic State metabolism, Pseudopodia metabolism, Somatostatin-Secreting Cells cytology, Somatostatin-Secreting Cells metabolism, Vascular Endothelial Growth Factor A metabolism, Islets of Langerhans ultrastructure, Paracrine Communication, Prediabetic State pathology, Pseudopodia ultrastructure, Somatostatin-Secreting Cells ultrastructure
- Abstract
Little is known about the role of islet delta cells in regulating blood glucose homeostasis in vivo. Delta cells are important paracrine regulators of beta cell and alpha cell secretory activity, however the structural basis underlying this regulation has yet to be determined. Most delta cells are elongated and have a well-defined cell soma and a filopodia-like structure. Using in vivo optogenetics and high-speed Ca
2+ imaging, we show that these filopodia are dynamic structures that contain a secretory machinery, enabling the delta cell to reach a large number of beta cells within the islet. This provides for efficient regulation of beta cell activity and is modulated by endogenous IGF-1/VEGF-A signaling. In pre-diabetes, delta cells undergo morphological changes that may be a compensation to maintain paracrine regulation of the beta cell. Our data provides an integrated picture of how delta cells can modulate beta cell activity under physiological conditions.- Published
- 2019
- Full Text
- View/download PDF
28. Mechanistic understanding of insulin receptor modulation: Implications for the development of anti-diabetic drugs.
- Author
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Yunn NO, Kim J, Kim Y, Leibiger I, Berggren PO, and Ryu SH
- Subjects
- Animals, Diabetes Mellitus drug therapy, Humans, Hypoglycemic Agents therapeutic use, Insulin pharmacology, Insulin therapeutic use, Diabetes Mellitus metabolism, Hypoglycemic Agents pharmacology, Receptor, Insulin metabolism
- Abstract
The insulin receptor is an important regulator of metabolic processes in the body, and in particular of glucose homeostasis, including glucose uptake into peripheral tissues. Thus, insulin administration is an effective treatment for diabetes, which is characterized by chronic elevation of blood glucose. However, insulin is not only a metabolic regulator, but also functions as a growth hormone. Accordingly, studies of long-term insulin administration and of the hyperinsulinemia associated with type 2 diabetes have raised concerns about possible increases in the risks of cancer and atherosclerosis, due to excessive stimulation of cell proliferation. Interestingly, some insulin receptor ligands that have been developed based on a peptide, an antibody, and an aptamer selectively have metabolic effects exerted through the insulin receptor but do not cause significant cellular proliferation. Although these ligands therefore have potential as anti-diabetic drugs for advanced diabetes care, the mechanism whereby they specifically activate the insulin receptor is still unclear. Recently, studies of the structure of the insulin receptor have progressed considerably, and have provided further mechanistic understanding of insulin receptor activation. Based on this progress, we propose a mechanistic model of this specificity and discuss the potential for the development of novel anti-diabetic drugs that would not have the adverse effects caused by excessive mitogenic action., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2018
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29. Pancreatic Islet Blood Flow Dynamics in Primates.
- Author
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Diez JA, Arrojo E Drigo R, Zheng X, Stelmashenko OV, Chua M, Rodriguez-Diaz R, Fukuda M, Köhler M, Leibiger I, Tun SBB, Ali Y, Augustine GJ, Barathi VA, and Berggren PO
- Subjects
- Animals, Blood Glucose metabolism, Capillaries physiology, Cells, Cultured, Hypoglycemic Agents pharmacology, Islets of Langerhans drug effects, Liraglutide pharmacology, Macaca fascicularis, Male, Regional Blood Flow, Vasoconstriction, Blood Flow Velocity, Islets of Langerhans blood supply
- Abstract
Blood flow regulation in pancreatic islets is critical for function but poorly understood. Here, we establish an in vivo imaging platform in a non-human primate where islets transplanted autologously into the anterior chamber of the eye are monitored non-invasively and longitudinally at single-cell resolution. Engrafted islets were vascularized and innervated and maintained the cytoarchitecture of in situ islets in the pancreas. Blood flow velocity in the engrafted islets was not affected by increasing blood glucose levels and/or the GLP-1R agonist liraglutide. However, islet blood flow was dynamic in nature and fluctuated in various capillaries. This was associated with vasoconstriction events resembling a sphincter-like action, most likely regulated by adrenergic signaling. These observations suggest a mechanism in primate islets that diverts blood flow to cell regions with higher metabolic demand. The described imaging technology applied in non-human primate islets may contribute to a better understanding of human islet pathophysiology., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
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30. The paired basic amino acid-cleaving enzyme 4 (PACE4) is involved in the maturation of insulin receptor isoform B: an opportunity to reduce the specific insulin receptor-dependent effects of insulin-like growth factor 2 (IGF2).
- Author
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Kara I, Poggi M, Bonardo B, Govers R, Landrier JF, Tian S, Leibiger I, Day R, Creemers JW, and Peiretti F
- Subjects
- 3T3-L1 Cells, Animals, Cell Proliferation, Furin genetics, Furin metabolism, HEK293 Cells, HeLa Cells, Humans, Immunoblotting, Mice, Proprotein Convertases genetics, Real-Time Polymerase Chain Reaction, Receptor, Insulin genetics, Serine Endopeptidases genetics, Insulin-Like Growth Factor II metabolism, Proprotein Convertases metabolism, Receptor, Insulin metabolism, Serine Endopeptidases metabolism
- Abstract
Gaining the full activity of the insulin receptor (IR) requires the proteolytic cleavage of its proform by intra-Golgi furin-like activity. In mammalian cells, IR is expressed as two isoforms (IRB and IRA) that are responsible for insulin action. However, only IRA transmits the growth-promoting and mitogenic effects of insulin-like growth factor 2. Here we demonstrate that the two IR isoforms are similarly cleaved by furin, but when this furin-dependent maturation is inefficient, IR proforms move to the cell surface where the proprotein convertase PACE4 selectively supports IRB maturation. Therefore, in situations of impaired furin activity, the proteolytic maturation of IRB is greater than that of IRA, and accordingly, the amount of phosphorylated IRB is also greater than that of IRA. We highlight the ability of a particular proprotein convertase inhibitor to effectively reduce the maturation of IRA and its associated mitogenic signaling without altering the signals emanating from IRB. In conclusion, the selective PACE4-dependent maturation of IRB occurs when furin activity is reduced; accordingly, the pharmacological inhibition of furin reduces IRA maturation and its mitogenic potential without altering the insulin effects., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
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31. Noninvasive in vivo model demonstrating the effects of autonomic innervation on pancreatic islet function.
- Author
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Rodriguez-Diaz R, Speier S, Molano RD, Formoso A, Gans I, Abdulreda MH, Cabrera O, Molina J, Fachado A, Ricordi C, Leibiger I, Pileggi A, Berggren PO, and Caicedo A
- Subjects
- Animals, Green Fluorescent Proteins metabolism, Iris innervation, Iris physiology, Islets of Langerhans Transplantation, Mice, Mice, 129 Strain, Mice, Inbred C57BL, Mice, Transgenic, Nerve Fibers, Autonomic Nervous System physiology, Eye innervation, Islets of Langerhans physiology, Models, Biological
- Abstract
The autonomic nervous system is thought to modulate blood glucose homeostasis by regulating endocrine cell activity in the pancreatic islets of Langerhans. The role of islet innervation, however, has remained elusive because the direct effects of autonomic nervous input on islet cell physiology cannot be studied in the pancreas. Here, we used an in vivo model to study the role of islet nervous input in glucose homeostasis. We transplanted islets into the anterior chamber of the eye and found that islet grafts became densely innervated by the rich parasympathetic and sympathetic nervous supply of the iris. Parasympathetic innervation was imaged intravitally by using transgenic mice expressing GFP in cholinergic axons. To manipulate selectively the islet nervous input, we increased the ambient illumination to increase the parasympathetic input to the islet grafts via the pupillary light reflex. This reduced fasting glycemia and improved glucose tolerance. These effects could be blocked by topical application of the muscarinic antagonist atropine to the eye, indicating that local cholinergic innervation had a direct effect on islet function in vivo. By using this approach, we found that parasympathetic innervation influences islet function in C57BL/6 mice but not in 129X1 mice, which reflected differences in innervation densities and may explain major strain differences in glucose homeostasis. This study directly demonstrates that autonomic axons innervating the islet modulate glucose homeostasis.
- Published
- 2012
- Full Text
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32. Dynamin-mediated Nephrin phosphorylation regulates glucose-stimulated insulin release in pancreatic beta cells.
- Author
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Jeon J, Leibiger I, Moede T, Walter B, Faul C, Maiguel D, Villarreal R, Guzman J, Berggren PO, Mundel P, Ricordi C, Merscher-Gomez S, and Fornoni A
- Subjects
- Amino Acid Substitution, Dynamins genetics, Gene Silencing, Glucose pharmacology, HEK293 Cells, Humans, Insulin Secretion, Insulin-Secreting Cells cytology, Membrane Proteins genetics, Mutation, Missense, Phosphorylation drug effects, Phosphorylation physiology, Sweetening Agents metabolism, Sweetening Agents pharmacology, Vanadates pharmacology, Dynamins metabolism, Glucose metabolism, Insulin metabolism, Insulin-Secreting Cells metabolism, Membrane Proteins metabolism, Proteolysis
- Abstract
We have previously demonstrated a role for Nephrin in glucose stimulated insulin release (GSIR). We now hypothesize that Nephrin phosphorylation is required for GSIR and that Dynamin influences Nephrin phosphorylation and function. MIN6-C3 Nephrin-deficient pancreatic beta cells and human islets were transfected with WT-Nephrin or with a mutant Nephrin in which the tyrosine residues responsible for SH2 domain binding were substituted with phenylalanine (3YF-Nephrin). GSIR and live images of Nephrin and vesicle trafficking were studied. Immunoprecipitation experiments and overexpression of WT-Dynamin or dominant negative Dynamin mutant (K44A-Dynamin) in WT-Nephrin, 3YF-Nephrin, or Nephrin siRNA-transfected cells were utilized to study Nephrin-Dynamin interaction. In contrast to WT-Nephrin or to single tyrosine mutants, 3YF-Nephrin did not positively affect GSIR and led to impaired cell-cell contacts and vesicle trafficking. K44A-Dynamin prevented the effect of Nephrin on GSIR in the absence of protein-protein interaction between Nephrin and Dynamin. Nephrin gene silencing abolished the positive effects of WT-Dynamin on GSIR. The effects of protamine sulfate and vanadate on Nephrin phosphorylation and GSIR were studied in MIN6 cells and human islets. WT-Nephrin phosphorylation after glucose occurred at Tyr-1176/1193 and resulted in improved GSIR. On the contrary, protamine sulfate-induced phosphorylation at Tyr-1176/1193/1217 was associated with Nephrin degradation and impaired GSIR. Vanadate, which prevented Nephrin dephosphorylation after glucose stimulation, improved GSIR in human islets and MIN6 cells. In conclusion, Dynamin-dependent Nephrin phosphorylation occurs in response to glucose and is necessary for Nephrin-mediated augmentation of GSIR. Pharmacological modulation of Nephrin phosphorylation may thus facilitate pancreatic beta cell function.
- Published
- 2012
- Full Text
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33. High-resolution, noninvasive longitudinal live imaging of immune responses.
- Author
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Abdulreda MH, Faleo G, Molano RD, Lopez-Cabezas M, Molina J, Tan Y, Echeverria OA, Zahr-Akrawi E, Rodriguez-Diaz R, Edlund PK, Leibiger I, Bayer AL, Perez V, Ricordi C, Caicedo A, Pileggi A, and Berggren PO
- Subjects
- Amides pharmacology, Animals, Anterior Chamber metabolism, Anterior Chamber surgery, CCR5 Receptor Antagonists, Chemokines pharmacology, Diabetes Mellitus, Experimental therapy, Enzyme-Linked Immunosorbent Assay, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Interferon-gamma metabolism, Interleukin-2 metabolism, Islets of Langerhans metabolism, Islets of Langerhans Transplantation methods, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Microscopy, Video methods, Quaternary Ammonium Compounds pharmacology, Receptors, CCR5 metabolism, Single-Cell Analysis methods, T-Lymphocytes drug effects, T-Lymphocytes metabolism, Time Factors, Anterior Chamber cytology, Islets of Langerhans cytology, Microscopy, Confocal methods, T-Lymphocytes cytology
- Abstract
Intravital imaging emerged as an indispensible tool in biological research, and a variety of imaging techniques have been developed to noninvasively monitor tissues in vivo. However, most of the current techniques lack the resolution to study events at the single-cell level. Although intravital multiphoton microscopy has addressed this limitation, the need for repeated noninvasive access to the same tissue in longitudinal in vivo studies remains largely unmet. We now report on a previously unexplored approach to study immune responses after transplantation of pancreatic islets into the anterior chamber of the mouse eye. This approach enabled (i) longitudinal, noninvasive imaging of transplanted tissues in vivo; (ii) in vivo cytolabeling to assess cellular phenotype and viability in situ; (iii) local intervention by topical application or intraocular injection; and (iv) real-time tracking of infiltrating immune cells in the target tissue.
- Published
- 2011
- Full Text
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34. CREB3L2-PPARgamma fusion mutation identifies a thyroid signaling pathway regulated by intramembrane proteolysis.
- Author
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Lui WO, Zeng L, Rehrmann V, Deshpande S, Tretiakova M, Kaplan EL, Leibiger I, Leibiger B, Enberg U, Höög A, Larsson C, and Kroll TG
- Subjects
- Base Sequence, Cell Membrane metabolism, Cell Proliferation, DNA Primers, Humans, Hydrolysis, In Situ Hybridization, Fluorescence, Molecular Sequence Data, Reverse Transcriptase Polymerase Chain Reaction, Thyroid Neoplasms pathology, Transcription, Genetic, Basic-Leucine Zipper Transcription Factors genetics, Mutation, PPAR gamma genetics, Signal Transduction, Thyroid Neoplasms metabolism
- Abstract
The discovery of gene fusion mutations, particularly in leukemia, has consistently identified new cancer pathways and led to molecular diagnostic assays and molecular-targeted chemotherapies for cancer patients. Here, we report our discovery of a novel CREB3L2-PPARgamma fusion mutation in thyroid carcinoma with t(3;7)(p25;q34), showing that a family of somatic PPARgamma fusion mutations exist in thyroid cancer. The CREB3L2-PPARgamma fusion encodes a CREB3L2-PPARgamma fusion protein that is composed of the transactivation domain of CREB3L2 and all functional domains of PPARgamma1. CREB3L2-PPARgamma was detected in <3% of thyroid follicular carcinomas. Engineered overexpression of CREB3L2-PPARgamma induced proliferation by 40% to 45% in primary human thyroid cells, consistent with a dominant oncogenic mechanism. Wild-type CREB3L2 was expressed in the thyroid as a bZIP transcription factor with a transmembrane domain that has flanking S1P and S2P proteolytic cleavage sites. Native CREB3L2 was cleaved to nuclear CREB3L2 by regulated intramembrane proteolysis in normal thyroid cells that expressed the S1P and S2P proteases. Nuclear CREB3L2 stimulated transcription 8-fold from the EVX1 cyclic AMP (cAMP) response element in the absence of cAMP, whereas CREB3L2-PPARgamma inhibited transcription 6-fold from EVX1 in the same experiments. CREB3L2-PPARgamma also inhibited 4-fold the expression of thyroglobulin, a native cAMP-responsive gene, in primary thyroid cells treated with thyroid-stimulating hormone. Our findings identify a novel CREB3L2-PPARgamma gene fusion mutation in thyroid carcinoma and reveal a thyroid signaling pathway that is regulated by intramembrane proteolysis and disrupted in cancer.
- Published
- 2008
- Full Text
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35. Real-time monitoring of apoptosis by caspase-3-like protease induced FRET reduction triggered by amyloid aggregation.
- Author
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Paulsson JF, Schultz SW, Köhler M, Leibiger I, Berggren PO, and Westermark GT
- Subjects
- Amino Acid Sequence, Animals, Cell Line, Tumor, Cells, Cultured, Diabetes Mellitus, Type 2 metabolism, Fluorescence Resonance Energy Transfer, Insulin metabolism, Insulin-Secreting Cells metabolism, Insulin-Secreting Cells pathology, Islet Amyloid Polypeptide, Mice, Mice, Transgenic, Recombinant Proteins metabolism, Transfection, Amyloid metabolism, Apoptosis, Caspase 3 metabolism, Insulinoma metabolism, Insulinoma pathology, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms pathology
- Abstract
Amyloid formation is cytotoxic and can activate the caspase cascade. Here, we monitor caspase-3-like activity as reduction of fluorescence resonance energy transfer (FRET) using the contstruct pFRET2-DEVD containing enhanced cyan fluorescent protin (EYFP) linked by the caspase-3 specific cleavage site residues DEVD. Beta-TC-6 cells were transfected, and the fluoorescence was measured at 440 nm excitation and 535 nm (EYFP) and 480 nm (ECFP) emission wavelength. Cells were incubated with recombinant pro lset Amyloid Polypeptide (rec prolAPP) or the processing metabolites of prolAPP; the N-terminal flanking peptide withIAPP (recN+IAPP); IAPP with the C-terminal flanking peptied (recIAPP+C) and lslet Amyloid Polypeptide (recIAPP) . Peptides were added in solubilized from (50 microM) or as performed amyloid-like fibrils, or as a combination of these. FRET was measured and incubation with a mixture of solubilized peptide and performed fibrils resulted in loss of FRET and apoptosis was determined to occure in cells incubated with recproIAPP (49%), recN+IAPP (46%), recIAPP (72%) and recIAPP+C (59%). These results show that proIAPP and the processing intermediates reside the same cell toxic capacity as IAPP, and they can all have a central role in the reduction of beta-cell number in type 2 diabetes.
- Published
- 2008
- Full Text
- View/download PDF
36. Monomeric G-protein, Rhes, is not an imidazoline-regulated protein in pancreatic beta-cells.
- Author
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Sharoyko VV, Zaitseva II, Varsanyi M, Portwood N, Leibiger B, Leibiger I, Berggren PO, Efendić S, and Zaitsev SV
- Subjects
- Animals, Cells, Cultured, Down-Regulation drug effects, GTP-Binding Proteins genetics, Insulin metabolism, Insulin Secretion, Male, Monomeric GTP-Binding Proteins genetics, RNA, Messenger genetics, Rats, Rats, Wistar, GTP-Binding Proteins metabolism, Imidazolines pharmacology, Islets of Langerhans drug effects, Islets of Langerhans metabolism, Monomeric GTP-Binding Proteins metabolism
- Abstract
The monomeric G-protein, Rhes, is a candidate imidazoline-regulated molecule involved in mediating the insulin secretory response to efaroxan [S.L. Chan, L.K. Monks, H. Gao, P. Deaville, N.G. Morgan, Identification of the monomeric G-protein, Rhes, as an efaroxan-regulated protein in the pancreatic beta-cell, Br. J. Pharmacol. 136 (1) (2002) 31-36]. This suggestion was based on observations regarding changes in Rhes mRNA expression in rat islets and pancreatic beta-cells after prolonged culture with efaroxan, leading to desensitization of the insulin response to the compound. To verify this report, we have evaluated the effects of the imidazoline compounds efaroxan and BL11282 on Rhes mRNA expression in isolated rat pancreatic islets maintained in conditions identical to those used by Chan et al. The results demonstrate that desensitization of the insulin response to efaroxan, or to another imidazoline, BL11282, does not change Rhes mRNA expression levels. Transfection of MIN6 cells with plasmids containing Rhes or Rhes-antisense also does not alter efaroxan- or BL11282-induced insulin secretion. Together, these data do not support the hypothesis that Rhes is an imidazoline-regulated protein.
- Published
- 2005
- Full Text
- View/download PDF
37. Apolipoprotein CIII promotes Ca2+-dependent beta cell death in type 1 diabetes.
- Author
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Juntti-Berggren L, Refai E, Appelskog I, Andersson M, Imreh G, Dekki N, Uhles S, Yu L, Griffiths WJ, Zaitsev S, Leibiger I, Yang SN, Olivecrona G, Jörnvall H, and Berggren PO
- Subjects
- Adult, Animals, Apolipoprotein C-III, Calcium Channels, L-Type physiology, Diabetes Mellitus, Type 1 blood, Female, Humans, Male, Mice, Apolipoproteins C physiology, Apoptosis, Calcium physiology, Diabetes Mellitus, Type 1 pathology, Islets of Langerhans pathology
- Abstract
In type 1 diabetes (T1D), there is a specific destruction of the insulin secreting pancreatic beta cell. Although the exact molecular mechanisms underlying beta cell destruction are not known, sera from T1D patients have been shown to promote Ca(2+)-induced apoptosis. We now demonstrate that apolipoprotein CIII (apoCIII) is increased in serum from T1D patients and that this serum factor both induces increased cytoplasmic free intracellular Ca(2+) concentration ([Ca(2+)](i)) and beta cell death. The apoCIII-induced increase in [Ca(2+)](i) reflects an activation of the voltage-gated L-type Ca(2+) channel. Both the effects of T1D sera and apoCIII on the beta cell are abolished in the presence of antibody against apoCIII. Increased serum levels of apoCIII can thus account for the increase in beta cell [Ca(2+)](i) and thereby beta cell apoptosis associated with T1D.
- Published
- 2004
- Full Text
- View/download PDF
38. Interferon alpha-induced apoptosis in tumor cells is mediated through the phosphoinositide 3-kinase/mammalian target of rapamycin signaling pathway.
- Author
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Thyrell L, Hjortsberg L, Arulampalam V, Panaretakis T, Uhles S, Dagnell M, Zhivotovsky B, Leibiger I, Grandér D, and Pokrovskaja K
- Subjects
- Adaptor Proteins, Signal Transducing, Annexin A5 pharmacology, Antibiotics, Antineoplastic pharmacology, Blotting, Western, Carrier Proteins metabolism, Caspases metabolism, Cell Cycle Proteins, Cell Death, Cell Line, Tumor, Cell Nucleus metabolism, Chromones pharmacology, Coloring Agents pharmacology, Enzyme Activation, Enzyme Inhibitors pharmacology, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Green Fluorescent Proteins, Humans, Immunohistochemistry, Interferon-alpha metabolism, Keratinocytes metabolism, Luminescent Proteins metabolism, Membrane Potentials, Mitochondria metabolism, Morpholines pharmacology, Phosphoproteins metabolism, Phosphorylation, Recombinant Fusion Proteins metabolism, Ribosomal Protein S6 Kinases, 70-kDa metabolism, Sirolimus pharmacology, TOR Serine-Threonine Kinases, Transfection, Apoptosis, Interferon-alpha pharmacology, Phosphatidylinositol 3-Kinases metabolism, Protein Kinases metabolism, Signal Transduction
- Abstract
Interferon (IFN) alpha induces a caspase-dependent apoptosis that is associated with activation of the proapoptotic Bak and Bax, loss of mitochondrial membrane potential, and release of cytochrome c. In addition to the onset of the classical Jak-STAT pathway, IFNalpha also induced phosphoinositide 3-kinase (PI3K) activity. Pharmacological inhibition of PI3K activity by Ly294002 disrupted IFN-induced apoptosis upstream of mitochondria. Inhibition of mTOR by rapamycin or by overexpression of a kinase dead mutant of mTOR, efficiently blocked IFNalpha-induced apoptosis. A PI3K and mTOR-dependent phosphorylation of p70S6 kinase and 4E-BP1 repressor was induced by IFNalpha treatment of cells and was strongly inhibited by Ly294002 or rapamycin. The activation of Jak-STAT signaling upon IFNalpha stimulation was not affected by abrogating PI3K/mTOR pathway. Neither was the expression of several IFNalpha target genes affected, nor the ability of IFNalpha to protect against virus-induced cell death affected by inhibition of the PI3K/mTOR pathway. These data demonstrate that an intact PI3K/mTOR pathway is necessary for the ability of IFNalpha to induce apoptosis, whereas activation of the Jak-STAT pathway alone appears to be insufficient for this specific IFNalpha-induced effect.
- Published
- 2004
- Full Text
- View/download PDF
39. Characterization of a putative murine mitochondrial transporter homology of hMRS3/4.
- Author
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Li FY, Leibiger B, Leibiger I, and Larsson C
- Subjects
- Amino Acid Sequence genetics, Animals, Blotting, Northern methods, Cation Transport Proteins, Chromosome Mapping methods, DNA, Complementary analysis, Fluorescence, Humans, Mice, Molecular Sequence Data, Transcription, Genetic genetics, Carrier Proteins genetics, Membrane Transport Proteins genetics, Mitochondria genetics, Sequence Homology
- Abstract
We report here a novel murine gene, Mrs3/4, encoding a putative mitochondrial transporter homologous to the yeast mitochondrial RNA splicing protein 3 and 4 (yMRS3&4) and its human counterpart hMRS3/4. By analyses of radiation hybrids, Mrs3/4 was mapped to mouse Chromosome (Chr) 19 between D19Mit66 and D19Mit24 with a distance of 22.4 cR to D19Mit66, a region that is syntenic to human Chromosome 10q24, where the human gene is located. Structural analysis shows that these proteins belong to the mitochondrial carrier family (MCF), which is characterized by three repeats with two transmembrane domains in each repeat. The murine Mrs3/4 gene has two splicing forms similar to the human homolog. The long form corresponds to the 341-amino acid (aa) protein with six transmembrane domains, and the short form corresponds to the 177-aa protein with three transmembrane domains. Both forms have well-conserved signature sequences of MCF. Targeting experiments showed that both forms have mitochondrial localization. Northern blot analyses showed that Mrs3/4 is ubiquitously expressed as a 1.8-kb transcript with a relative abundance in the heart, liver, kidney, and testis. In conclusion, the reported Mrs3/4 gene shows a strong conservation, mitochondrial protein localization, and a ubiquitous expression, which suggests that it has an important role in mammalian cells, most likely as an ion transporter across the mitochondrial inner membrane.
- Published
- 2002
- Full Text
- View/download PDF
40. Characterization of a novel human putative mitochondrial transporter homologous to the yeast mitochondrial RNA splicing proteins 3 and 4.
- Author
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Li FY, Nikali K, Gregan J, Leibiger I, Leibiger B, Schweyen R, Larsson C, and Suomalainen A
- Subjects
- Amino Acid Sequence, Base Sequence, Carrier Proteins metabolism, Chromosome Mapping, DNA, Complementary, Fungal Proteins genetics, Gene Expression Profiling, Humans, Mitochondrial Proteins, Molecular Sequence Data, Mutagenesis, Ophthalmoplegia, Chronic Progressive External genetics, RNA Splicing, Saccharomyces cerevisiae genetics, Spinocerebellar Ataxias genetics, Tissue Distribution, Transfection, Alternative Splicing, Carrier Proteins genetics, Cation Transport Proteins, Chromosomes, Human, Pair 10, Membrane Transport Proteins, Mitochondria metabolism, Repressor Proteins, Saccharomyces cerevisiae Proteins
- Abstract
We report here a novel human gene, hMRS3/4, encoding a putative mitochondrial transporter structurally and functionally homologous to the yeast mitochondrial RNA splicing proteins 3 and 4. These proteins belong to the family of mitochondrial carrier proteins (MCF) and are likely to function as solute carriers. hMRS3/4 spans approximately 10 kb of genomic DNA on chromosome 10q24 and consists of four exons that encode a 364-aa protein with six transmembrane domains. A putative splice variant, encoding a 177-aa protein with three transmembrane domains, was also identified. hMRS3/4 has a well-conserved signature sequence of MCF and is targeted into the mitochondria. When expressed in yeast, hMRS3/4 efficiently restores the mitochondrial functions in mrs3(o)mrs4(o) knock-out mutants. Ubiquitous expression in human tissues and a well-conserved structure and function suggest an important role for hMRS3/4 in human cells.
- Published
- 2001
- Full Text
- View/download PDF
41. Selective insulin signaling through A and B insulin receptors regulates transcription of insulin and glucokinase genes in pancreatic beta cells.
- Author
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Leibiger B, Leibiger IB, Moede T, Kemper S, Kulkarni RN, Kahn CR, de Vargas LM, and Berggren PO
- Subjects
- Calcium Channel Blockers pharmacology, Calcium Channels, L-Type metabolism, Cell Line, Cells, Cultured, Glucose metabolism, Glucose pharmacology, Islets of Langerhans enzymology, Islets of Langerhans metabolism, Models, Biological, Phosphatidylinositol 3-Kinases metabolism, Phosphoinositide-3 Kinase Inhibitors, Promoter Regions, Genetic genetics, Proto-Oncogene Proteins antagonists & inhibitors, Proto-Oncogene Proteins metabolism, Proto-Oncogene Proteins c-akt, RNA Stability, RNA, Messenger genetics, RNA, Messenger metabolism, Ribosomal Protein S6 Kinases antagonists & inhibitors, Ribosomal Protein S6 Kinases metabolism, Substrate Specificity, Glucokinase genetics, Insulin genetics, Insulin pharmacology, Islets of Langerhans drug effects, Protein Serine-Threonine Kinases, Receptor, Insulin metabolism, Signal Transduction drug effects, Transcription, Genetic drug effects
- Abstract
Insulin signaling is mediated by a complex network of diverging and converging pathways, with alternative proteins and isoforms at almost every step in the process. We show here that insulin activates the transcription of its own gene and that of the beta cell glucokinase gene (betaGK) by different mechanisms. Whereas insulin gene transcription is promoted by signaling through insulin receptor A type (Ex11-), PI3K class Ia, and p70s6k, insulin stimulates the betaGK gene by signaling via insulin receptor B type (Ex11+), PI3K class II-like activity, and PKB (c-Akt). Our data provide evidence for selectivity in insulin action via the two isoforms of the insulin receptor, the molecular basis being preferential signaling through different PI3K and protein kinases.
- Published
- 2001
- Full Text
- View/download PDF
42. Glucose-stimulated insulin biosynthesis depends on insulin-stimulated insulin gene transcription.
- Author
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Leibiger B, Wahlander K, Berggren PO, and Leibiger IB
- Subjects
- Animals, Gene Expression Regulation, Insulin Secretion, Models, Genetic, Protein Processing, Post-Translational, RNA Processing, Post-Transcriptional, Rats, Rats, Wistar, Transcription, Genetic, Up-Regulation, Glucose pharmacology, Insulin metabolism, Islets of Langerhans drug effects, Proinsulin biosynthesis
- Abstract
Glucose stimulation of pancreatic beta-cells leads to insulin secretion as well as up-regulation of insulin biosynthesis. The acute elevation in pro-insulin levels is thought to be exclusively because of the activation of translation of pre-existing prepro-insulin mRNA. Glucose-stimulated insulin gene transcription is believed to be a long term effect and should therefore not contribute to the acute elevation in pro-insulin levels. We have recently shown that glucose activates insulin gene transcription within minutes and that secreted insulin is one of the key factors triggering this process in an autocrine manner. We now provide evidence that 50% of the glucose-stimulated, acute pro-insulin biosynthesis within 30 min results from up-regulated insulin gene transcription. Our data led us to propose that glucose elevates pro-insulin levels by stimulating both transcriptional and post-transcriptional/post-translational events to an equal extent. Whereas the stimulatory effect on transcription is mediated by insulin secreted in response to glucose, glucose directly stimulates the post-transcriptional/post-translational processes.
- Published
- 2000
- Full Text
- View/download PDF
43. Identification of a nuclear localization signal, RRMKWKK, in the homeodomain transcription factor PDX-1.
- Author
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Moede T, Leibiger B, Pour HG, Berggren P, and Leibiger IB
- Subjects
- Amino Acid Sequence, Animals, Biological Transport, Cell Nucleus metabolism, DNA, Complementary genetics, Dexamethasone pharmacology, Humans, Mice, Microscopy, Confocal, Molecular Sequence Data, Mutagenesis, Site-Directed, Phosphorylation, Protein Processing, Post-Translational, Protein Structure, Tertiary, Rats, Receptors, Glucocorticoid metabolism, Trans-Activators genetics, Trans-Activators metabolism, Homeodomain Proteins, Trans-Activators chemistry
- Abstract
Pancreatic duodenal homeobox-containing transcription factor 1 (PDX-1) plays a crucial role in pancreas development and beta-cell gene regulation. Absence of PDX-1 leads to pancreas agenesis and its malfunction causes MODY4 diabetes mellitus. PDX-1 has been suggested to be involved in the glucose-dependent regulation of insulin gene transcription. Whereas DNA-binding and transactivation domains of PDX-1 are in the process of being characterized, protein sequences responsible for its nuclear translocation remain unknown. By combining site-directed mutagenesis of putative phosphorylation sites and nuclear localization signal (NLS) motifs with on-line monitoring of GFP-tagged PDX-1 translocation, we demonstrate that the NLS motif RRMKWKK is necessary and in conjunction with the integrity of the 'helix 3' domain of the PDX-1 homeodomain is sufficient for the nuclear import of PDX-1. Furthermore, we show that there is no glucose-dependent cytoplasmic-nuclear cycling of PDX-1.
- Published
- 1999
- Full Text
- View/download PDF
44. Syntaxin 1 interacts with the L(D) subtype of voltage-gated Ca(2+) channels in pancreatic beta cells.
- Author
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Yang SN, Larsson O, Bränström R, Bertorello AM, Leibiger B, Leibiger IB, Moede T, Köhler M, Meister B, and Berggren PO
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, Antigens, Surface immunology, Antigens, Surface isolation & purification, Calcium metabolism, Calcium Channels genetics, Calcium Channels isolation & purification, Calcium Channels, L-Type, Cell Membrane physiology, Cell Membrane ultrastructure, Centrifugation, Density Gradient, Exocytosis physiology, Green Fluorescent Proteins, Luminescent Proteins genetics, Macromolecular Substances, Mice, Mice, Obese, Nerve Tissue Proteins immunology, Nerve Tissue Proteins isolation & purification, Patch-Clamp Techniques, Recombinant Fusion Proteins metabolism, Syntaxin 1, Tubulin immunology, Tubulin physiology, Antigens, Surface metabolism, Calcium Channels physiology, Islets of Langerhans physiology, Nerve Tissue Proteins metabolism
- Abstract
Interaction of syntaxin 1 with the alpha(1D) subunit of the voltage-gated L type Ca(2+) channel was investigated in the pancreatic beta cell. Coexpression of the enhanced green fluorescent protein-linked alpha(1D) subunit with the enhanced blue fluorescent protein-linked syntaxin 1 and Western blot analysis together with subcellular fractionation demonstrated that the alpha(1D) subunit and syntaxin 1 were colocalized in the plasma membrane. Furthermore, the alpha(1D) subunit was coimmunoprecipitated efficiently by a polyclonal antibody against syntaxin 1. Syntaxin 1 also played a central role in the modulation of L type Ca(2+) channel activity because there was a faster Ca(2+) current run-down in cells incubated with antisyntaxin 1 compared with controls. In parallel, antisyntaxin 1 markedly reduced insulin release in both intact and permeabilized cells, subsequent to depolarization with K(+) or exposure to high Ca(2+). Exchanging Ca(2+) for Ba(2+) abolished the effect of antisyntaxin 1 on both Ca(2+) channel activity and insulin exocytosis. Moreover, antisyntaxin 1 had no significant effects on Ca(2+)-independent insulin release trigged by hypertonic stimulation. This suggests that there is a structure-function relationship between the alpha(1D) subunit of the L type Ca(2+) channel and the exocytotic machinery in the pancreatic beta cell.
- Published
- 1999
- Full Text
- View/download PDF
45. Cysteine string protein (CSP) is an insulin secretory granule-associated protein regulating beta-cell exocytosis.
- Author
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Brown H, Larsson O, Bränström R, Yang SN, Leibiger B, Leibiger I, Fried G, Moede T, Deeney JT, Brown GR, Jacobsson G, Rhodes CJ, Braun JE, Scheller RH, Corkey BE, Berggren PO, and Meister B
- Subjects
- Animals, Calcium Channels metabolism, Cell Fractionation, Cells, Cultured, Cytoplasmic Granules ultrastructure, Fluorescent Antibody Technique, HSP40 Heat-Shock Proteins, Insulin Secretion, Islets of Langerhans ultrastructure, Male, Membrane Proteins genetics, Membrane Proteins immunology, Mice, Patch-Clamp Techniques, RNA, Messenger isolation & purification, Rats, Rats, Sprague-Dawley, Recombinant Proteins biosynthesis, Recombinant Proteins immunology, Subcellular Fractions chemistry, Cytoplasmic Granules chemistry, Exocytosis, Insulin metabolism, Islets of Langerhans metabolism, Membrane Proteins isolation & purification
- Abstract
Cysteine string proteins (CSPs) are novel synaptic vesicle-associated protein components characterized by an N-terminal J-domain and a central palmitoylated string of cysteine residues. The cellular localization and functional role of CSP was studied in pancreatic endocrine cells. In situ hybridization and RT-PCR analysis demonstrated CSP mRNA expression in insulin-producing cells. CSP1 mRNA was present in pancreatic islets; both CSP1 and CSP2 mRNAs were seen in insulin-secreting cell lines. Punctate CSP-like immunoreactivity (CSP-LI) was demonstrated in most islets of Langerhans cells, acinar cells and nerve fibers of the rat pancreas. Ultrastructural analysis showed CSP-LI in close association with membranes of secretory granules of cells in the endo- and exocrine pancreas. Subcellular fractionation of insulinoma cells showed CSP1 (34/36 kDa) in granular fractions; the membrane and cytosol fractions contained predominantly CSP2 (27 kDa). The fractions also contained proteins of 72 and 70 kDa, presumably CSP dimers. CSP1 overexpression in INS-1 cells or intracellular administration of CSP antibodies into mouse ob/ob beta-cells did not affect voltage-dependent Ca2+-channel activity. Amperometric measurements showed a significant decrease in insulin exocytosis in individual INS-1 cells after CSP1 overexpression. We conclude that CSP is associated with insulin secretory granules and that CSP participates in the molecular regulation of insulin exocytosis by mechanisms not involving changes in the activity of voltage-gated Ca2+-channels.
- Published
- 1998
- Full Text
- View/download PDF
46. Short-term regulation of insulin gene transcription by glucose.
- Author
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Leibiger B, Moede T, Schwarz T, Brown GR, Köhler M, Leibiger IB, and Berggren PO
- Subjects
- Animals, Base Sequence, Cricetinae, DNA Primers, Green Fluorescent Proteins, Luminescent Proteins genetics, Proinsulin genetics, Protein Precursors genetics, RNA, Messenger genetics, Recombinant Fusion Proteins genetics, Tumor Cells, Cultured, Gene Expression Regulation drug effects, Glucose pharmacology, Insulin pharmacology, Transcription, Genetic drug effects
- Abstract
Whereas short-term regulation of insulin biosynthesis at the level of translation is well accepted, glucose-dependent transcriptional control is still believed to be a long-term effect occurring after more than 2 hr of glucose stimulation. Because pancreatic beta cells are exposed to elevated glucose levels for minutes rather than hours after food uptake, we hypothesized the existence of a short-term transcriptional control. By studying the dynamics of newly synthesized (prepro)insulin RNA and by employing on-line monitoring of gene expression in single, insulin-producing cells, we were able to provide convincing evidence that insulin gene transcription indeed is affected by glucose within minutes. Exposure of insulinoma cells and isolated pancreatic islets to elevated glucose for only 15 min resulted in a 2- to 5-fold elevation in (prepro)insulin mRNA levels within 60-90 min. Similarly, insulin promoter-driven green fluorescent protein expression in single insulin-producing cells was significantly enhanced after transient glucose stimulation. Thus, short-term signaling, such as that involved in insulin secretion, also may regulate insulin gene transcription.
- Published
- 1998
- Full Text
- View/download PDF
47. In situ activation of the type 2 ryanodine receptor in pancreatic beta cells requires cAMP-dependent phosphorylation.
- Author
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Islam MS, Leibiger I, Leibiger B, Rossi D, Sorrentino V, Ekström TJ, Westerblad H, Andrade FH, and Berggren PO
- Subjects
- Animals, Biological Transport, Mice, Mice, Obese, Phosphorylation, Caffeine pharmacology, Calcium metabolism, Central Nervous System Stimulants pharmacology, Cyclic AMP metabolism, Islets of Langerhans metabolism, Ryanodine Receptor Calcium Release Channel metabolism, Signal Transduction drug effects
- Abstract
Molecular mechanisms that regulate in situ activation of ryanodine receptors (RY) in different cells are poorly understood. Here we demonstrate that caffeine (10 mM) released Ca2+ from the endoplasmic reticulum (ER) in the form of small spikes in only 14% of cultured fura-2 loaded beta cells from ob/ob mice. Surprisingly, when forskolin, an activator of adenylyl cyclase was present, caffeine induced larger Ca2+ spikes in as many as 60% of the cells. Forskolin or the phosphodiesterase-resistant PKA activator Sp-cAMPS alone did not release Ca2+ from ER. 4-Chloro-3-ethylphenol (4-CEP), an agent that activates RYs in other cell systems, released Ca2+ from ER, giving rise to a slow and small increase in [Ca2+]i in beta cells. Prior exposure of cells to forskolin or caffeine (5 mM) qualitatively altered Ca2+ release by 4-CEP, giving rise to Ca2+ spikes. In glucose-stimulated beta cells forskolin induced Ca2+ spikes that were enhanced by 3,9-dimethylxanthine, an activator of RYs. Analysis of RNA from islets and insulin-secreting betaTC-3-cells by RNase protection assay, using type-specific RY probes, revealed low-level expression of mRNA for the type 2 isoform of the receptor (RY2). We conclude that in situ activation of RY2 in beta cells requires cAMP-dependent phosphorylation, a process that recruits the receptor in a functionally operative form.
- Published
- 1998
- Full Text
- View/download PDF
48. Exocytosis of insulin promotes insulin gene transcription via the insulin receptor/PI-3 kinase/p70 s6 kinase and CaM kinase pathways.
- Author
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Leibiger IB, Leibiger B, Moede T, and Berggren PO
- Subjects
- Animals, Calcium-Calmodulin-Dependent Protein Kinase Type 2, Cells, Cultured, Exocytosis genetics, Gene Expression Regulation, Enzymologic drug effects, Gene Expression Regulation, Enzymologic physiology, Glucose pharmacology, Insulin metabolism, Insulin pharmacology, Islets of Langerhans chemistry, Islets of Langerhans cytology, Islets of Langerhans metabolism, Phosphatidylinositol 3-Kinases metabolism, Rats, Receptor, Insulin metabolism, Ribosomal Protein S6 Kinases metabolism, Signal Transduction genetics, Transcription, Genetic physiology, Transfection, Calcium-Calmodulin-Dependent Protein Kinases metabolism, Insulin genetics, Phosphatidylinositol 3-Kinases genetics, Receptor, Insulin genetics, Ribosomal Protein S6 Kinases genetics
- Abstract
The control of glucose homeostasis by insulin requires, in addition to the glucose-induced insulin release, a highly dynamic control of insulin biosynthesis. Although elevated glucose concentrations have been shown to trigger insulin biosynthesis at the levels of transcription and translation, the molecular mechanisms underlying the immediate transcriptional control are poorly understood. By investigating signal transduction pathways involved in the "glucose-dependent" transcriptional control, thereby analyzing endogenous (prepro)insulin mRNA levels and monitoring on-line insulin promoter-driven GFP expression, we provide, for the first time, evidence that physiologically stimulated insulin secretion from the pancreatic beta cell promotes insulin biosynthesis by enhancing insulin gene transcription in an autocrine manner. We show that secreted insulin acts via beta-cell insulin receptors and up-regulates insulin gene transcription by signaling through the IRS-2/PI-3 kinase/p70 s6k and CaM kinase pathways.
- Published
- 1998
- Full Text
- View/download PDF
49. Mapping of the human Ca2+ channel beta 4 subunit to 2q22-23 and its expression in developing mouse.
- Author
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Betz R, Leibiger B, Farnebo F, Lagercrantz S, Piehl F, Leibiger I, and Larsson C
- Subjects
- Animals, Chromosome Mapping, Cricetinae, Humans, Hybrid Cells, In Situ Hybridization, Mice, Polymerase Chain Reaction, Calcium Channels genetics, Chromosomes, Human, Pair 2, Gene Expression Regulation, Developmental
- Published
- 1998
- Full Text
- View/download PDF
50. A silencer and an adjacent positive element interact to modulate the activity of the human insulin promoter.
- Author
-
Clark AR, Wilson ME, Leibiger I, Scott V, and Docherty K
- Subjects
- Animals, Base Sequence, Binding Sites, Cell Line, Cricetinae, DNA genetics, DNA metabolism, Homeodomain Proteins metabolism, Host Cell Factor C1, Humans, Islets of Langerhans metabolism, Molecular Sequence Data, Mutagenesis, Site-Directed, Octamer Transcription Factor-1, Plasmids genetics, Protein Binding, Transcription Factors metabolism, DNA-Binding Proteins, Genes, Regulator, Insulin genetics, Promoter Regions, Genetic
- Abstract
A negative regulatory element (NRE) is located between positions -279 and -261 relative to the transcription start site of the human insulin gene. The NRE contains at least three distinct overlapping binding sites for several nuclear proteins. These proteins may be distinguished by their interaction with mutant variants of the NRE. Mutagenesis of two of these protein-binding sites within an insulin gene fragment containing sequences from position -361 to position +112 attenuated the negative activity of the NRE, confirming the importance of these sites in the function of the NRE. When placed in isolation upstream of the herpes-simplex-virus thymidine-kinase promoter, the NRE exhibited stimulatory activity in non-beta (BHK) and beta (HIT) cells. The positive activity within the NRE was mapped to a sequence that resembled the binding site for the ubiquitous factor Oct-1. These results indicated that the negative activity of the NRE was dependent on its interaction with other regulatory sites within the insulin gene. Thus, the NRE exhibited negative activity in transfected HIT cells when placed upstream of an insulin gene fragment (positions -261 to +112). However, its activity was modulated in a positive manner by inclusion of additional sequences from position -279 to -341. This region contains the CT3 box that binds the homeodomain protein IUF1 (insulin upstream factor 1). The NRE resembled a silencer, being at least partly independent of precise location and orientation, and being able to operate upon a variety of promoters. The role of the NRE is unclear, although it may be involved in restricting expression of the insulin gene to cells of the islets of Langerhans.
- Published
- 1995
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