Journal of Livestock Science and Technologies

Journal of Livestock Science and Technologies

Association of novel polymorphisms in follicle stimulating hormone beta (FSHβ) gene with litter size in Mehraban sheep

Document Type : Research Article (Regular Paper)

Authors
1 Department of Animal Science, Faculty of Agriculture, Bu-Ali Sina University, Hamedan, Iran
2 Department of Animal Sciences, Faculty of Agriculture, Bu-Ali Sina University, Hamedan - IRAN
3 Department of Systems and Synthetic Biology, Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
Abstract
Follicle-stimulating hormone (FSH) is necessary for the hypothalamic-pituitary-gonadal axis, and plays an important role in reproduction by binding to a specific receptor (FSHR) through β-subunit of FSH in the surface of the ovarian granulosa cells. This study aimed to characterize FSH β-subunit gene (FSHβ) polymorphism and its association with litter size (LS) using a sample of 118 Mehraban sheep. Polymerase chain reaction (PCR) was performed to amplify two fragments of 300 bp and 431 bp of the ovine FSHβ gene (Oar_v4.0; Chr 15, NC_019472.2). Polymorphisms in the studied fragments were then explored using single strand conformational polymorphism (SSCP) and DNA sequencing methods. A total of seven single-nucleotide polymorphisms (SNPs), including g.59078564 C>G, g.59078624 T>C, g.59078655 T>C, g.59078691 T>C, g.59078754 C>A, g.59080186 G>C and g.59080365 C>T, were found among the six detected SSCP patterns A to F. Moreover, two novel indel polymorphisms called e.g., g.59078702del8-bp−ins64-bp and g.59078726ins54-bp were identified among the three different SSCP genotypes patterns G to I. We found significant differences on prolificacy categories between SSCP genotypes patterns D, E and F (P < 0.01) that simultaneously represented SNP polymorphisms of g.59078754 C>A, g.59080186 G>C and g.59080365 C>T. Similarly, novel indel polymorphisms revealed a significant difference on prolificacy categories between SSCP genotypes patterns G, H and I (P < 0.05). Our results suggest that the FSHβ is a strong candidate gene to associate with the LS in sheep.
Keywords
Subjects

References
Abdoli, R., Mirhoseini, S.Z., Ghavi Hossein-Zadeh, N., Zamani, P., Gondro, C., 2018. Genome-wide association study to identify genomic regions affecting prolificacy in Lori-Bakhtiari sheep. Animal Genetics 49, 488-491.
Abdoli, R., Zamani, P., Deljou, A., Rezvan, H., 2013. Association of BMPR-1B and GDF9 genes polymorphisms and secondary protein structure changes with reproduction traits in Mehraban ewes. Gene 524, 296-303.
Abdoli, R., Zamani, P., Mirhoseini, S.Z., Ghavi Hossein-Zadeh, N., Nadri, S., 2016. A review on prolificacy genes in sheep. Reproduction in Domestic Animals 51, 631-637.
Ahmadi, A., Afraz, F., Talebi, R., Farahavar, A., Vahidi, S.M.F., 2016. Investigation of GDF9 and BMP15 polymorphisms in mehraban sheep to find the missenses as impact on protein. Iranian Journal of Applied Animal Sciences 6, 863-872.
Ahmadi, A., Mohammadizade, M., Zamani, P., Farah-avar, A., Talebi, R., 2021. Identification of polymorphism in HSP70A1A gene in Mehraban and Romanov sheep breeds using PCR-SSCP technique. Research On Animal Production 12, 169-179.
An, X.P., Han, D., Hou, J.X., Li, G., Wang, Y.N., Li, L., Zhu, G.Q., Wang, J., Song, Y.X., Cao, B.Y., 2010. Polymorphism of exon 2 of FSHβ gene and its relationship with reproduction performance in two goat breeds. Agricultural Sciences in China 9, 880-886.
Bianco, B., Loureiro, F.A., Trevisan, C.M., Peluso, C., Christofolini, D.M., Montagna, E., Laganà, A.S., Barbosa, C.P., 2021. Effects of FSHR and FSHB variants on hormonal profile and reproductive outcomes of infertile women with endometriosis. Frontiers in Endocrinology 12, 1-10.
Gholiadeh, M., Najafi, M., 2017. Association of genetic variation in exon 1 and 3 of FSHB gene with litter size in Baluchi sheep. Research on Animal Production 8, 177-182.
Gholizadeh, M., Rahimi-Mianji, G., Nejati-Javaremi, A., De Koning, D.J., Jonas, E., 2014. Genomewide association study to detect QTL for twinning rate in Baluchi sheep. Journal of Genetics 93, 489-493.
Goodman, R.L., Inskeep, E.K., 2015. Control of the ovarian cycle of the sheep. In: Plant, T.M., Zeleznik, A.J. (Eds.), Knobil and Neill's Physiology of Reproduction (Fourth Edition). Elsevier Inc., San Diego, USA, pp. 1259–1305.   
Gootwine, E., 2020. Invited review: Opportunities for genetic improvement toward higher prolificacy in sheep. Small Ruminant Research 186, 106090.
Hediger, R., Johnson, S.E., Hetzel, D.J.S., 2009. Localization of the beta-subunit of follicle stimulating hormone in cattle and sheep by in situ hybridization. Animal Genetics 22, 237-244.
Janssens, S., Vandepitte, W., Bodin, L., 2004. Genetic parameters for litter size in sheep: Natural versus hormone-induced oestrus. Genetics Selection Evolution 36, 543-562.
Laisk, T., Kukuškina, V., Palmer, D., Laber, S., Chen, C.Y., Ferreira, T., Rahmioglu, N., Zondervan, K., Becker, C., Smoller, J.W., Lippincott, M., Salumets, A., Granne, I., Seminara, S., Neale, B., Mägi, R., Lindgren, C.M., 2018. Large-scale meta-analysis highlights the hypothalamic–pituitary–gonadal axis in the genetic regulation of menstrual cycle length. Human Molecular Genetics 27, 4323-4332.
Lameei, H., Afshari, K.P., Abozar, M., 2015. Identification of different allelic forms in exon 1 of FSHβ in Iranian Baluchi and Naeini sheep breeds by PCR-SSCP. Research Opinions in Animal & Veterinary Sciences  5, 360-365.
Liang, C., Chu, M.X., Zhang, J.H., Liu, W.Z., Fang, L., Ye, S.C., 2006. PCR-SSCP polymorphism of FSHbeta gene and its relationship with prolificacy of Jining grey goats. Hereditary 28, 1071-1077.
Liu, J.J., Ran, X.Q., Li, S., Feng, Y., Wang, J.F., 2009. Polymorphism in the first intron of follicle stimulating hormone beta gene in three Chinese pig breeds and two European pig breeds. Animal Reproduction Science 111, 369-375.
Majd, S.A., Ahmadi, A., Talebi, R., Koohi, P.M., Fabre, S., Qanbari, S., 2019. Polymorphism identification in ovine KISS1R/GPR54 gene among pure and crossbreeds of Iranian sheep. Small Ruminant Research 173, 23-29.  
Matoušek, V., Čermáková, A., Kernerová, N., Králová, P., 2005. Analysis of prolificacy in sows of hyperprolific lines of Large White breed. Czech Journal of Animal Science 50, 155-162.
Mitchell, F., 2012. Male fertility influenced by FSHB and FSHR polymorphisms. Nature Reviews Endocrinology 8, 564.
Mokhtari, M.S., Shahrbabak, M.M., Esmailizadeh, A.K., Shahrbabak, H.M., Gutierrez, J.P., 2014. Pedigree analysis of Iran-Black sheep and inbreeding effects on growth and reproduction traits. Small Ruminant Research 116, 14-20.
Nazifi, N., Rahimi-Mianji, G., Ansari-Pirsarai, Z., 2015. Polymorphism in FSHβ and FSHR genes and their relationship with productive and reproductive performance in Iran Black, Arman and Baluchi sheep breeds. Iranian Journal of Applied Animal Science 5, 361-368.
Nikbin, S., Panandam, J.M., Yaakub, H., Murugaiyah, M., 2018. Association of novel SNPs in gonadotropin genes with sperm quality traits of Boer goats and Boer crosses. Journal of Applied Animal Research 46, 459-466.
Niu, X., Martin, G.B., Liu, W., Henryon, M.A., Ren, K., 2019. Follicle-stimulating hormone (FSHβ) gene polymorphisms and associations with reproductive traits in Rex rabbits. Animal Reproduction Science 207, 36-43.
Nosrati, M., Asadollahpour Nanaei, H., Amiri Ghanatsaman, Z., Esmailizadeh, A., 2019. Whole genome sequence analysis to detect signatures of positive selection for high fecundity in sheep. Reproduction in Domestic Animals 54, 358-364.
Pedersen, L.D., Sørensen, A.C., Berg, P., 2009. Marker-assisted selection can reduce true as well as pedigree-estimated inbreeding. Journal of Dairy Science 92, 2214-2223.
Rull, K., Grigorova, M., Ehrenberg, A., Vaas, P., Sekavin, A., Nõmmemees, D., Adler, M., Hanson, E., Juhanson, P., Laan, M., 2018. FSHB -211 G>T is a major genetic modulator of reproductive physiology and health in childbearing age women. Human Reproduction 33, 954-966.
Sanguinetti, C., 1994. RAPD silver staining and recovery of PCR products separated on polyacrylamide gels. Biotechniques 17, 914-921.
Talebi, R., Ahmadi, A., Afraz, F., Sarry, J., Woloszyn, F., Fabre, S., 2018. Detection of single nucleotide polymorphisms at major prolificacy genes in the Mehraban sheep and association with litter size. Annals of Animal Science 18, 685-698.
Talebi, R., Seighalani, R., Qanbari, S., 2021. A handmade DNA extraction kit using laundry powder; insights on simplicity, cost-efficiency, rapidity, safety and the quality of purified DNA. Animal Biotechnology 32, 388-394.
Trevisan, C.M., De Oliveira, R., Christofolini, D.M., Barbosa, C.P., Bianco, B., 2019. Effects of a polymorphism in the promoter region of the follicle-stimulating hormone subunit beta (FSHB) gene on female reproductive outcomes. Genetic Testing and Molecular Biomarkers 23, 39-44.
Vinet, A., Drouilhet, L., Bodin, L., Mulsant, P., Fabre, S., Phocas, F., 2012. Genetic control of multiple births in low ovulating mammalian species. Mammalian Genome 23, 727-740.
Xu, S.S., Gao, L., Xie, X.L., Ren, Y.L., Shen, Z.Q., Wang, F., Shen, M., Eypórsdóttir, E., Hallsson, J.H., Kiseleva, T., Kantanen, J., Li, M.H., 2018. Genome-wide association analyses highlight the potential for different genetic mechanisms for litter size among sheep breeds. Frontiers in Genetics 9, 1-14.
Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., Madden, T.L., 2012. Primer-BLAST: a tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics 13, 134.
Yeh, F.C., Yang, R.C., Boyle, T., 1999. POPGENE. Version 1.31. Microsoft window-based freeware for population genetic analysis. University of Alberta, Edmonton, AB, Canada.
Zamani, P., Nadri, S., Saffaripour, R., Ahmadi, A., Dashti, F., Abdoli, R., 2015. A new mutation in exon 2 of the bone morphogenetic protein 15 gene is associated with increase in prolificacy of Mehraban and Lori sheep. Tropical Animal Health and Production 47, 855-860.
Zhang, C.Y., Wu, C.J., Zeng, W.B., Huang, K.K., Li, X., Feng, J.H., Wang, D., Hua, G.H., Xu, D.Q., Wen, Q.Y., Yang, L.G., 2011. Polymorphism in exon 3 of follicle stimulating hormone beta (FSHB) subunit gene and its association with litter traits and superovulation in the goat. Small Ruminant Research 96, 53-57.