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2. F4-related mutation and expression analysis of the aminopeptidase N gene in pigs1

Author

Dieter Deforce, Luc Peelman, Vesna Melkebeek, Van Ut Nguyen, M. Van Poucke, Eric Cox, Tiphanie Goetstouwers, and Annelies Coddens

Subjects
pig, Untranslated region, F4 receptor, Genotype, Swine, Fimbria, CD13 Antigens, Biology, medicine.disease_cause, Bacterial Adhesion, Gene Expression Regulation, Enzymologic, Molecular Genetics, Enterotoxigenic Escherichia coli, Genetics, medicine, TaqMan, Animals, Genetic Predisposition to Disease, Receptor, Escherichia coli Infections, Mutation, General Medicine, aminopeptidase N, F4 phenotyping, Molecular biology, Reverse transcriptase, Small intestine, medicine.anatomical_structure, F4 enterotoxigenic Escherichia coli, Animal Science and Zoology, Animal Genetics, Food Science
Abstract

Intestinal infections with F4 enterotoxigenic Escherichia coli (ETEC) are worldwide an important cause of diarrhea in neonatal and recently weaned pigs. Adherence of F4 ETEC to the small intestine by binding to specific receptors is mediated by F4 fimbriae. Porcine aminopeptidase N (ANPEP) was recently identified as a new F4 receptor. In this study, 7 coding mutations and 1 mutation in the 3′ untranslated region (3' UTR)were identified in ANPEP by reverse transcriptase (RT–) PCR and sequencing using 3 F4 receptor-positive (F4R+) and 2 F4 receptor-negative (F4R–) pigs, which were F4 phenotyped based on the MUC4 TaqMan, oral immunization, and the in vitro villous adhesion assay. Three potential differential mutations (g.2615C > T, g.8214A > G, and g.16875C > G) identified by comparative analysis between the 3 F4R+ and 2 F4R– pigs were genotyped in 41 additional F4 phenotyped pigs. However, none of these 3 mutations could be associated with F4 ETEC susceptibility. In addition, the RT-PCR experiments did not reveal any differential expression or alternative splicing in the small intestine of F4R+ and F4R– pigs. In conclusion, we hypothesize that the difference in F4 binding to ANPEP is due to modifications in its carbohydrate moieties.

Published
2014
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3. SNP identification, linkage and radiation hybrid mapping of the porcine lamin A/C (LMNA) gene to chromosome 4q

Author

Antonín Stratil, D. Wagenknecht, Luc Peelman, H. Bartenschlager, Hermann Geldermann, I. Majzlík, and M. Van Poucke

Subjects
Genetics, Radiation Hybrid Mapping, congenital, hereditary, and neonatal diseases and abnormalities, integumentary system, Genetic Linkage, Swine, Single-nucleotide polymorphism, General Medicine, Quantitative trait locus, Biology, Lamin Type A, Chromosomes, Mammalian, Polymorphism, Single Nucleotide, Molecular biology, LMNA, Restriction site, Chromosome 4, Food Animals, Genetic linkage, Animals, Animal Science and Zoology, Radiation hybrid mapping, Allele frequency, Alleles
Abstract

Summary The lamins are components of nuclear lamina and they have a profound influence on nuclear structure and functions. They are encoded by three genes, LMNA, LMNB1 and LMNB2. A genomic fragment of the porcine LMNA gene (822 bp; from exons 7 to 9) was amplified by polymerase chain reaction and comparatively sequenced. Four single nucleotide polymorphisms (SNPs) were identified in intronic sequences: G162A, G208A, T367G and C618T. The SNPs are within the restriction sites for enzymes Bsh1236I, HpaII, AluI and Bsh1236I respectively. Allele frequencies at SNPs G208A, T367G and C618T were determined by using eight pig breeds. Linkage analysis in the Hohenheim Meishan × Pietrain family placed the LMNA gene in the chromosome 4q linkage group, between MEF2D and GBA (MEF2D– 3.0 cM –LMNA– 0.2 cM –GBA). In radiation hybrid mapping LMNA was most significantly linked to SW270 on chromosome 4 (39 cR; LOD = 7.86). The LMNA gene is located in the quantitative trait loci region for some carcass traits on chromosome 4q.

Published
2006
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4. Characterization of the porcine melanocortin 2 receptor gene (MC2R )

Author

Patrick Chardon, Gary A. Rohrer, M. Van Poucke, Marc Mattheeuws, K. Jacobs, A. Van Zeveren, Martine Yerle, and Luc Peelman

Subjects
Genetics, Bacterial artificial chromosome, General Medicine, Biology, Molecular biology, Loss of heterozygosity, Exon, Coding region, Animal Science and Zoology, Melanocortin, Allele frequency, Gene, Melanocortin 5 receptor
Abstract

A porcine bacterial artificial chromosome (BAC) clone, containing the melanocortin 2 receptor gene (MC2R) was isolated. The complete coding sequence of the MC2R gene, contained in 1 exon, was determined. Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) was performed on a 241-bp coding fragment. An AluI polymorphism, detecting a silent mutation, was found and typed on unrelated animals of five different pig breeds. The Meishan, Pietrain and Large White breeds differ significantly in allele frequencies from the Landrace and Czech Meat Pig breeds. The melanocortin 5 receptor gene (MC5R) was detected by PCR in the same BAC clone, as could be expected from the human and porcine mapping data. PCR-SSCP was performed on a 200-bp coding of MC5R, but no polymorphisms were detected. The BAC clone was mapped to Sscr6q27 by fluorescent in situ hybridization (FISH). A (CA)n microsatellite (SGU0002), isolated from the BAC, was localized on chromosome 6 by RH mapping near marker SW1473 and by linkage mapping on the MARC reference family at the same position as the marker SW2173 (97 cM). Allele frequencies, heterozygosity and polymorphism information contents (PIC) values were calculated for the five different pig breeds examined. The transcription of both genes in porcine liver, heart, kidney, fat, brain, pancreas, stomach, bladder, ovaries, lung, spleen, skin, adrenal gland and muscle tissues was examined by reverse transcriptase-polymerase chain reaction. Transcription was detected in skin and adrenal gland tissues for MC2R, while a positive signal was detected for MC5R in kidney, fat, pancreas, skin, adrenal gland and spleen tissues.

Published
2002
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5. Characterization of the porcine FABGL gene

Author

A. Van Zeveren, K. Jacobs, Luc Peelman, M. Van Poucke, and Marc Mattheeuws

Subjects
Genetics, Candidate gene, biology, General Medicine, Reductase, Major histocompatibility complex, medicine.disease_cause, Molecular biology, Restriction site, Transcription (biology), biology.protein, medicine, Animal Science and Zoology, Gene, Escherichia coli, Peptide sequence
Abstract

The porcine major histocompatibility complex, also called swine lymphocyte antigen (SLA) complex, is of particular interest not only because of its central role in the immune response, but also because of its influence on many traits such as reproduction, fatness and meat quality. The porcine FABGL (FabG (beta-ketoacyl-[acyl-carrierprotein] reductase, Escherichia coli) like) gene, coding for a 17beta-hydroxysteroid dehydrogenase (17beta-HSD), is a candidate gene for these traits. The complete gene was sequenced and compared with human and mouse FABGL sequences. The deduced amino acid sequence showed 85 and 83% sequence identity to human and mouse sequences, respectively. Polymorphicic BbvI and DdeI restriction sites were found in the porcine FABGL gene. The promoter was compared with the promoter regions of human and mouse FABGL sequence in order to identify putative regulatory elements. The transcription profile of the porcine gene was determined and showed a widespread tissue distribution.

Published
2002
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6. Contents Vol. 102, 2003

Author

H.A. Lewin, G. Pielberg, H.B. Park, V. Russo, R.W. Dunstan, M. Niikura, A. Fu, H. Hayes, C. Moran, J. Womack, S. Kubickova, M.A. Ferguson-Smith, J.L. Williams, J.L. Myka, J. van der Poel, Loren C. Skow, B. van Hest, M. Menotti-Raymond, David L. Adelson, J. Schläpfer, H. Bird, J.R. Garbe, A.A. Schäffer, N. Yasinetskaya, M. Marra, K. Jacobs, C. Elduque, Y. Da, F. Yang, E.G. Cothran, W.J. Murphy, D. Zudova, P. Chardon, V. Amarger, E.P. Cribiu, Y. Zhang, D. Gallagher, R. Rullo, L.B. Madsen, M. Zanotti, L. Krejci, L. Molteni, M. Van Poucke, L. Harman, J.W. Keele, M.M. Binns, C. Genêt, J.N. Derr, J. Schein, B. Backofen, S. Peto, G. Guérin, Donald Miller, F.A. Ponce de León, R. Voß-Nemitz, S. Kiuchi, J.A.M. Graves, S. Cirera, Thomas Faraut, C. Garcia, P.J. de Jong, A.L. Gustafson, O.A. Ryder, W.-S. Liu, R. Stephens, J. Rubes, A. Crisà, S. Braglia, T.L. Lear, C.B. Jørgensen, M.V. Arruga, M.F. Rothschild, H. Uenishi, N.P. Carter, O. Ryder, J.A. Price, N. Iannuccelli, Samodha C. Fernando, C. Baiocco, M. Dunø, K. Benke, L.M. Daniels, J.R. Mickelson, A. Valentini, K.M. Reed, K.E. Murphy, B. Benkel, O.R.P. Bininda-Emonds, G. Brown, H.H. Cheng, D. Milenkovic, B. Koop, B. Brenig, Doris M. Kupfer, S. So, L. Iannuzzi, V. Fillon, Y. Chen, A.T.V. Pillai, M.L. Cox, C.W. Beattie, M.L. Houck, N.E. Raney, C. Drögemüller, N. Bosak, Jillian F. Maddox, L. Fontanesi, T.L. Ward, L.D. Chaves, B.P. Chowdhary, L. Alexander, H.-C. Liu, R. Erlandsson, L. Nanni Costa, C.A. Gill, M. Mattheeuws, T. Bønsdorff, N. Rogalska-Niznik, R. Agarwala, M. Schmid, B. Lama, K.A. Greer, A. Van Zeveren, Y. Meng, G. Rohrer, M. Yerle, J.E. Fulton, M. Breen, L.J. Peelman, C. Li, H. Yasue, B. Fu, H.P. Klinger, R.P.M.A. Crooijmans, V. Cantegrel, E. Bailey, T. Veenendaal, S. Mikawa, V.D. Rilington, S. Marklund, M.C. Savarese, S. Mashima, B.J. Ostroski, I. Szczerbal, H. Fiegler, A.H. Petersen, A. Vignal, D.E. Harry, K. Osoegawa, J. Klukowska, H. Hiraiwa, L. Chemnick, L. Schibler, J. Aldenhoven, A. Jaadar, B.W. Kirkpatrick, C. Hansen, V.A. David, M. Longeri, S.I. Anderson, F. Kasai, E. Scotti, S.S. Moore, G. Bongioni, D. Incarnato, S.J. Valberg, A. Eggen, I.R. Franklin, T. Zharkikh, J. Quackenbush, L.D. Lieto, M. Gautier, C. Marchitelli, Bruce A. Roe, E. Cribiu, K.M. Credille, E.J. Cargill, R. Middleton, M .A.M. Groenen, Y. Palti, O. Rezacova, N. Ramlachan, W. Zimmermann, R. Fries, Terje Raudsepp, J.-T. Jeon, C.W. Ernst, E.A. Ostrander, F. Martins-Wess, Udaya DeSilva, L. Andersson, C.E. Rexroad, A. Winter, C.M. Seabury, Y. Lee, M.N. Romanov, V.H. Nielsen, W. Rens, R.D. Schnabel, M.E. Delany, K. Hemmatian, R. Guyon, S.E. Swanberg, M. van Eckeveld, C. Schelling, B.S.D. Urquhart, A. Galli, P.J. Venta, J.F. Taylor, S. Cornelissen, J.B. Dodgson, K.L. Tsai, K. Sandberg, L. Ferretti, G.P. Di Meo, Rebecca L. Tallmadge, Bhanu P. Chowdhary, W. Bridge, C.R. Farber, C. Penado, Alan Archibald, D. Milan, R. Davoli, T.J. Robinson, H. Fairclough, P.C.M. O’Brien, M. Switonski, F. Galibert, T. Leeb, S.J. O’Brien, T. Hayashi, F.A. Habermann, A. Perucatti, Fares Z. Najar, T. Lear, C. Bendixen, Douglas F. Antczak, B. Thomsen, L. Buttazzoni, J. Aerts, J.E. Swinburne, R. Thomas, X. Guo, S.A. Gahr, M. Fredholm, P.H. Nissen, A. Robic, I. Edfors-Lilja, S. Karamycheva, G. Dolf, L.A. Clark, Y. Lahbib-Mansais, L. Skow, Noelle E. Cockett, G.Q. Fitch, and A.T. Bowling

Subjects
Botany, Genetics, Biology, Molecular Biology, Genetics (clinical)
Published
2003
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7. Subject Index Vol. 102, 2003

Author

P.H. Nissen, I. Edfors-Lilja, J. Aerts, R. Thomas, X. Guo, M.A. Ferguson-Smith, L. Molteni, M. Fredholm, B. Koop, P.J. Venta, H. Bird, J.N. Derr, T. Zharkikh, J. Quackenbush, M. Marra, R.W. Dunstan, H.A. Lewin, G. Pielberg, L. Chemnick, L.A. Clark, Y. Lahbib-Mansais, N. Ramlachan, Noelle E. Cockett, A. Fu, H. Hayes, G.Q. Fitch, H. Fiegler, A.T. Bowling, E.P. Cribiu, D. Gallagher, R. Rullo, B. Backofen, J.-T. Jeon, C.W. Ernst, H.P. Klinger, R.P.M.A. Crooijmans, H. Yasue, J.L. Williams, V.D. Rilington, B. Fu, G. Rohrer, H.B. Park, V.A. David, F. Kasai, D.E. Harry, M. Zanotti, V. Amarger, R. Fries, Terje Raudsepp, V. Russo, D. Incarnato, Loren C. Skow, I.R. Franklin, C. Marchitelli, S. Cirera, S.S. Moore, M.C. Savarese, K. Sandberg, S. Kubickova, J.F. Taylor, S. Cornelissen, L. Krejci, B.J. Ostroski, A.A. Schäffer, Thomas Faraut, K. Osoegawa, J. Klukowska, M. Gautier, C. Elduque, S. Mikawa, S. Marklund, Samodha C. Fernando, G. Guérin, Donald Miller, F.A. Ponce de León, A. Jaadar, F. Martins-Wess, C.E. Rexroad, Udaya DeSilva, L. Ferretti, K.L. Tsai, L.B. Madsen, E. Bailey, J. Schläpfer, E.A. Ostrander, G.P. Di Meo, J. van der Poel, Bruce A. Roe, J.L. Myka, E. Scotti, H.H. Cheng, E.G. Cothran, G. Bongioni, L. Andersson, A. Winter, M.E. Delany, W.J. Murphy, D. Zudova, T.L. Ward, C. Baiocco, A. Crisà, Rebecca L. Tallmadge, W. Rens, J. Aldenhoven, Bhanu P. Chowdhary, K.E. Murphy, M.F. Rothschild, R.D. Schnabel, M.L. Cox, J.R. Mickelson, A. Valentini, M. Niikura, V. Fillon, M.V. Arruga, C.M. Seabury, M. van Eckeveld, C. Schelling, C. Genêt, J.W. Keele, B.W. Kirkpatrick, M. Schmid, E. Cribiu, C.R. Farber, C. Li, N. Rogalska-Niznik, Y. Chen, J.A. Price, C. Drögemüller, O.R.P. Bininda-Emonds, L.D. Chaves, M.L. Houck, N.E. Raney, C. Penado, K.M. Credille, Alan Archibald, B. Lama, S.J. Valberg, O. Rezacova, B.S.D. Urquhart, A. Eggen, M. Van Poucke, J.A.M. Graves, L. Iannuzzi, K. Benke, L.M. Daniels, B.P. Chowdhary, G. Brown, M. Yerle, Y. Da, F. Yang, S. Braglia, B. Brenig, M .A.M. Groenen, D. Milenkovic, I. Szczerbal, M. Mattheeuws, A.H. Petersen, Jillian F. Maddox, B. van Hest, W. Zimmermann, J.R. Garbe, R. Stephens, J. Rubes, Y. Lee, J. Schein, L. Alexander, H.-C. Liu, M.N. Romanov, M.M. Binns, M. Dunø, P.J. de Jong, A.L. Gustafson, S. Kiuchi, S.E. Swanberg, Y. Palti, B. Benkel, A. Van Zeveren, Y. Meng, C.B. Jørgensen, A. Galli, K.M. Reed, H. Uenishi, N.P. Carter, V.H. Nielsen, J.B. Dodgson, A.T.V. Pillai, C.W. Beattie, K. Hemmatian, R. Guyon, J.E. Fulton, N. Bosak, R. Erlandsson, A. Vignal, C. Moran, J. Womack, K. Jacobs, O. Ryder, Doris M. Kupfer, S. So, David L. Adelson, Y. Zhang, L. Harman, R. Voß-Nemitz, C. Garcia, K.A. Greer, L.J. Peelman, V. Cantegrel, T. Veenendaal, M. Longeri, S.I. Anderson, A. Robic, S. Karamycheva, R. Middleton, B. Thomsen, N. Iannuccelli, L. Buttazzoni, J.E. Swinburne, L. Fontanesi, L. Nanni Costa, C.A. Gill, H. Hiraiwa, S.A. Gahr, G. Dolf, S. Peto, L. Skow, T.L. Lear, L.D. Lieto, M. Breen, E.J. Cargill, T. Bønsdorff, R. Agarwala, M. Menotti-Raymond, N. Yasinetskaya, P. Chardon, W.-S. Liu, O.A. Ryder, S. Mashima, L. Schibler, C. Hansen, Douglas F. Antczak, S.J. O’Brien, T. Hayashi, F.A. Habermann, A. Perucatti, Fares Z. Najar, T. Lear, C. Bendixen, W. Bridge, D. Milan, R. Davoli, T.J. Robinson, H. Fairclough, P.C.M. O’Brien, M. Switonski, F. Galibert, and T. Leeb

Subjects
Index (economics), Statistics, Genetics, Subject (documents), Biology, Molecular Biology, Genetics (clinical)
Published
2003
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8. An Alw 26I PCR-RFLP in exon 1 of the porcine SKI oncogene and mapping the gene to the RYR1 (CRC ) linkage group on chromosome 6

Author

Luc Peelman, Antonín Stratil, Gerald Reiner, Paolo Zambonelli, M. Van Poucke, Roberta Davoli, Hermann Geldermann, Stratil A., Reiner G., Peelman L.J., Davoli R., Van Poucke M., Zambonelli P., and Geldermann H.

Subjects
Male, Genetic Linkage, Molecular Sequence Data, Sus scrofa, Biology, Polymerase Chain Reaction, Exon, Gene Frequency, Proto-Oncogene Proteins, Proto-Oncogenes, Genetics, Animals, Cloning, Molecular, pig, PCR-RFLP, SKI, SSC6, Gene, Alleles, DNA Primers, Linkage (software), RYR1, Radiation Hybrid Mapping, Base Sequence, Oncogene, Chromosome Mapping, Chromosome, Ryanodine Receptor Calcium Release Channel, General Medicine, musculoskeletal system, Molecular biology, DNA-Binding Proteins, Female, Animal Science and Zoology, Restriction fragment length polymorphism, tissues, human activities, Polymorphism, Restriction Fragment Length
Abstract

An Alw 26I PCR-RFLP in exon 1 of the porcine SKI oncogene and mapping the gene to the RYR1 (CRC ) linkage group on chromosome 6

Published
2002
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11. Contents Vol. 93, 2001

Author

T. Brueckmann, W. Brenner, M. Steinemann, W. Vogel, S. Schlaubitz, C. Zühlke, M. Lombard, F. Boán, K. Benirschke, S. Naumann, S.W. Bremer, C. Steinlein, S. Steinemann, F. Richard, P.D. Thomsen, M. Yerle, K.D. Zang, Z. Docherty, C. Amid, K. Mrasek, I. Schubert, M. Mende, I. Nanda, T. Paiss, C. Genêt, L.J. Peelman, I. Chudoba, M. Hughes, R.-D. Wegner, U. Claussen, L. Sánchez, B. Seipel, F. Grützner, F.J. García-Cozar, D. Prawitt, B.U. Zabel, J.L. Wright, A. Van Zeveren, K. Stout, V. Kalscheuer, M. Stumm, R.V. Rambau, N. Reissmann, D.S. Gallagher, B. Zabel, A. Ishikawa, C. Messaoudi, A.T. Kumamoto, E.C. Akeson, A. Mujica, A. Dalski, P. Kaiser, T. Liehr, J.G. Scammell, S. Bremer, C. Pfeifer, S. Munsche, M.M. Valdivia, M. Van Poucke, M. Schmid, C.M. Tuck-Muller, H. Starke, F. Domínguez, Y. Matsuda, S. Störkel, C.G. Mathew, F.F.B. Elder, S. Narayanswami, H. Scherthan, J. Decker, E. Schwinger, A. Niveleau, V. Trifonov, H. Mayrhofer, J. Gómez-Márquez, J.P. Lambert, S.-E. Bikar, E. Zend-Ajusch, L.J. Bechtel, T. Haaf, Y.A. Wang, A. Viñas, C. Iglesias, C. Mackie Ogilvie, A. Bahr, T. Nagase, A. Dufke, H.H.Q. Heng, A. Winterpacht, W. Lu, T.J. Robinson, C. Maier, K. Matsubara, A. Heller, A. Kuroiwa, M. Rocchi, B. Dutrillaux, C.J. Ye, N. Nomura, N. Rubtsov, E.R. Schmidt, T. Namikawa, M.T. Davisson, C. Tuggle, K. Gardiner, H. Enders, G. Liu, M. Buceta, H. Hanson, H. Hauser, N. Sampson, H. Neitzel, P.D. Waters, T. Hankeln, H. Tönnies, C. Pendón, J. Bolívar, M.L. Houck, D. Reutzel, M. Leipoldt, A. Cichutek, P. Moens, J.A.M. Graves, P.J. Kirby, B. Maurer, A. Astola, S.A. Krawetz, and F. Piumi

Subjects
Botany, Genetics, Biology, Molecular Biology, Genetics (clinical)
Published
2001
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12. Identification of polymorphisms in the ovine Shadow of prion protein (SPRN) gene and assessment of their effect on promoter activity and susceptibility for classical scrapie

Author

A. Van Zeveren, Luc Peelman, Xavier Saelens, M. Van Poucke, Tim Erkens, Luc Duchateau, Evelyne Lampo, and Bert Schepens

Subjects
Untranslated region, Genetics, Candidate gene, Polymorphism, Genetic, Sheep, Base Sequence, Molecular Sequence Data, Promoter, Scrapie, Nerve Tissue Proteins, General Medicine, Biology, Molecular biology, PRNP, Exon, Mutation, Coding region, Animals, Animal Science and Zoology, Genetic Predisposition to Disease, Promoter Regions, Genetic, Gene
Abstract

Shadow of prion protein (SPRN) is an interesting candidate gene thought to be involved in prion pathogenesis. In humans, an association has already been discovered between mutations in SPRN and the incidence of variant and sporadic Creutzfeldt-Jakob disease. However, in sheep, the effect of mutations in SPRN is largely unknown. Therefore, we analysed the presence of mutations in the entire ovine SPRN gene, their association with scrapie susceptibility and their effect on SPRN promoter activity. In total, 26 mutations were found: seven in the promoter region, four in intron 1, seven in the coding sequence and eight in the 3' untranslated region. The mutations detected in the coding sequence and the promoter region were subsequently analysed in more detail. In the coding sequence, a polymorphism causing a deletion of two alanines was found to be associated with susceptibility for classical scrapie in sheep. Furthermore, a functional analysis of deletion constructs of the ovine SPRN promoter revealed that the region 464 to 230 bp upstream of exon 1 (containing a putative AP-2 and putative Sp1 binding sites) is of functional importance for SPRN transcription. Six mutations in the SPRN promoter were also found to alter the promoter activity in vitro. However, no association between any of these promoter mutations and susceptibility for classical scrapie was found.

Published
2009

13. Porcine PPARGC1A (peroxisome proliferative activated receptor gamma coactivator 1A): coding sequence, genomic organization, polymorphisms and mapping

Author

H. Barthenschlager, Gary A. Rohrer, Martine Yerle, A. Van Zeveren, François Piumi, Luc Peelman, M. Van Poucke, Marc Mattheeuws, and K. Jacobs

Subjects
Swine, Peroxisome Proliferative Activated Receptor Gamma, Biology, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Coactivator, Genetics, Coding region, Animals, Humans, Molecular Biology, Genetics (clinical), In Situ Hybridization, Fluorescence, Genomic organization, DNA Primers, Polymorphism, Genetic, Base Sequence, Chromosome Mapping, Introns, Adipose Tissue, Organ Specificity, Trans-Activators, PPARGC1A, Body Temperature Regulation, Transcription Factors
Abstract

We report here the characterisation of porcine PPARGC1A. Primers based on human PPARGC1A were used to isolate two porcine BAC clones. Porcine coding sequences of PPARGC1A were sequenced together with the splice site regions and the 5′ and 3′ regions. Using direct sequencing nine SNPs were found. Allele frequencies were determined in unrelated animals of five different pig breeds. In the MARC Meishan-White Composite resource population, the polymorphism in exon 9 was significantly associated with leaf fat weight. PPARGC1A has been mapped by FISH to SSC8p21. A (CA)n microsatellite (SGU0001) has been localised near marker SWR1101 on chromosome 8 by RH mapping and at the same position as marker KS195 (32.5 cM) by linkage mapping. The AseI (nt857, Asn/Asn489) polymorphism in exon 8 was used to perform linkage analysis in the Hohenheim pedigrees and located the gene in the same genomic region. Transcription of the gene was detected in adipose, muscle, kidney, liver, brain, heart and adrenal gland tissues, which is in agreement with the function of PPARGC1A in adaptive thermogenesis.

Published
2004

14. A refined comparative map between porcine chromosome 13 and human chromosome 3

Author

M. Van Poucke, Marc Mattheeuws, K. Jacobs, Martine Yerle, Luc Peelman, A. Van Zeveren, C Genêt, Patrick Chardon, Laboratoire de radiobiologie et d'étude du génome (LREG), and Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
Chromosomes, Artificial, Bacterial, Genetic Linkage, Swine, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Biology, Polymerase Chain Reaction, Chromosomes, 03 medical and health sciences, Gene mapping, Gene Order, Genetics, Animals, Humans, COMPARATIVE GENETICS, Genomic library, PIGS, Molecular Biology, In Situ Hybridization, Fluorescence, Metaphase, Genetics (clinical), Gene Library, 030304 developmental biology, Synteny, Chromosome 13, Radiation Hybrid Mapping, 0303 health sciences, Bacterial artificial chromosome, 030305 genetics & heredity, MEN, Physical Chromosome Mapping, Molecular biology, PIG CHROMOSOME 13, Chromosome 3, Cytogenetic Analysis, Microsatellite, Human genome, Chromosomes, Human, Pair 3, CHROMOSOME PORCIN 13, Microsatellite Repeats
Abstract

We report here the localisation of BAIAP1 (13q24), HTR1F (13q45), PTPRG (13q23) and UBE1C (13q24) by fluorescence in situ hybridisation (FISH), and BAIAP1 (Swr2114; 21 cR; LOD = 11.03), GATA2 (Sw2448; 37 cR; LOD = 8.26), IL5RA (Swr2114; 64 cR; LOD = 3.85), LMCD1 (Sw2450; 61 cR; LOD = 4.73), MME (CP; 50 cR; LOD = 7.75), RYK (Swc22; 12 cR; LOD = 18.62) and SGU003 (Sw1876; 6 cR; LOD = 16.99) by radiation hybrid (RH) mapping to porcine chromosome 13 (SSC13). The mapping of these 10 different loci (all mapped to human chromosome 3; HSA3) not only confirms the extended conservation of synteny between HSA3 and SSC13, but also defines more precisely the regions with conserved linkage. The syntenic region of the centromeric part of SSC13 was determined by isolating porcine bacterial artificial chromosome (BAC) clones (842D4 and 1031H1) using primers amplifying porcine microsatellite markers S0219 and S0076 (mapped to this region). Sequence comparison of the BAC end sequences with the human genome sequence showed that the centromeric part of SSC13 is homologous with HSA3p24.

Published
2003

15. Integration of porcine chromosome 13 maps

Author

Christopher K. Tuggle, M. Van Poucke, Luc Peelman, A. Van Zeveren, Martine Yerle, François Piumi, C Genêt, Laboratoire de Génétique Cellulaire (LGC), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Laboratoire de radiobiologie et d'étude du génome (LREG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Recherche Agronomique (INRA), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT), and Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
Chromosomes, Artificial, Bacterial, Swine, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Biology, Polymerase Chain Reaction, 03 medical and health sciences, Gene mapping, CHROMOSOMES, Genetics, Animals, Humans, PIGS, Cloning, Molecular, Molecular Biology, Gene, In Situ Hybridization, Fluorescence, Genetics (clinical), 030304 developmental biology, Chromosome 13, Radiation Hybrid Mapping, 0303 health sciences, 030302 biochemistry & molecular biology, Physical Chromosome Mapping, Genes, Chromosome 3, GENETIC MAPS, In situ hybridisation, %22">Fish , Lod Score, human activities
Abstract

In order to expand the comparative map between human chromosome 3 (HSA3) and porcine chromosome 13 (SSC13), seven genes from HSA3 were mapped on SSC13 by fluorescence in situ hybridisation (FISH), viz. ACAA1, ACPP, B4GALT4, LTF, MYLK, PDHB and RARB. With a view to integrating this expanded comparative map with the existing SSC13 linkage map, we used the INRA-University of Minnesota porcine Radiation Hybrid panel (IMpRH) to localize more precisely and to order 15 genes on the SSC13 map, viz. ACPP, ADCY5, APOD, BCHE, CD86, DRD3, GAP43, PCCB, RAF1, RHO, SI, TF, TFRC, TOP2B and ZNF148. In this way, we were able to create an integrated map, containing 38 type I and 81 type II markers, by correlating the linkage, radiation hybrid (RH) and cytogenetic maps of SSC13. This integrated map will give us the opportunity to take maximal advantage of the comparative mapping strategy for positional candidate cloning of genes responsible for economically important traits.

Published
2001

16. Comparative mapping between human chromosome 3 and porcine chromosome 13

Author

Luc Peelman, A. Van Zeveren, A. Törnsten, Bhanu P. Chowdhary, M. Van Poucke, and Marc Mattheeuws

Subjects
Genetic Markers, Rhodopsin, Swine, Molecular Sequence Data, Receptors, Cell Surface, Biology, Chromosomes, Gene mapping, Chromosome regions, Alpha-Globulins, Genetics, Animals, Humans, Molecular Biology, beta-D-Galactoside alpha 2-6-Sialyltransferase, Genetics (clinical), In Situ Hybridization, Fluorescence, Synteny, Chromosome 13, Chromosome, Chromosome Mapping, Sialyltransferases, Rats, DNA-Binding Proteins, Isoenzymes, Subcloning, Chromosome 3, Human genome, Chromosomes, Human, Pair 3, Collagen, Trypsin Inhibitors, Receptors, Calcium-Sensing, Adenylyl Cyclases
Abstract

Zoo-FISH and somatic cell hybrid panels have earlier demonstrated extended synteny conservation between human chromosome 3 (HSA3) and pig chromosome 13 (SSC13). In the present study, eight human genes viz., ADCY5, CASR, COL7A1, COL8A1, ITIH1, RHO, SIAT1 and XPC, spread along the length of HSA3, were chosen for expanding the comparative map between the two chromosomes. Using human and rat cDNAs, or human- and porcine-specific PCR products as probes, 8 porcine lambda clones were isolated. After subcloning and partial sequence determination, identity of the clones with regards to the specific genes was established. The eight type 1 markers thus obtained were biotin labeled and FISH mapped to pig metaphase spreads. All lambda clones localized to SSC13. In combination with the hitherto published mapping data of coding sequences on SSC13, a preliminary comparative status depicting the relative organization of this chromosome with respect to HSA3 was developed. The comparative map thus obtained bears significance in searching for candidate genes of economically important traits mapped to SSC13.

Published
1999

18. Subject Index Vol. 93, 2001

Author

N. Sampson, W. Lu, H. Neitzel, P.D. Waters, T. Brueckmann, W. Brenner, C. Mackie Ogilvie, A. Heller, H. Tönnies, B. Dutrillaux, M. Steinemann, C. Pendón, J. Bolívar, R.-D. Wegner, M. Lombard, C.J. Ye, A. Ishikawa, E. Zend-Ajusch, T. Nagase, Y.A. Wang, F. Richard, A. Kuroiwa, N. Rubtsov, E.R. Schmidt, C. Tuggle, C. Genêt, K. Gardiner, H. Enders, B. Zabel, A. Van Zeveren, M. Rocchi, H. Scherthan, A. Dalski, R.V. Rambau, J.P. Lambert, K. Stout, A. Dufke, B. Seipel, F.F.B. Elder, J.L. Wright, T. Namikawa, H. Hauser, E. Schwinger, M. Buceta, C. Pfeifer, S. Narayanswami, A.T. Kumamoto, H. Starke, V. Kalscheuer, J.G. Scammell, M. Stumm, K. Matsubara, V. Trifonov, C. Amid, K. Mrasek, G. Liu, A. Bahr, T. Haaf, A. Viñas, C. Iglesias, Z. Docherty, H. Mayrhofer, P. Kaiser, S.-E. Bikar, I. Chudoba, C.M. Tuck-Muller, J. Decker, L.J. Bechtel, H.H.Q. Heng, A. Winterpacht, M.M. Valdivia, C. Messaoudi, D. Reutzel, M. Van Poucke, C. Steinlein, S. Störkel, U. Claussen, T.J. Robinson, J.A.M. Graves, A. Niveleau, F. Grützner, A. Mujica, N. Nomura, W. Vogel, S. Naumann, M. Hughes, E.C. Akeson, S. Steinemann, I. Nanda, S. Bremer, S. Munsche, S.W. Bremer, P.D. Thomsen, F.J. García-Cozar, D. Prawitt, B.U. Zabel, B. Maurer, A. Astola, T. Liehr, F. Domínguez, M.T. Davisson, H. Hanson, T. Hankeln, L.J. Peelman, M.L. Houck, N. Reissmann, Y. Matsuda, J. Gómez-Márquez, M. Leipoldt, A. Cichutek, K.D. Zang, P. Moens, T. Paiss, D.S. Gallagher, M. Schmid, C.G. Mathew, S.A. Krawetz, F. Piumi, S. Schlaubitz, C. Zühlke, F. Boán, K. Benirschke, M. Yerle, I. Schubert, M. Mende, P.J. Kirby, L. Sánchez, and C. Maier

Subjects
Genetics, Index (economics), Subject (documents), Social science, Biology, Molecular Biology, Genetics (clinical)
Published
2001
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19. Subject Index Vol. 85, 1999

Author

O. Bartsch, M. Vaiman, M.R. Matarazzo, M.P. Hande, L. Vieten, P. Marynen, T.D. Bunch, A.A. Bosma, R.A. Veitia, Marijo Kent-First, J.E. Womack, H. Aburatani, C.R. Lopes, A. Törnsten, H. Kusakabe, P.D. Pearce, N. Tanaka, A. Gelhaus, A. Wagner, H.E. McDermid, G.V.N. Velagaleti, M. Hirai, F. Apiou, P.R. Martens, C. Rogel-Gaillard, T. Ishida, N. Bourgeaux, C. Ottolenghi, L.J. Peelman, H.A. Ansari, M. Svelto, T. Antonevich, Giovanna Grimaldi, M.F. Rothschild, M. Yerle, K. Hashimoto, T. Takahashi, Y. Nakahori, R. Taneja, L. Blottière, Y. Koshizuka, M. Horie, G. Gupta, Y. Itoh, J. Schütte, W. Kreß, C.K. Tuggle, H. Hameister, K. Kikuchi, Mariano Rocchi, J.-C. Amé, P. Chardon, M. Isomura, T. Goldammer, M. Kinebuchi, I. Dunham, A. Aventin, M.D. Bishop, J. Kusuda, N. Werner, D.W. Maher, P. Laurent, P. Slijepcevic, J.M. Perez de la Lastra, T.E. Broad, Y. Hippo, D. Ferbus, D. Beare, D. Michael, G. Goubin, M.R.V. Amarante, H. Hayes, S.A. Tharapel, N. Sato, M.-T. Prospéri, B.P. Chowdhary, R.C. Michaelis, R.S. Danziger, F. Bröcker, A. Billault, A. Eggen, K. Jülicher, K. Kasai, J. Vanselow, M.-C. Bissery, M. Mattheeuws, R.D. Horstmann, M. Rosati, N.K. Rushmere, R. Hong, S. Ikegawa, B.S. Klein, B.P. Morgan, R.S. Wilroy, G. Calamita, M. Hirata, M. Bishop, A. Matsuura, M. Heß, K.E. Teague, C. Spalluto, R. Tanuma, R.M. Schmid, J. Sohal, Annamaria Franzè, A. van Zeveren, A. Zsolnai, T. Kodama, J.F. Taylor, A. Fellous, H. Levéziel, A.T. Tharapel, E.L. Jacobson, B. Dutrillaux, D.B. Zimonjic, D.S. Gallagher, P. Peeters, S.K. Davis, S.E. Long, I. Matsushita, A. Malterer, E. Seemanova, A. Mazzone, S. Liptay, B. Grandchamp, M. Schwerin, S. Nishimura, G. Rappold, M.K. Jacobson, J.D. Burzlaff, P.D. Buchanan, Y. Nakamura, B. Opalka, W. Bardenheuer, M. Vozdová, M.C. Yoshida, T. Voet, M. Van Poucke, R. Fürbass, J.T. Woitach, V. Trichet, C. Mecucci, K. Ried, K. Yamada, N.C. Popescu, A. Chase, P. Pinton, R.M. Brunner, C.L. Keck, C. Guenzi, D. Shkolny, S.S. Thorgeirsson, S. Kubíčková, J. Cools, N.C.P. Cross, J.M. Goldman, C.W. Ernst, G. Marquitan, J. Rubeš, H.-F.S. Sun, C.P. Popescu, and C.H. Laundon

Subjects
Genetics, Index (economics), Subject (documents), Social science, Biology, Molecular Biology, Genetics (clinical)
Published
1999
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20. Contents Vol. 85, 1999

Author

T.D. Bunch, M. Hirai, D. Beare, G. Calamita, K. Hashimoto, M. Mattheeuws, O. Bartsch, A. Aventin, T. Takahashi, R.D. Horstmann, M. Heß, Annamaria Franzè, R. Tanuma, P. Marynen, B. Grandchamp, D. Shkolny, R.A. Veitia, M.P. Hande, H. Kusakabe, C.L. Keck, S.S. Thorgeirsson, A.T. Tharapel, H. Levéziel, N. Sato, M. Svelto, R.S. Danziger, A. Fellous, S. Kubíčková, E. Seemanova, D.W. Maher, Giovanna Grimaldi, Y. Itoh, D.S. Gallagher, P.D. Buchanan, T. Goldammer, S.A. Tharapel, L. Vieten, R.M. Brunner, C. Guenzi, H. Aburatani, B. Opalka, J. Kusuda, M. Bishop, C.R. Lopes, Y. Hippo, M. Kinebuchi, M. Horie, S.E. Long, I. Matsushita, J. Vanselow, T. Kodama, J.F. Taylor, A. Mazzone, J.D. Burzlaff, K. Ried, Y. Nakahori, H.E. McDermid, J.M. Perez de la Lastra, M. Vaiman, A. Törnsten, B. Dutrillaux, S. Liptay, M. Isomura, K. Yamada, M.D. Bishop, B.S. Klein, H.A. Ansari, N.K. Rushmere, R. Hong, S. Ikegawa, P.R. Martens, A. Zsolnai, J. Cools, Y. Nakamura, M.R. Matarazzo, A. Eggen, C. Ottolenghi, Mariano Rocchi, A. van Zeveren, W. Bardenheuer, M. Vozdová, D. Michael, M.C. Yoshida, J.-C. Amé, G. Gupta, C.W. Ernst, L.J. Peelman, N. Bourgeaux, R. Taneja, F. Bröcker, M.-C. Bissery, M. Rosati, H. Hayes, A. Billault, T.E. Broad, N. Werner, G. Marquitan, E.L. Jacobson, W. Kreß, D. Ferbus, M. Hirata, B.P. Chowdhary, M. Schwerin, R.M. Schmid, A.A. Bosma, K. Jülicher, P. Chardon, P. Peeters, K.E. Teague, F. Apiou, V. Trichet, C. Rogel-Gaillard, K. Kasai, A. Matsuura, N.C. Popescu, C. Spalluto, L. Blottière, A. Chase, I. Dunham, D.B. Zimonjic, G. Rappold, C.K. Tuggle, M.R.V. Amarante, N. Tanaka, A. Gelhaus, G.V.N. Velagaleti, T. Ishida, M.F. Rothschild, M.K. Jacobson, M. Van Poucke, N.C.P. Cross, J.M. Goldman, T. Antonevich, J. Schütte, R.C. Michaelis, B.P. Morgan, R.S. Wilroy, S. Nishimura, T. Voet, J.T. Woitach, C. Mecucci, J. Rubeš, H.-F.S. Sun, C.P. Popescu, C.H. Laundon, J.E. Womack, Y. Koshizuka, K. Kikuchi, H. Hameister, S.K. Davis, A. Malterer, P. Slijepcevic, M. Yerle, P. Laurent, G. Goubin, M.-T. Prospéri, R. Fürbass, P. Pinton, J. Sohal, Marijo Kent-First, P.D. Pearce, and A. Wagner

Subjects
Botany, Genetics, Biology, Molecular Biology, Genetics (clinical)
Published
1999
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21. Brief Gene Mapping Reports A / B / C

Author

O.A. Haas, J. Yuan, R.M. Anderson, B. Hoyheim, A. Sanchís, C. Rodellar, S. Floriot, W. Liu, P.C. Cooley, K.H. Howell, J. Ando, N.D. Sumption, A.J. Cuticchia, T. Hayashi, R. Schnabel, A. Dufke, N. Lemieux, F. Piumi, D.G. Papworth, A. Van Zeveren, R.M. Brunner, R.B. Phillips, E.L. Smith, S. Maione, Y. Maeda, U. Claussen, B.F. Koop, M. Kuro-o, S.S. Bhat, T. Benazzou, B. Zhu, M. DesGroseilliers, Z.L. Liu, G.P. Di Meo, L. Biltueva, M. Höglund, R.T.D. Oliver, K.C. Jung, G.B. Hubbard, L.J. Peelman, G.A. Rohrer, N. Mandahl, A. Eggen, Y.J. Lee, M.J. Paape, C.G. Artieri, T. Haaf, D. Incarnato, F. von Eggeling, A. Yoshimura, B.D. Young, H. Hiraiwa, K. Mrasek, A. Ozawa, C.M. Moore, F. Fortin, K.R. Schmidt, Y. Takano, N. Martínez Guardia, P. Zaragoza, H. Barthenschlager, K.M. Brasky, D.D. Bannerman, S.E. Parisotto, W.E. Parris, J.E. Womack, A.K. Srivastava, V.K. Maloney, I. Martín-Burriel, D.L. Stevens, L. Yu, M. Yerle, J.H. Lee, R.D. Hall, C. Ramel, G.W. Silk, A. Hill, M.S. Ashwell, M.J. Wakefield, C. Von Kap-Herr, J.C. Strefford, D. Milan, M. Gross, A. Nakata, S.H.S. Ng, M. Nilsson, F. Mertens, N.J. Foot, H. Suzuki, C.S. Park, M. Rachidi, D.T. Goodhead, K. Ohashi, A. Perucatti, G. Rohrer, K.C. Kim, M.E. Szperka, Y. Yue, L. Granjon, J.A. Rogers, N.S. Thomas, N. Li, T.J. Robinson, M. Van Poucke, V. Volobouev, S. Ladd, S.L. Yu, R. Rasero, K.S. Kim, L. Rodríguez, K. Jacobs, S. Taourit, A. Polityko, L. Iannuzzi, I. Panagopoulos, M. Raveendran, D.J. Driscoll, S. Chen, J. Lyahyai, T. Shimogiri, L.A. Mitchell, D. Soglia, T. Liehr, J. Shipley, A. Weise, T. Goldammer, A. Alsop, Y. Obara, R.D. Kulkarni, E. Lemyre, J.A. Crolla, L. Ordovas, R.G. Danzmann, V.M. Aniskin, R.J. Simensen, P.W. Nathanielsz, H.J. Jung, Y. Ando, R.E. Stevenson, M. Mattheeuws, J.L. Williams, H. Yasue, A.-L. Delezoide, D.I. Jin, V. Trifonov, E.E. Connor, T.M. Lane, W.S. Davidson, P. Sacchi, L. LeRoux, S. Kiuchi, M.M. Leland, J.M. Delabar, R. Roy, C.E. Schwartz, A.M. Khalil, N.E. Schlabritz-Loutsevitch, B.G. Dunn, A. Engelbrecht, B.R. DuPont, J.R. Vermeesch, H. Starke, A.E. Cockwell, C. Lopes, M. Morasch, P. Bass, J. Anderson, R. Rullo, Y. Matsuda, Y.-J. Lu, G. Dobigny, A.G. Terasaki, and P. Gonda

Subjects
Genetics, Gene mapping, Biology, Molecular Biology, Genetics (clinical)
Published
2002
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22. A polymorphic CA-repeat at the porcine transferrin (TF) locus

Author

A. Van Zeveren, M. Van Poucke, Marc Mattheeuws, Luc Peelman, and Patrick Chardon

Subjects
Genetics, chemistry.chemical_classification, 0303 health sciences, 030305 genetics & heredity, Ca repeat, Locus (genetics), General Medicine, Biology, Molecular biology, 03 medical and health sciences, chemistry, Transferrin, Animal Science and Zoology, 030304 developmental biology
Published
1999
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23. Gel electrophoretic comparison of light-induced ascorbic acid oxidase from mustard seedlings and from pumpkin tissue

Author

J. Möller and M. van Poucke

Subjects
chemistry.chemical_classification, Oxidase test, Chromatography, biology, Sinapis, Plant Science, General Medicine, Horticulture, Ascorbic acid, biology.organism_classification, Biochemistry, Isozyme, Electrophoresis, Enzyme, chemistry, Light induced, Molecular Biology, Polyacrylamide gel electrophoresis
Abstract

Ascorbic acid oxidase activity was identified in extracts of Sinapis alba after separation by polyacrylamide gel electrophoresis. No isoenzymes could be detected. In comparison to dark-grown controls, seedlings irradiated continuously with far-red light contain the same enzyme but in greater amount.

Published
1970
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24. 154 Exposure of bulls to heat stress had deleterious effects on embryo development

Author

Daniel de Souza Ramos Angrimani, N. Llamas Luceño, A. Van Soom, M. Van Poucke, Katarzyna Joanna Szymańska, and M. Batlle Perez

Subjects
Reproductive technology, Biology, Insemination, Andrology, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, Genetics, medicine, media_common.cataloged_instance, Blastocyst, European union, Molecular Biology, reproductive and urinary physiology, Fertilisation, 030304 developmental biology, media_common, 0303 health sciences, 030219 obstetrics & reproductive medicine, urogenital system, Embryo culture, Embryo, Sperm, medicine.anatomical_structure, Reproductive Medicine, 13. Climate action, embryonic structures, Animal Science and Zoology, Developmental Biology, Biotechnology
Abstract

Concerns about global climate change reducing animal fertility are arising. Therefore, our objective was to determine the effects of increased environmental temperature on Holstein bulls and its effects on sperm quality and embryo development. Frozen semen samples were obtained from 6 bulls exposed to natural heat stress (HS) in August 2016, compared with a lower temperature (control) in March 2016 (temperature-humidity index of up to 74.5 and 40.6, respectively). We evaluated sperm morphology, embryo development, gene expression, inner cell mass/trophectoderm ratio (ICM/TE), and apoptosis cell ratio of Day-8 blastocysts. Sperm morphology was evaluated using eosin/nigrosin staining. Blastocysts were produced by routine in vitro methods (Wydooghe et al. 2014 Reprod. Fertil. Dev.). Cleavage rates were determined 48h after insemination, and blastocyst rates were determined on Days 7 and 8. Expression of NANOG, SOX2, POU5F1, DNMT (1, 3a, 3b), HSP (A1a, A2, A8, 10, 60, 90), HSF1, IFNT2, H19, SNRPN, IGF2, IGF2R, MEST, PHLDA2, MEG3, MEG9, PEG10 and PLAGL1 were analysed. Total RNA was extracted from Day-8 blastocysts for gene expression analysis using RNeasy Micro Kit (Qiagen, Valencia, CA, USA). Reverse transcription and qPCR were performed with iScript (BioRad, Hercules, CA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (BioRad) in a CFX Connect system (BioRad). Blastocysts were differentially stained (Wydooghe et al. 2011 Anal. Biochem.) and analysed using a Leica TCS-SP8×confocal microscope (Leica Microsystems, Wetzlar, Germany). Data analyses included a GLM procedure and paired-samples Student’s t test (P ≤ 0.05). A normal distribution was verified with a UNIVARIATE procedure and Shapiro-Wilk test. A Wilcoxon signed rank test was used to analyse qPCR data. Detached heads (P=0.006) and coiled tails (P=0.018) were significantly lower in the HS group (4.9 and 0.2%, respectively) compared with the control (5.5 and 0.5%). Moreover, proximal droplets (P=0.051) were lower in the HS group (0.7%) compared with the control (1.3%). Remarkably, cleavage and blastocyst rates at Days 7 and 8 were significantly higher in control (78.4, 19.6 and 29.5%, respectively) compared with HS (75, 14.5 and 23.2%). Early and normal blastocysts were grouped as early stage, whereas expanded, hatching and hatched blastocysts were grouped as advanced stage. There was a significant reduction in the HS group of early stage blastocysts on Day 7 and of advanced stage blastocysts on Days 7 and 8. However, in Day-8 blastocysts, there was no significant difference in gene expression for any target gene. Moreover, there were no significant differences in total number of cells or apoptosis cell ratio in blastocysts. However, the ICM/TE ratio was significantly higher (P=0.021) in control (0.7) compared with HS (0.56). Sperm samples collected in August had reduced fertility compared with those obtained in March. Although fewer sperm abnormalities were present in HS, based on decreased blastocyst rates and ICM/TE ratio in embryos produced with HS semen, we inferred that molecular mechanisms for advanced blastocyst development were affected. However, those mechanisms did not involve our target genes. This work was funded by the European Union, Horizon 2020 Marie Sklodowska-Curie Action, REPBIOTECH 675526.

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