The rat ovarian phospholipase A2 system: Gene expression, cellular localization, activity characterization, and interleukin-1 dependence

Shahar Kol, Kristiina Ruutiainen-Altman, Izhar Ben-Shlomo, Donna W. Payne, Motomu Ando, Eli Y. Adashi

Research output: Contribution to journalArticle

Abstract

We have previously demonstrated that interleukin-1β (IL-1β), a putative intermediary in the ovulatory process, is a potent stimulator of ovarian PG biosynthesis. In this communication, we examine the possibility that this IL-1 effect reflects in part the induction of arachidonic acid mobilization by phospholipase A2 (PLA2). Molecular probing of whole ovarian material revealed the immature rat ovary to be a site of modest secretory PLA2 (sPLA2) gene expression. However, no change in ovarian sPLA2 gene expression was noted during the periovulatory period. Comparable probing for cytosolic PLA2 (cPLA2) failed to disclose a quantifiable signal. However, in situ hybridization localized both sPLA2 and cPLA2 (sPLA2 > cPLA2) transcripts to the granulosa cell layer of the ovarian follicle. Treatment of cultured whole ovarian dispersates with IL-1β produced significant (P < 0.01) increments in the steady state levels of transcripts corresponding to both sPLA2 (1.7-fold increase) and cPLA2 (5-fold increase), an effect reversed by an IL-1 receptor antagonist, suggesting mediation via a specific IL-1 receptor. Treatment with cycloheximide, a protein synthesis inhibitor, resulted in significant attenuation of the ability of IL-1β to up-regulate sPLA2 and cPLA2 gene expression as well as medium PLA2 activity. Treatment with aminoguanidine, an inhibitor of inducible nitric oxide synthase, led to augmentation of the ability of IL-1β to up-regulate sPLA2 and cPLA2 gene expression as well as medium PLA2 activity. Total cellular PLA2 activity proved time, cell density, and calcium dependent, with an optimal pH of 8.0- 9,0 and K(m) values in the low micromolar range (2-5 μM). Our observations 1) establish the rat ovary as a site of sPLA2 and cPLA2 gene expression, 2) localize the corresponding transcripts to the granulosa cell layer, and 3) establish IL-1β as an up-regulatory agent for ovarian sPLA2 and cPLA2 gene expression as well as for ovarian PLA2 activity. These findings also indicate that the IL-1 effect is 1) receptor mediated, 2) contingent in part upon de novo protein biosynthesis, and 3) inhibited by nitric oxide. These observations support the proposition that PLA2 may be a key component in the IL-1-stimulated biosynthesis of ovarian PGs.

Original languageEnglish (US)
Pages (from-to)322-331
Number of pages10
JournalEndocrinology
Volume138
Issue number1
DOIs
StatePublished - Jan 1 1997

ASJC Scopus subject areas

  • Endocrinology

Fingerprint Dive into the research topics of 'The rat ovarian phospholipase A<sub>2</sub> system: Gene expression, cellular localization, activity characterization, and interleukin-1 dependence'. Together they form a unique fingerprint.

  • Cite this