Actions of endothelin-1 on swine ovarian (granulosa) cells

J. A. Flores, S. Quyyumi, D. A. Leong, Johannes D Veldhuis

Research output: Contribution to journalArticle

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Abstract

We have investigated the regulatory actions of endothelin-1 (ET-1) on inositol phosphate accumulation, cytosolic free Ca2+ ion concentrations ([Ca2+](i)), and basal and FSH-stimulated progesterone and cAMP accumulation by swine granulosa cells in serum-free cultures. ET-1 induced a rapid stimulation of phosphoinositide hydrolysis in populations of granulosa cells, as inferred by the rapid appearance of soluble inositol polyphosphates in response to ET-1 exposure. At the single cell level, fura-2 videomicroscopy was used to measure [Ca2+](i) in individual granulosa cells. We observed cell-cell variability in the threshold concentration of ET-1 required to induce a rise in [Ca2+](i). More than 75% of granulosa cells responded to maximal doses of ET-1. The following parameters of [Ca2+](i) were influenced by ET-1 concentration: percentage of responding cells, lag time for the onset of response, amplitude, and kinetics of the response. Two types of ET-1-mediated [Ca2+](i) rises were observed. One type exhibited rapid Ca2+ kinetics, reaching at least a 2-fold increase above basal (spike phase) within 1-10 sec and returning to a new steady state (plateau phase) 2 min after onset. The other mode of response had slower [Ca2+](i) kinetics, in which 50 sec or more were required to double [Ca2+](i), which remained at this level throughout the observation period (2.5 min). These responses to ET-1 were specific and were not initiated by vasopressin or tumor necrosis factor-α. In cell population studies using monolayer cultures of swine granulosa cells, ET-1 inhibited FSH-stimulated accumulation of progesterone and cAMP. The ET-1-mediated inhibition of FSH- stimulated accumulation of progesterone required at least 4 h of ET-1 exposure. The ET-1-mediated inhibition of both the FSH-stimulated accumulation of progesterone and cAMP after 24-h incubation was mimicked by an activator of protein kinase-C, phorbol 12-myristate 13-acetate, but not by an inactive phorbol. These observations in either single cells or populations of swine ovarian (granulosa) cells are consistent with a possible regulatory role of an ET-1-activated intracellular signaling pathway involving inositol phosphates, [Ca2+](i), and protein kinase-C in the mammalian granulosa cell.

Original languageEnglish (US)
Pages (from-to)1350-1358
Number of pages9
JournalEndocrinology
Volume131
Issue number3
DOIs
StatePublished - 1992
Externally publishedYes

Fingerprint

Granulosa Cells
Endothelin-1
Swine
Progesterone
Inositol Phosphates
Protein Kinase C
Population
Video Microscopy
Polyphosphates
Inositol
Phosphatidylinositols
Vasopressins
Acetates
Hydrolysis
Tumor Necrosis Factor-alpha

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Flores, J. A., Quyyumi, S., Leong, D. A., & Veldhuis, J. D. (1992). Actions of endothelin-1 on swine ovarian (granulosa) cells. Endocrinology, 131(3), 1350-1358. https://doi.org/10.1210/en.131.3.1350

Actions of endothelin-1 on swine ovarian (granulosa) cells. / Flores, J. A.; Quyyumi, S.; Leong, D. A.; Veldhuis, Johannes D.

In: Endocrinology, Vol. 131, No. 3, 1992, p. 1350-1358.

Research output: Contribution to journalArticle

Flores, JA, Quyyumi, S, Leong, DA & Veldhuis, JD 1992, 'Actions of endothelin-1 on swine ovarian (granulosa) cells', Endocrinology, vol. 131, no. 3, pp. 1350-1358. https://doi.org/10.1210/en.131.3.1350
Flores JA, Quyyumi S, Leong DA, Veldhuis JD. Actions of endothelin-1 on swine ovarian (granulosa) cells. Endocrinology. 1992;131(3):1350-1358. https://doi.org/10.1210/en.131.3.1350
Flores, J. A. ; Quyyumi, S. ; Leong, D. A. ; Veldhuis, Johannes D. / Actions of endothelin-1 on swine ovarian (granulosa) cells. In: Endocrinology. 1992 ; Vol. 131, No. 3. pp. 1350-1358.
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