Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.50

  • SJR 0.263

  • Impact Factor 0.4 (2024)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Indian Journal of Animal Research, volume 52 issue 5 (may 2018) : 652-657

Effect of in vitro copper supplementation on granulosa cell estradiol synthesis and associated genes

Ravi Kumar Soni­, P.S.P. Gupta, S. Nandi, S. Mondal, J.R. Ippala, Avantika Mor, A. Mishra, Shiv Kumar Tripathi
1ICAR- National Institute of Animal Nutrition and Physiology, Hosur Main Road, Adugodi, Bengaluru-560030, Karnataka India
Cite article:- Soni­ Kumar Ravi, Gupta P.S.P., Nandi S., Mondal S., Ippala J.R., Mor Avantika, Mishra A., Tripathi Kumar Shiv (2018). Effect of in vitro copper supplementation on granulosa cell estradiol synthesis and associated genes. Indian Journal of Animal Research. 52(5): 652-657. doi: 10.18805/ijar.B-3396.
The study was conducted by supplementing cupric chloride dihydrate to modulate the estradiol synthesis in granulosa cells with a hypothesis of possible use of copper to potentiate or partially replace the hormones for estrus induction / estrus synchronization in future studies. In present study copper at three doses (0.1, 0.5 and 1 mM level in culture medium) were tested to deserve see their effects on in vitro granulosa cell survival, estradiol synthesis and their associated genes of ovarian granulosa cells of goat.There was no effect of copper on the ovarian granulosa cell survival rate. There was a considerable  increase in the estradiol level per ml culture medium basis by 6th day of in vitro culture with the second dose of copper i.e. 0.5 mM, but the increase was non-significant (P>0.05). There was no significant effect of copper on estradiol synthesis when expressed on per 30000 cell basis. Effect of copper (0.1 mM and 0.5 mM) on the mRNA expression of genes of aromatase (CYP19A1) and cyclin D2 (CCND2) was estimated. Copper had significantly (P<0.05) increased the mRNA expression of CCND2 and CYP19A1in ovarian granulosa cells with only one of the two doses tested i.e. 0.5 mM. Hence, copper can be considered as a potential mineral to supplement along with hormones in estrus induction or estrus synchronization protocols to minimize the use of hormones.
  1. Behl, R. and Pandey, R. S. (2002). FSH induced stimulation of catalase activity in goat granulosa cells in vitro. Anim Reprod Sci. 70: 215–221.
  2. Basini, S. and Tamanini, C. (2000). Selenium stimulates estradiol production in bovine granulosa cells: possible involvement of nitric oxide. Domest Anim Endocrinol. 18: 1–17.
  3. Bhardwaj, J.K. and Sharma, P. K. (2011). Changes in trace elements during follicular atresia in goat (Capra hircus) ovary. Biol. Trace Elem. Res. 140: 291-298.
  4. Conti, M., (2002). Specificity of the cyclic adenosine 32 , 52 -monophosphate signal in granulosa cell function. Biol. Reprod. 67: 1653–    1661.
  5. Fitzpatrick, S. L. and Richards, J. S. (1991). Regulation of cytochrome P450 aromatase messenger ribonucleic acid and activity by steroids and gonadotropins in rat granulosa cells. Endocrinol. 129: 1452–1462.
  6. Gupta, P.S.P., Folger, J.K., Rajput, S.K., Lv, L., Yao, J., Ireland, J.J., Smith, G.W., (2014). Regulation and regulatory role of WNT signalling in potentiating FSH action during bovine dominant follicle selection. PLOS ONE. 9, e100201.(doi: 10.1371/journal. pone.0100201) 
  7. Grasselli, F., Ponderato, N., Basini, G., Tamanini, C., (2001). Nitric oxide synthase expression and nitric oxide/cyclic GMP pathway in swine granulosa cells. Domest. Anim. Endocrinol. 20: 241–252.
  8. Kendall, N.R., Marsters, P., Scaramuzzi, R.J., Campbell, B.K., (2003). Expression of lysyl oxidase and effect of copper chloride and ammonium tetrathiomolybdate on bovine ovarian follicle granulosa cells cultured in serum-free media. Reproduction. 125: 657–665.
  9. Michaluk, A. and Kochman, K. (2007). Involvement of copper in female reproduction. Reprod. Biol. 7: 193-205.
  10. Pederson, T., (1970). Follicular kinetics in the ovary of the cyclic mouse. Acta. Endocrinol. (Copenh) 64: 304–323.
  11. Qadeer, I., (2006). Factors affecting in vitro maturation, fertilization, and culture of Sahiwal cow (Bosindicus) follicular oocytes to preimplantation embryo. Ph D thesis, University of Agriculture, Faisalabad.
  12. Roychoudhury, S., Bulla, J., Sirotkin, A.V., Kolesarova, A., (2014). In vitro changes in porcine ovarian granulosa cells induced by copper. J. Environ. Sci. Health, Part A, 49: 625-633.
  13. Sen, A., Bettegowda, A., Jimenez-Krassel, F., Ireland, J.J. Smith, G.W., (2007). Cocaine- and amphetamine-regulated transcript regulation of follicle-stimulating hormone signal transduction in bovine granulosa cells. Endocrinol. 148: 4400–4410.
  14. Sherr, C. J., (1994). G1 phase progression: cycling on cue. Cell 79: 551"555 
  15. Silva, J. M. and Price, C. A. (2002). Insulin and IGF-I are necessary for FSH-induced cytochrome P450 aromatase but not cytochrome P450 side-chain cleavage gene expression in oestrogenic bovine granulosa cells in vitro. J. Endocrinol. 174:499–507.
  16. Simpson, E.R. and Devis, S.R., (2001). Minireview: Aromatase and the Regulation of Estrogen Biosynthesis . Endocrinol. 142: 4589-94.
  17. Steinkampf, M.P., Mendelson, C.R., Simpson, E.R., (1987). Regulation by follicle-stimulating hormone of the synthesis of aromatase cytochrome P-450 in human granulosa cells. Mol. Endocrinol. 1: 465–471
  18. Tilly, J. L. and Tilly, K. I. (1995). Inhibition of oxidative stress mimics the ability of FSH to suppress apoptosis in cultured rat ovarian follicles. Endocrinol 136: 242–252
  19. Vázquez-Armijo, J. F., Rojo, R., López, D., Tinoco, J. L., González, A., Pescador, N., DomínguezVara, I. A. (2011). Trace elements in sheep and goats reproduction: A review Tropical and Subtropical Agroecosystems, 14: 1-13
  20. Wang, C., Huang. Y., Dong, K., Liu, Q., Huo, W. (2008). Effect of copper sulphate on reproductive hormone level during estrous cycle in Simmental heifer. Acta Agriculture Nucleatae Sinica.22: 41-46. 
  21. Wilde, D., (2006). Influence of macro and micro minerals in the peri-parturient period on fertility in dairy cattle. Anim. Reprod. Sci. 96: 240-249.
  22. Yang, M.Y. and Rajamahendran, R. (2000). Morphological and biochemical identiûcation of apoptosis in small, medium and large bovine follicles and the effects of FSH and IGF-1 on spontaneous apoptosis in cultured bovine granulosa cells. Biol. Reprod. 62: 1209–1217.
  23. Yunus, E.U., Mustafa, S., Mustafa, T., Baki, H., (2015). Determination of lead, copper and iron in cosmetics, water, soil and food using polyhydroxybutyrate-B-polydimethylsiloxane preconcentration and flame atomic absorption spectroscopy. Anal. Lett. 48: 1163-1179 

Editorial Board

View all (0)