Nature of endothelin binding in the porcine ovary

J. A. Flores, T. A. Winters, J. W. Knight, Johannes D Veldhuis

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

We investigated the nature of endothelin (ET) binding in the porcine ovary. We demonstrated the presence of high affinity (K(d) = 0.72; 95% confidence interval = 0.43-1.1 nM) binding sites for ET-1 in the porcine ovary. The binding capacity for this ET-1-specific binding site was 97 pmol/μg DNA (95% confidence interval = 90-107). Autoradiographic studies showed that putative ET receptors reside in the granulosa cell layer of the maturing Graafian follicle and in the vascular components of the corpora lutea. The relative abundance of ET receptors was greatest in granulosa cells of large antral follicles, whereas ET binding was absent in granulosa cells of preantral follicles and in luteal cells. ET binding by cultured granulosa cells was further characterized by RRA and shown to exhibit a rank order of binding affinities for different ET isopeptides. The observed rank order indicates that the ET receptors present on granulosa cells are of the ET(A) receptor subtype. The radioreceptor studies also indicated that granulosa cells collected from large antral follicles (9-10 mm in diameter) have a greater binding capacity for [125I]ET-1 in culture than granulosa cells collected from smaller follicles. When cultured granulosa cells were exposed to 100 nM ET-1 or 14 nM 12-O-tetradecanoylphorbol 13-acetate overnight, the percentage of specific [125I]ET-1 binding was reduced (12% and 50%, respectively), indicating a down-regulation of the ET receptor by these treatments. In summary, we have characterized the distribution, isopeptide specificity, relative abundance, and down-regulation of putative ovarian endothelin receptors of subtype ET(A) on swine granulosa cells. Such results in conjunction with other available literature strongly suggest that granulosa cells of maturing Graafian follicles are targeted by ET-1. An additional physiological role for ET-1 in the ovary is suggested by the presence of putative ET receptors in the vasculature of the corpus luteum.

Original languageEnglish (US)
Pages (from-to)5014-5019
Number of pages6
JournalEndocrinology
Volume136
Issue number11
StatePublished - 1995
Externally publishedYes

Fingerprint

Granulosa Cells
Endothelins
Ovary
Swine
Endothelin Receptors
Endothelin-1
Ovarian Follicle
Corpus Luteum
Cultured Cells
Down-Regulation
Binding Sites
Confidence Intervals
Luteal Cells
Endothelin A Receptors
Tetradecanoylphorbol Acetate
Blood Vessels

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Flores, J. A., Winters, T. A., Knight, J. W., & Veldhuis, J. D. (1995). Nature of endothelin binding in the porcine ovary. Endocrinology, 136(11), 5014-5019.

Nature of endothelin binding in the porcine ovary. / Flores, J. A.; Winters, T. A.; Knight, J. W.; Veldhuis, Johannes D.

In: Endocrinology, Vol. 136, No. 11, 1995, p. 5014-5019.

Research output: Contribution to journalArticle

Flores, JA, Winters, TA, Knight, JW & Veldhuis, JD 1995, 'Nature of endothelin binding in the porcine ovary', Endocrinology, vol. 136, no. 11, pp. 5014-5019.
Flores JA, Winters TA, Knight JW, Veldhuis JD. Nature of endothelin binding in the porcine ovary. Endocrinology. 1995;136(11):5014-5019.
Flores, J. A. ; Winters, T. A. ; Knight, J. W. ; Veldhuis, Johannes D. / Nature of endothelin binding in the porcine ovary. In: Endocrinology. 1995 ; Vol. 136, No. 11. pp. 5014-5019.
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