Simultaneous Presence of G1 and G4 Mutations in Growth Differentiation Factor 9 Gene of Iranian Sheep
الموضوعات :S. Eghbalsaied 1 , H. Amini 2 , S. Shahmoradi 3 , M. M. Farahi 4
1 - Young Researchers and Elite Club, Isfahan (Khorasgan) branch, Islamic Azad University, Isfahan, Iran
2 - Young Researchers and Elite Club, Isfahan (Khorasgan) branch, Islamic Azad University, Isfahan, Iran
3 - Young Researchers and Elite Club, Isfahan (Khorasgan) branch, Islamic Azad University, Isfahan, Iran
4 - Young Researchers and Elite Club, Isfahan (Khorasgan) branch, Islamic Azad University, Isfahan, Iran
الکلمات المفتاحية: Iran, Afshari, GDF9, G1, G4, Shal, twin birth,
ملخص المقالة :
The fecundity in sheep has been much interested for producers due to its economical importance. Growth differentiation factor 9 (GDF9) is an autosomal gene with over dominance effect for twosingle nucleotide polymorphisms(SNPs), which resulted in sterility in homozygote mutant ewes and additive effect for three SNPs, without showing sterility event. In the present study, DNA was extracted from whole blood of 19 Iranian Afshari and 8 Shal sheep breeds. The extracted DNA was multiplied several folds for G1, G4 and G8 sites in GDF9 gene through polymerase chain reaction(PCR) technique and was sent for sequencing. Results showed that G1-GDF9 mutation (arginine to histidine shift) was detected in Afshari breed, while G4-GDF9 mutation (glutamic acid to lysine shift) was detected in both Afshari and Shal breeds. The observed SNPs were recorded with very low frequency (<8%) in fertile ewes with twining birth, while infertile ewe did not carry the mutant allele. This might indicate an association between these mutations and twin-births in Iranian breeds. Furthermore, these mutations could not be the reason for ewe infertility. In consistent with the rare frequency of infertility in Iranian sheep, the investigated population lacked the G8 or Thoka mutations, which are the most effective mutations in GDF9 gene.This may be due to lack or rare frequency of this SNP in Iranian sheep flocks. In conclusion, G1- and G4-GDF9 SNPs are segregating with very low frequency in Iranian sheep flocks, so that no homozygous ewe for these SNPs was detected in the current study. The same phenomenon could be the case for other important mutations and the sample size should be very large to rule out presence of such mutations in Iranian sheep breeds.
Akbarpour M., Houshmand M., Ghorashi A. and Hayatgheybi H. (2008). Screening for FecGH mutation of growth differentiation factor 9 gene in Iranian Ghezel sheep population. Int. J. Fertil. Steril. 2, 139-144.
Barzegari A., Atashpaz S., Ghabili K., Nemati Z., Rustaei M. and Azarbaijani R. (2010). Polymorphisms in GDF9 and BMP15 associated with fertility and ovulation rate in Moghani and Ghezel sheep in Iran. Reprod. Domest. Anim. 45, 666-669.
Branet M. (2010). A Note on “Amino Acids”. Department of Chemistry and Biochemistry, Rose-Hulman Institute of Technology.
Davis G.H., Galloway S.M., Ross I.K., Gregan S.M., Ward J., Nimbkar B.V., Gray G.D. and Inounu I. (2002). DNA tests in prolific sheep from eight countries provide new evidence on origin of the Booroola (FecB) mutation. Biol. Reprod. 66, 1869-1874.
Eghbalsaied S., Ghaedi K., Shahmoradi S., Pirestani A., Amini H., Saiedi T.,Nicol L. and McNeilly A. (2012). Presence of SNPs in GDF9 mRNA of Iranian Afshari sheep. Int. J. Fertil. Steril. 5, 225-230.
Fabre S., Pierre A., Mulsant P., Bodin L., Di Pasquale E., Persani L.,Monget P. and Monniaux D. (2006). Regulation of ovulation rate in mammals: contribution of sheep genetic models. Reprod. Biol. Endocrinol. 4, 20-31.
Ghaffari M., Nejati-Javaremi A. and Rahimi G. (2009). Detection of polymorphism in BMPR-IB gene associated with twining in Shal sheep using PCR-RFLP method. Int. J. Agric. Biol. 11, 97-99.
Hanrahan J.P., Gregan 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.
Juengel J.L., Hudson N.L., Heath D.A., Smith P., Reader K.L., Lawrence S.B.,O'Connell A.R., Laitinen M.P., Cranfield M. and Groome N.P. (2002). Growth differentiation factor 9 and bone morphogenetic protein 15 are essential for ovarian follicular development in sheep. Biol. Reprod. 67, 1777-1789.
Juengel J.L., O'Connell A.R., French M.C., Proctor L.E., Wheeler R., Farquhar P.A.,Dodds K.G., Galloway S.M., Johnstone P.D. and Davis G.H. (2011). Identification of a line of sheep carrying a putative autosomal gene increasing ovulation rate in sheep that does not appear to interact with mutations in the transforming growth factor beta superfamily. Biol. Reprod. 85, 113-120.
Kasiriyan M.M., Hafezian S.H. and Hassani N. (2011). Genetic polymorphism BMP15 and GDF9 genes in Sangsari sheep of Iran. Int. J. Gen. Mol. Biol. 3, 31-34.
Moradband F., Rahimi G. and Gholizadeh M. (2011). Association of polymorphisms in fecundity genes of GDF9, BMP15 and BMP15-1B with litter size in Iranian Baluchi sheep. Asian-australas J. Anim. Sci. 24, 1179-1183.
Nicol L., Bishop S.C., Pong-Wong R., Bendixen C., Holm L.E., Rhind S.M. and McNeilly A.S. (2009). Homozygosity for a single base-pair mutation in the oocyte-specific GDF9 gene results in sterility in Thoka sheep. Reproduction. 138, 921-933.
Polley S., De S., Brahma B., Mukherjee A., Vinesh P., Batabyal S., Arora J.S., Pan S., Samanta A.K. and Datta T.K. (2010). Polymorphism of BMPR1B, BMP15 and GDF9 fecundity genes in prolific Garole sheep. Trop. Anim. Health Prod. 42, 985-993.
Sadighi M., Bodensteiner K., Beattie A. and Galloway S. (2002). Genetic mapping of ovine growth differentiation factor 9 (GDF9) to sheep chromosome 5. Anim. Gen. 33, 244-245.
Sambrook J. and Russell D.W. (2001). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory Press, New York.
Våge D.I., Husdal M., Kent M.P., Klemetsdal G. and Boman I.A. (2013). A missense mutation in growth differentiation factor 9 (GDF9) is strongly associated with litter size in sheep. BMC Genet. 2, 14.
Yalcin B. (1979). The Sheep Breeds of Afghanistan, Iran and Turkey.Rome: Food and Agriculture Organization of the United Nations.
Yan C., Wang P., De Mayo J., De Mayo F.J., Elvin J.A., Carino C.,Prasad S.V., Skinner S.S., Dunbar B.S. and Dube J.L. (2001). Synergistic roles of bone morphogenetic protein 15 and growth differentiation factor 9 in ovarian function. Mol. End. 15, 854-866.