Effects of forskolin and PGE2 on progesterone secretion by goat luteal cells at early and late stages of corpus luteum

Abstract

The aim of this research was to examine the effects of forskolin and PGE2 on steroid synthesis in goat luteal cells, cultured at early and late corpus luteum. Therefore, the luteal cells removed from both stages of the corpus luteum were cultured with newborn calf serum for the first 18 h. Then the media was changed and different concentrations of forskolin (10, 100 ng/ml) or PGE2 (10, 100 ng/ml) were added to the fresh media for another 96 h. The culture media was replaced every 48 h and the retrieval media was kept frozen at -20 °C, until hormone analysis. Luteal cells treated with forskolin produced between 1.87-13.17 times higher production of progesterone, in a dose-dependent manner, compared to the control at early and late stages of corpus luteum (P<0.05). Lower dose of PGE2 increased the progesterone secretion between 2.19-3.28 times in luteal cells compared to the control groups at the late stage of corpus luteum (P<0.05), but not at early stage. The cells treated with a higher dose of PGE2 had no significant effect (P>0.05) on progesterone synthesis at the early and late phases of goat corpus luteum, in comparison to control groups. As a result, this study in goat luteal cells shows that forskolin promotes progesterone synthesis at the early and late corpus luteum, but PGE2 is only effective in cells treated with a low dose at the late stage.

Keywords

• forskolin
• goat
• Luteal cell culture
• pge2
• progesterone

References

1. Alasbahi RH, Melzig MF (2012): Forskolin and derivatives as tools for studying the role of cAMP. Pharmazie, 67, 5-13.
2. Arikan Ş, Rodway RG (2000): Effects of high density lipoprotein containing high or low beta-carotene concentrations on progesterone production and beta-carotene uptake and depletion by bovine luteal cells. Anim Reprod Sci, 62, 253-263.
3. Arikan S, Kalender H, Simsek O (2016): Effects of dbcAMP on progesterone synthesis by cultured goat luteal cell subpopulations isolated from early and late luteal stage corpora lutea. Anim Reprod, 13, 93-99.
4. Arikan Ş, Yigit AA (2003): Changes in size distribution of goat steroidogenic luteal cells during pregnancy. Small Rumin Res, 47, 227-231.
5. Arikan Ş, Yigit AA (2009): Effects of cholesterol and cAMP on progesterone production in cultured luteal cells isolated from pseudopregnant cat ovaries. Anim Reprod Sci, 115, 238-246.
6. Atmaca N, Arikan S, Essiz D, et al (2018): Effects of mancozeb, metalaxyl and tebuconazole on steroid production by bovine luteal cells in vitro. Environ Toxicol Pharmacol, 59, 114-118.
7. Band V, Kharbanda SM, Murugesan K, et al (1986): Effect of forskolin and phosphodiesterase inhibition on prostacyclin-stimulated steroid production in goat ovarian cell types. Prostaglandins Leukot Med, 22, 111-116.
8. Band V, Kharbanda SM, Murugesan K, et al (1987): Steroid production in vitro by granulosa, theca, and luteal cells from goat ovaries. Biol Reprod, 36, 799-806.

9. Baum MGS, Ahrén KEB (1986): Effects of forskolin, luteinizing hormone and prostaglandin F2α on isolated rat corpora lutea. Eur J Endocrinol, 112, 571-578.
10. Chakravarthy P, Goswami J, Sarmah BC (2005): Monitoring of superovulatory response by serum progesterone assay in goat. Indian Vet J, 82, 741-744.
11. Duffy DM, Dozier BL, Seachord CL (2005): Prostaglandin dehydrogenase and prostaglandin levels in periovulatory follicles: Implications for control of primate ovulation by prostaglandin E2. J Clin Endocrinol Metab, 90, 1021-1027.
12. Feng SM, Almond GW (1998): Effects of LH, prostaglandin E2, 8-bromo-cyclic AMP and forskolin on progesterone secretion by pig luteal cells. J Reprod Fertil, 113, 83-89.
13. Fitz TA, Hoyer PB, Niswender GD (1984): Interactions of prostaglandins with subpopulations of ovine luteal cells. I. Stimulatory effects of prostaglandins E1, E2 and I2. Prostaglandins, 28, 119-126.
14. Fitz TA, Mayan MH, Sawyer HR, et al (1982): Characterisation of two steroidogenic cell types in the ovine corpus luteum. Biol Reprod, 27, 703-711.
15. Gregoraszczuk EL, Michas N (1999): Progesterone and estradiol secretion by porcine luteal cells is influenced by individual and combined treatment with prostaglandins E2 and F2 alpha throughout the estrus cycle. Prostaglandins Other Lipid Mediat, 57, 231-241.
16. Hylka VW, Kaki MK, Dizerega GS (1989): Steroidogenesis of porcine granulosa cells from small and medium-sized follicles: Effects of follicle- stimulating hormone, forskolin, and adenosine 3,'5'-cyclic monophosphate versus phorbol ester. Endocrinology, 124, 1204-1209.
17. Kalender H, Arikan S, Simsek O (2014): The effects of LH on progesterone production by cell subpopulations isolated from early and late luteal phase goat corpora lutea. Turkish J Vet Anim Sci, 38, 433-438.
18. Kowalewski MP, Mason JI, Howie AF, et al (2006): Characterization of the canine 3beta-hydroxysteroid dehydrogenase and its expression in the corpus luteum during diestrus. J Steroid Biochem Mol Biol, 101, 254-262.
19. May KCP, Bobe G, Mueller CJ, et al (2011): Conjugated linoleic acid decreases prostaglandin synthesis in bovine luteal cells in vitro. Mol Reprod Dev, 78, 328-336.
20. Niswender GD, Juengel JL, Silva PJ, et al (2000): Mechanisms controlling the function and life span of the corpus luteum. Physiol Rev, 80, 1-29.
21. O’Shaughnessy PJ, Wathes DC (1985): Role of lipoproteins and de-novo cholesterol synthesis in progesterone production by cultured bovine luteal cells. J Reprod Fertil, 74, 425-432.
22. O’Shea JD (1987): Heterogeneous cell types in the corpus luteum of sheep, goats and cattle. J Reprod Fertil, 34, 71-85.
23. Payne JH, Nicholson T, Cooke RG (1993): Insensitivity of dispersed caprine luteal cells to beta-adrenergic agonists and other putative transmitter substances. Theriogenology, 40, 859-863.
24. Rekawiecki R, Nowik M, Kotwica J (2005): Stimulatory effect of LH, PGE2 and progesterone on StAR protein, cytochrome P450 cholesterol side chain cleavage and 3β-hydroxysteroid dehydrogenase gene expression in bovine luteal cells. Prostaglandins Other Lipid Mediat, 78, 169-184.
25. Richards RG, Gadsby JE, Almond GW (1994): Differential effects of LH and PGE2 on progesterone secretion by small and large porcine luteal cells. J Reprod Fertil, 102, 27-34.
26. Sangha GK, Sharma RK, Guraya SS (2002): Biology of corpus luteum in small ruminants. Small Rumin Res, 43, 53-64.
27. Simsek O, Mihm M (2014): Activity of 3β-hydroxysteroid dehydrogenase associated with progesterone production in bovine granulosa cells cultured under different concentrations of serum, insulin-like growth factor I, and gonadotropin. Turkish J Vet Anim Sci, 38, 358-362.
28. Smith MC (1986): The reproductive anatomy and physiology of the female goat. 577-581. In: DA Morrow (Ed), Current Therapy in Theriogenology. W.B. Saunders Company, Philadelphia.
29. Vega M, Devoto L, Navarro V, et al (1987): In vitro net progesterone production by human corpora lutea: Effects of human chorionic gonadotropin, dibutyryl adenosine 3′, 5′-monophosphate, cholera toxin, and forskolin. J Clin Endocrinol Metab, 65, 747-752.
30. Weems CW, Weems YS, Randel RD (2006): Prostaglandins and reproduction in female farm animals. Vet J, 171, 206-228.
31. Wiesak T, Hunter MG, Foxcroft GR (1992): Effect of prostaglandins on luteal function during early pregnancy in pigs. J Reprod Fertil, 95, 831-840.