Abstract
Leptin is an adipocyte-derived hormone/cytokine that influences the physiological control of numerous biological functions and links nutritional status with both neuroendocrine and immune functions. In livestock, variation in the leptin (LEP) gene has been characterized in cattle and pig, but it has not been reported in sheep. In this study, variation in the exon 3 coding sequence of the ovine LEP gene was investigated by polymerase chain reaction–single-strand conformational polymorphism (PCR–SSCP) analysis and DNA sequencing. Five novel SSCP patterns, representing five different sequences, were identified under a combination of two different electrophoresis conditions. Either one or two different sequences were detected in individual sheep and all the sequences identified shared high homology with the LEP sequences from a variety of species, suggesting that these sequences represent alleles of the ovine LEP gene. Four single nucleotide polymorphisms (SNPs) were detected, and three of these resulted in amino acid changes. Variation detected here might have an impact on leptin activity and function.
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References
Friedman, J. M., & Halaas, J. L. (1998). Leptin and the regulation of body weight in mammals. Nature, 395, 763–770. doi:10.1038/27376.
Pelleymounter, M. A., Cullen, M. J., Baker, M. B., Hecht, R., Winters, D., Boone, T., et al. (1995). Effects of the obese gene product on body weight regulation in ob/ob mice. Science, 269, 540–543. doi:10.1126/science.7624776.
La Cava, A., & Matarese, G. (2004). The weight of leptin in immunity. Nature Reviews. Immunology, 4, 371–379. doi:10.1038/nri1350.
Matarese, G., La Cava, A., Sanna, V., Lord, G. M., Lechler, R. I., Fontana, S., et al. (2002). Balancing susceptibility to infection and autoimmunity: a role for leptin? Trends in Immunology, 23, 182–187. doi:10.1016/S1471-4906(02)02188-9.
Zhang, Y., Proenca, R., Maffei, M., Barone, M., Leopold, L., & Friedman, J. M. (1994). Positional cloning of the mouse obese gene and its human homologue. Nature, 372, 425–432. doi:10.1038/372425a0.
Halaas, J. L., Gajiwala, K. S., Maffei, M., Cohen, S. L., Chait, B. T., Rabinowitz, D., et al. (1995). Weight-reducing effects of the plasma protein encoded by the obese gene. Science, 269, 543–546. doi:10.1126/science.7624777.
Hager, J., Clement, K., Francke, S., Dina, C., Raison, J., Lahlou, N., et al. (1998). A polymorphism in the 5′ untranslated region of the human ob gene is associated with low leptin levels. International Journal of Obesity, 22, 200–205. doi:10.1038/sj.ijo.0800567.
van der Lende, T., Te Pas, M. F., Veerkamp, R. F., & Liefers, S. C. (2005). Leptin gene polymorphisms and their phenotypic associations. Vitamins and Hormones, 71, 373–404. doi:10.1016/S0083-6729(05)71013-X.
Lagonigro, R., Wiener, P., Pilla, F., Woolliams, J. A., & Williams, J. L. (2003). A new mutation in the coding region of the bovine leptin gene associated with feed intake. Animal Genetics, 34, 371–374. doi:10.1046/j.1365-2052.2003.01028.x.
Liefers, S. C., te Pas, M. F., Veerkamp, R. F., & van der Lende, T. (2002). Associations between leptin gene polymorphisms and production, live weight, energy balance, feed intake, and fertility in Holstein heifers. Journal of Dairy Science, 85, 1633–1638.
Buchanan, F. C., Van Kessel, A. G., Waldner, C., Christensen, D. A., Laarveld, B., & Schmutz, S. M. (2003). Hot topic: an association between a leptin single nucleotide polymorphism and milk and protein yield. Journal of Dairy Science, 86, 3164–3166.
Schenkel, F. S., Miller, S. P., Ye, X., Moore, S. S., Nkrumah, J. D., Li, C., et al. (2005). Association of single nucleotide polymorphisms in the leptin gene with carcass and meat quality traits of beef cattle. Journal of Animal Science, 83, 2009–2020.
Zhou, H., Hickford, J. G. H., & Fang, Q. (2006). A two-step procedure for extracting genomic DNA from dried blood spots on filter paper for PCR amplification. Analytical Biochemistry, 354, 159–161. doi:10.1016/j.ab.2006.03.042.
Liefers, S. C., te Pas, M. F., Veerkamp, R. F., Chilliard, Y., Delavaud, C., Gerritsen, R., et al. (2003). Association of leptin gene polymorphisms with serum leptin concentration in dairy cows. Mammalian Genome, 14, 657–663. doi:10.1007/s00335-003-2275-y.
Sanguinetti, C. J., Dias Neto, E., & Simpson, A. J. (1994). Rapid silver staining and recovery of PCR products separated on polyacrylamide gels. BioTechniques, 17, 914–921.
Zhou, H., & Hickford, J. G. H. (2008). Clonal polymerase chain reaction-single-strand conformational polymorphism analysis: An effective approach for identifying cloned sequences. Analytical Biochemistry, 378, 111–112. doi:10.1016/j.ab.2008.04.005.
Grasso, P., Leinung, M. C., Ingher, S. P., & Lee, D. W. (1997). In vivo effects of leptin-related synthetic peptides on body weight and food intake in female ob/ob mice: Localization of leptin activity to domains between amino acid residues 106–140. Endocrinology, 138, 1413–1418. doi:10.1210/en.138.4.1413.
Acknowledgements
We thank Q. Fang and S. O. Byun for the technical assistance, and the Gene-Marker Laboratory for financial support.
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Zhou, H., Hickford, J.G.H. & Gong, H. Identification of Allelic Polymorphism in the Ovine Leptin Gene. Mol Biotechnol 41, 22–25 (2009). https://doi.org/10.1007/s12033-008-9090-3
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DOI: https://doi.org/10.1007/s12033-008-9090-3