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a Department of Biology, Trinity University, San Antonio, Texas 78212
b Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030
c Department of Gynecology and Obstetrics, Kyoto University Medical School, Kyoto, Japan
d Department of Obstetrics and Gynecology, Kumamoto University School of Medicine, Kumamoto, Japan
In this differential-display polymerase chain reaction-based study, four different primer sets generated cDNA fragments of ovarian carbonyl reductase genes that were uniquely expressed during the ovulatory process in eCG-primed immature rats. The temporal pattern of expression of this aldo-keto reductase gene was delineated by extracting ovarian RNA at 0, 2, 4, 8, 12, and 24 h after induction of ovulation via injection of the primed animals with hCG. The results showed that at least four homologous forms of this gene were transcribed during ovulation. Northern blot analyses indicated a 14-fold increase in ovarian mRNA for carbonyl reductase, with expression reaching a peak at 8 h after hCG treatment and then declining to negligible levels during the next 16 h. In situ hybridization revealed that most of the transcription was in the thecal connective tissue of the ovary and was absent from the granulosa layer of ovarian follicles. Treatment of the animals with ovulation-blocking doses of epostane (an inhibitor of progesterone synthesis) or indomethacin (an inhibitor of prostanoid synthesis) did not reduce the expression of ovarian carbonyl reductase. Nevertheless, the temporal pattern of expression of carbonyl reductase after the induction of ovulation suggests that this enzyme activity is at least indirectly associated with the ovulatory process.
1 Supported by NSF grant #9870793 (L.L.E.), by a grant to support T.U. as a Research Fellow of The Lalor Foundation, Providence, RI (L.L.E.), and by NIH grant HD-16229 (J.S.R.). This work was presented in part at the 30th Annual Meeting of The Society for the Study of Reproduction held at Portland, OR, August 2-5, 1997.
2 Correspondence. FAX: 210 999 7229; lespey{at}trinity.edu
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