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Regular Article |
a Center for Biomedical Research, The Population Council, New York, New York 10021
b Department of Animal Sciences, University of Missouri-Columbia, Columbia, Missouri 65211
ABSTRACT
Changes in mRNA expression for estrogen receptor (ERß) in relation to mRNAs for LH receptor (LHr) and cytochrome P450 enzymes were examined in granulosa and theca cells from proestrous rat ovarian follicles. Of the 30 ovaries harvested from 15 adult rats, 24 were processed for in situ hybridization, and the remaining were used for reverse transcription-polymerase chain reaction. Messenger RNAs for ERß, LHr, cytochrome P450 side-chain cleavage enzyme (P450scc), 17
-hydroxylase (P450c17), aromatase (P450arom), and steroidogenic acute regulatory protein (StAR) were localized in cross sections of ovaries by in situ hybridization and quantified in granulosa and theca cell layers by a computer-image analyzing system. Ovarian follicles were classified as healthy or atretic. Healthy follicles were divided into four size groups: very small (40–100 µm), small (101–275 µm), medium (276–450 µm), and large (451–850 µm). Atretic follicles were divided into medium (276–450 µm) or large follicles (451–850 µm). A low level of ERß mRNA expression was first detected in granulosa cells of very small healthy follicles, and the expression increased progressively up to medium-sized follicles. The expression of ERß mRNA was highest (P < 0.01) in medium-sized follicles that was followed by a decrease (P < 0.01) in large follicles. Messenger RNAs for LHr, P450scc, and P450arom were first detected in granulosa cells of medium-sized healthy follicles, while mRNAs for LHr, P450scc, P450c17, and StAR were first detected in theca cells associated with very small follicles. The highest expression of LHr, P450scc, P450c17, P450arom, and StAR was seen in granulosa and/or theca cells of large healthy follicles. In atretic follicles, level of gene expression was relatively low in both granulosa and theca cells. In conclusion, stage-specific expression of ERß mRNA was observed in granulosa cells during follicular development. The increased expression of ERß and a concomitant initiation of LHr, P450scc, and P450arom expression in granulosa cells of medium follicles may signify a role for estrogen in follicular development. Also, a strong correlation between ERß mRNA expression in granulosa cells, and the expression of mRNAs for LHr, P450scc, P450c17, and StAR in theca cells associated with growing follicles suggests a possible role for estrogen in steroidogenesis.
First decision: 17 March 2000.
1 This work was supported by cooperative agreement from the U.S. Agency for International Development (Cooperative Agreement CCP-A-00-94-00013-04).
2 Correspondence: Kalyan Sundaram, Center for Biomedical Research, The Population Council, 1230 York Avenue, New York, NY 10021. FAX: 212 327 7678; kalyan{at}popcbr.rockefeller.edu
3 Current address: Ares-Advanced Technology Inc., 280 Pond Street, Randolph, MA 02368.
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