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Regular Article |
-Hydroxysteroid Dehydrogenase Messenger RNA Transcription in the Immature Rat Ovary in Response to an Ovulatory Dose of Gonadotropin1
a Department of Biology, Trinity University, San Antonio, Texas 78212
b Department of Obstetrics and Gynecology, Kyoto University School of Medicine, Kyoto 606-8507, Japan
c Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030
ABSTRACT
The ovulatory process in mammals involves a substantial increase in the metabolism of steroids and eicosanoids in response to a surge in LH or to an injection of hCG into experimental animals. This study provides evidence that the ovulatory stimulus causes induction of the gene for 3
-hydroxysteroid dehydrogenase (3-HSD), an enzyme that belongs to several oxidoreductase superfamilies that affect steroid and eicosanoid metabolism. Immature Wistar rats were primed with 10 IU eCG s.c., and 48 h later the 12-h ovulatory process was initiated by 10 IU hCG s.c. Ovarian RNA was extracted at 0, 2, 4, 8, 12, and 24 h after injecting the animals with hCG. The RNA extracts were used for reverse transcription-polymerase chain reaction (PCR) differential display to detect gene expression in the stimulated ovarian tissue. One of the PCR primer sets differentially amplified a cDNA fragment that is 52.3% homologous with a 3-HSD gene in rat liver. Northern analyses revealed that maximum transcription was at 8 h after the animals had been treated with hCG. The Northerns also indicated that the 3-HSD cDNA probe cross-hybridized with as many as six different bands of mRNA on the blots. In situ hybridization localized 3-HSD mRNA in the granulosa and thecal layers of mature follicles and in newly formed corpora lutea at 24 h after the ovulatory stimulus. In conclusion, gene(s) for 3-HSD are transcribed in ovarian follicles in response to an ovulatory dose of gonadotropin. A possible function of the oxidoreductase enzyme that is translated from the 3-HSD mRNA may be to reduce the toxic aldehyde and ketone components of the steroids and eicosanoids that accumulate in the mammalian ovary at the time of ovulation.
First decision: 23 October 2000.
1 This work was supported by National Science Foundation grant #9870793 (L.L.E.), by a grant to support T.U. as a Research Fellow of The Lalor Foundation, Providence, Rhode Island (L.L.E.), and by NIH grant HD-16229 (J.S.R.)
2 Correspondence. FAX: 210 999 7229; lespey{at}trinity.edu
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