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Effect of FSH and LH hormones on oocyte maturation of buffalo and gene expression analysis of their receptors and Cx43 in maturing oocytes

Published online by Cambridge University Press:  04 February 2010

Alok Pandey
Affiliation:
Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India.
S.C. Gupta
Affiliation:
Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India.
Neelam Gupta*
Affiliation:
Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India.
*
All correspondence to: Neelam Gupta. Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India. Tel: +91 941 6401144. Fax: +91 184 2267654. e-mail: [email protected]

Summary

Follicle stimulating hormone (FSH) and luteinizing hormone (LH) are commonly added to maturation media to improve cumulus expansion known to be a predictor of oocyte maturation. Therefore, effects of various concentrations of FSH (1000 ng/ml), LH (1000 ng/ml) and FSH + LH (1000 ng/ml each) in comparison with control (without FSH + LH) cultured oocytes were investigated. FSH and LH (1000 ng/ml each) induced significantly more cumulus expansion and polar body numbers, as compared with control and treatments of 1000 ng/ml FSH and 1000 ng/ml LH alone. Expression of FSH receptor (r), LHr and Cx43 mRNAs was determined by real-time PCR in cumulus–oocyte complexes (COCs) and denuded oocytes at different maturation times. Expression of all three genes was higher in COCs compared with denuded oocytes, confirming the importance of cumulus cells in oocyte maturation. FSHr and connexin 43 (Cx43) mRNA abundance in both COCs and denuded oocytes was highest at 0–6 h of maturation and decreased subsequently. However, LHr mRNA abundance increased from 6 h up to 24 h of maturation. The study concluded that FSH, LH receptors and Cx43 gene expression regulation is an index related to oocyte maturation.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2010

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References

Bustin, S.A. (2000). Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J. Mol. Endocrinol. 25, 169–93.CrossRefGoogle ScholarPubMed
Eppig, J.J., Wigglesworth, K., Pendola, F. & Hirao, Y. (1997). Murine oocytes suppress expression of luteinizing hormone receptor messenger ribonucleic acid by granulosa cells. Biol. Reprod. 56, 976–84.CrossRefGoogle ScholarPubMed
Fagbohun, C.F. & Downs, S.M. (1990). Maturation of the mouse oocyte–cumulus cell complex: stimulation by Lectins. Biol. Reprod. 42, 413–23.CrossRefGoogle ScholarPubMed
Calder, M.D., Caveney, A.N., Smith, L.C. & Watson, A.J. (2003). Responsiveness of bovine cumulus–oocyte-complexes (COC) to porcine and recombinant human FSH and the effect of COC quality on gonadotropin receptor and Cx43 marker gene mRNAs during maturation in vitro. Reprod. Biol. Endocrinol. 1, 1425.CrossRefGoogle ScholarPubMed
Modina, S., Beretta, M., Lodde, V., Lauria, A. & Luciano, A.M. (2004). Cytoplasmic changes and developmental competence of bovine oocytes cryopreserved without cumulus cells. Eur. J. Histochem. 48, 337–46.Google ScholarPubMed
Shimada, M., Maeda, T. & Terada, T. (2001). Dynamic changes of connexin-43, gap junctional protein, in outer layers of cumulus cells are regulated by PKC and PI 3-kinase during meiotic resumption in porcine oocytes. Biol. Reprod. 64, 1255–63.CrossRefGoogle ScholarPubMed
Shimada, M., Hernandez-Gonzalez, I., Gonzalez-Robayna, I. & Richards, J.S. (2006). Paracrine and autocrine regulation of epidermal growth factor-like factors in cumulus oocyte complexes and granulosa cells: key roles for prostaglandin synthase 2 and progesterone receptor. Mol. Endocrinol. 20, 1352–65.CrossRefGoogle ScholarPubMed
Sirard, M.A., Parrish, J.J., & Ware, C.B., Leibfried-Rutledge, M.L., & First, N.L. (1988). The culture of bovine oocytes to obtain developmentally competent embryos. Biol. Reprod. 39, 546–52.CrossRefGoogle ScholarPubMed