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Steroid Regulation of Progesterone Receptor Expression in Cultured Rat Gonadotropes¹

Department of Human Physiology (J.L.T., S.M.V., D.W.W.), School of Medicine, University of California, Davis, California 95616; and Division of Life Sciences (G.S.), Lawrence Berkeley National Laboratory, University of California, Berkeley, California 94720

Address all correspondence and requests for reprints to: Dr. Judith L. Turgeon, Department of Human Physiology, School of Medicine, University of California, Davis, California 95616. E-mail: jlturgeon{at}ucdavis.edu

During the preovulatory period, the pituitary action of progesterone is biphasic, moving from a severalfold augmentation of the gonadotropin release action of GnRH to a suppression of GnRH efficacy, which occurs in rats over a period of about 12 h, but the extent to which these biphasic effects are dependent on alterations in progesterone receptor (PR) expression is not known. To address this, as well as the localization of PR in cultured rat pituitary cells, we used cells from ovariectomized rats cultured ± 0.2 nM E₂ with acute progesterone treatment on day 3. Northern blot of poly(A⁺) RNA extracts showed multiple PR messenger RNA (mRNA) transcripts between 4.8–10.2 kb; E₂ treatment led to a 5- to 6-fold increase in the predominant PR mRNA transcripts (5.1 and 10.1 kb). In the presence of E₂, 200 nM progesterone resulted in a decrease in steady-state PR mRNA levels by 3 h of exposure, with the greatest decrease around 6 h (50% of E₂ control) and recovery by 12 h. Similarly treated pituitary cultures were subjected to dual immunofluorescence staining for LH and PR. In the absence of E₂, PR was undetectable. In the presence of E₂, essentially all LH-positive cells were positive for PR and only 1–2% of PR-immunopositive cells were negative for LH, possibly reflecting FSH-exclusive gonadotropes. PR staining was predominantly nuclear, but 20 nM progesterone led to a gradual increase in cytoplasmic staining, with the nuclear-to-cytoplasmic ratio decreasing to near unity by 9–12 h of exposure. In summary, we show for the first time, that PR colocalizes with LH in cultured female rat pituitary cells and that E₂ induces expression of PR mRNA, as well as PR protein, in rat gonadotropes. In the presence of E₂, progesterone causes a rapid but transient down-regulation of PR message; recovery of PR mRNA is accompanied by an increase in cytoplasmic PR, suggestive of an increase in synthesis. These dynamic changes implicate the gonadotrope PR as having a significant role within the temporal context of the rat preovulatory period.

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