Genetic Polymorphism of GDF9 Gene in Iranian Moghani Sheep Breed
محورهای موضوعی : Camel
F. Ala Noshahr
1
(Department of Animal Science, University of Tabriz, Tabriz, Iran)
A. Rafat
2
(Department of Animal Science, University of Tabriz, Tabriz, Iran)
کلید واژه: PCR, polymorphism, GDF9, Moghani sheep,
چکیده مقاله :
The families of TGF-β proteins are the most important growth factors in the ovary for growth and differentiation of early ovarian follicles. Three related oocyte-derived members of the transforming growth factor-β superfamily namely growth differentiation factor 9 (GDF9), BMP15 and BMPR-IB have been shown to be essential for follicular growth and ovulation. Different mutations in the GDF9 gene cause increased ovulation rate and infertility in a dosage-sensitive manner in sheep. In this study, blood samples were collected from 150 Moghani sheep breed using venojects treated with the anti-clot substance (EDTA) and subsequently their DNA content were salted out and extracted. The quantity and quality of extracted DNA was examined using spectrophotometery and gel electrophoresis, respectively. With using a pair of specific primers, the DNA fragment was amplified from exon 1 of GDF9 (462 bp). Digested PCR products with HhaI enzyme showed a G to A substiuation in GDF9 locus. The wild type allele of this gene (G/+) with two restriction site resulted DNA fragments of 156, 52 and 254 bp while the mutant allele (G/-) with one restriction site resulted two DNA fragments with the size of 52 and 410 bp. Genotype frequencies for G (+/+), G (+/-) and G (-/-) were 0.66, 0.24 and 0.1, respectively. From studied luci, GDF9 was polymorphic in Iranian Moghani sheep breed.
خانواده پروتئینهای TGF-B از فاکتورهای مهم رشد و تفرق فولیکولهای تخمدانی میباشند. سه ژن مهم از فوق خانواده B شامل فاکتور 9 (GDF9)، BMP15 و BMPR-IB در رشد و تخمکزایی ضروری میباشند. جهشهای مختلفی در ژن GDF9 سبب افزایش تخمکگذاری و تولید مثل در گوسفند میشود. در این مطالعه نمونههای خونی از 150 گوسفند نژاد مغانی جمعآوری و با افزودن EDTA نگهداری و به آزمایشگاه انتقال یافت. سپس DNA آنها به کمک روش نمکی استخراج گردید. کمیت و کیفیت DNA حاصله به ترتیب توسط روشهای اسپکتوفتومتری و ژل الکتروفورز بررسی شد. توسط یک جفت پرایمر قطعهای از DNA ( bp462) از اگزون ژن GDF9 استحصال گردید. محصولات حاصل از هضم PCR توسط آنزیم HhaI جانشینی باز G به A را در لوکاس GDF9 نشان داد. آلل وحشی این ژن (+/G) با دو جایگاه محدود کننده منجر به تولید سه قطعه DNA 156، 52 و bp 254 و آلل جهش یافته (-/G) با یک جایگاه برش سبب تولید دو قطعه 52 و bp 410 شد. فراوانی ژنوتیپی برای ژنوتیپهای (+/+)G، (-/+)G و (-/-)G به ترتیب 66/0، 24/0 و 1/0 به دست آمد. با توجه به مطالعه صورت گرفته چند شکلی ژن GDF9 در گوسفند نژاد مغانی معنیدار به دست آمد.
Chu M.X., Wang S.C.Y.E. and Fang L. (2004). Association between PCR-SSCP of growth differentiation factor 9 genes and high prolificacy in Small Tail Han sheep. Anim. Biotechnol. 15, 111-120.
Chu M.X., Liu Z.H., Giao C.L., He Y.Q., Fang L., Ye S.C., Chen G.H. and Wang G.Y. (2007). Mutations in BMPR-IB and BMP-15 genes are associated with litter size in Small Tailed Han sheep (Ovis aries). J. Anim. Sci. 85, 324-330.
Davis G.H., McEwan J.C., Fennessy P.F., Dodds K.G. and McNatty K.P. (1992). Infertility due to bilateral ovarian hypoplasia in sheep homozygous (FecXI, FecXI) for the Inverdale prolificacy gene located on the X chromosome. Biol. Reprod. 46, 636-640.
Davis G.H., McEwan J.C., Fennessy P.F., Dodds K.G. and Farquhar P.A. (1991). Evidence for the presence of a gene influencing ovulation rate on the X chromosome of sheep. Biol. Reprod. 44, 620-624.
Davis G.H., Balakrishnan L., Ross I.K., Wilson T., Galloway M., Lumsden B.M., Hanrahan J.P., Mullen M.X., Mao Z., Wang G.L., Zhao Z.S., Robinson J.J., Mavrogenis A.P., Papachristoforou C., Peter C., Baumung R., Cardyn P., Boujenane I., Cockett N.E., Eythorsdottir E., Arranz J.J. and Notter D.R. (2006). Investigation of the Booroola (FecB) and Inverdale (FecXI) mutations in 21 prolific breeds and strains of sheep sampled in 13 countries. Anim. Reprod. 92, 87-96.
Galloway S.M., McNatty K.P., Cambridge L.M., Laitinen M.P. E., Juengel J.L., Jokiranta T.S., McLaren R.J., Luiro K., Dodds K.G., Montgomery G.W., Beattie A.E., Davis G.H. and Ritvos O. (2000). Mutations in an oocytederived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner. Nature. 25, 279-283.
Guan F., Liu S.R., Shi G.Q., Ai J.T., Mao D.G. and Yang L.G. (2006). Polymorphism of FecB gene in nine sheep breeds or strains and its effects on litter size, lamb growth and development. Acta Genetic. Sci. 33, 117-124.
Hanrahan J.P., Gergan S.M., Mulsant P., Mullen M., Davis G.H., Powell R. and Galloway S.M. (2004). Mutations in the genes for oocyte derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare Sheep (Ovis aries). Biol. Reprod. 70, 900-909.
Hsueh A.J., McGee E.A., Hayashi M. and Hsu S.Y. (2000). Hormonal regulation of early follicle development in the rate ovary. Mol. Cell. Endocrinol. 163(1), 95-100.
Javanmard A., Azadzadeh N. and Esmailizadeh A. (2011). Mutations in bone morphogenetic protein 15 and growhdiffirentiation factor 9 genes are associated with increased litter size in fat-tailed sheep breeds.Vet. Res. Commun. 35, 157-167.
Juengel J.L., Hudson N.L., Whiting L. and McNatty K.P. (2004). Effects of immunization against bone morphogenetic protein 15 and growth differentiation factor 9 on ovulation rate, fertilization, and pregnancy in ewes. Biol. Reprod. 70, 557-561.
Laitinen M., Vuojolainen K., Jaatinen R., Ketola I., Aaltonen J., Lehtonen E., Heikinheimo M. and Ritvos O. (1998). A novel growth differentiation factor 9 (GDF9) related factors is co-expressed with GDF9 in mouse oocytes during folliculogenesis. Mech. Dev. 78, 135-140.
Liao W.X., Moore R.K. and Shimasaki S. (2004). Functional and molecular characterization of naturally occurring mutations in the oocytesecreted factors bone morphogenetic protein-15 and growth and differentiation factor-9.J. Boil. Chem. 17, 17391-17396.
McNatty K.P., Jungel J.L., Reader K.L., Lun S., Myllymaa S., Lawrence S.B., Western A., Meerasahib M.F., Mottershead D.G., Groome N.P., Ritvos O. and Laitinen M.P. (1997). Bone morphogenetic protein 15 and growth differentiation factor 9 co-operate to regulate granulose cell function in ruminants. Reprod. 129, 481-487.
McNatty K.P., Moore L.G., Hudson N.L., Quirke L.D. and Lawrence S.B. (2004). The oocyte and its role in regulating ovulation rate: anew paradigm in reproductive biology. Reprod. 128, 379-386.
SAS Institute. (2002). SAS®/STAT Software, Release 6.00. SAS Institute, Inc., Cary, NC.
Tavakkolian J. (2000). An Introduction to Genetic Resources of Native Farm Animals in Iran. Animal Science Research Institute, Kraj, Iran.
