Endothelin-1 is an autocrine/paracrine regulator of porcine granulosa cells
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We studied whether a novel vasoconstrictor, endothelin-1 (ET-1), is synthesized by and released from porcine granulosa cells, and whether ET-1 acts directly on granulosa cells in an autocrine/paracrine fashion. The dilution curve of the conditioned medium from cultured porcine granulosa cells was parallel to a standard curve of ET-1 in RIA. Reverse-phase HPLC of the conditioned media from the granulosa cells revealed a major peak of ET-1-like immunoreactivity (ET-1-LI) coeluting with standard ET-1. ET-1-LI was released from cultured porcine granulosa cells as a function of time. Northern blot analysis demonstrated the expression of mRNA for prepro-ET-1 in the granulosa cells. We demonstrated the presence of ET-1 in the follicular fluid of porcine ovaries, and that the concentration of ET-1 in large follicles was higher than that in small-medium follicles. ET-1 dose-dependently stimulated cell growth and DNA synthesis in porcine granulosa cells. ET-1 inhibited the FSH- and hCG stimulated accumulation of progesterone in porcine granulosa cells in long-term incubation. These findings suggest that ET-1 produced by porcine granulosa cells may function as an autocrine/paracrine growth factor and modulator of steroidogenesis in ovarian granulosa cells.
Key-wordsEndothelin-1 granulosa cells cell growth DNA synthesis progesterone secretion
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- 5.Koseki C., Imai M., Hirata Y., Yanagisawa M., Masaki T. Autoradiographic distribution in rat tissues of binding sites for endothelin a neuropeptide? Am. J. Physiol. 256: 858, 1989.Google Scholar
- 12.Channing C.P., Ledmitz-Rigby F. Methods for assessing hormone mediated differentiations of ovarian cells in culture and in short term incubations. In: Hardmann J.G., O’Malley B.W. (eds.), Methods in Enzymology, vol. 39. Academic Press, New York, 1975, p.183.Google Scholar
- 13.Patterson Jr. M.K. Measurement of growth and viability of cells in culture. In: Jacoby N.D., Pastan I.H. (eds.) Methods in Enzymology, vol. 58. Academic Press, New York, 1979, p. 141.Google Scholar
- 21.Kamada S., Kubota T., Taguchi M., Aso T. High levels of immunoreactive ęndothelin-1 in human follicular fluids. Hum Reprod. in press 1993.Google Scholar
- 24.Tedeschi C., Hazum E., Kokia E., Ricciarelli E., Adashi E.Y., Payne D.W. Endothelin-1 as a luteinization inhibitor: inhibition of rat granulosa cell progesterone accumulation via selective modulation of key steroidogenic steps affecting both progesterone formation and degradation. Endocrinology 131: 2476, 1992.PubMedGoogle Scholar