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and ß in the Rat Corpus Luteum of Pregnancy: Regulation by Prolactin and Placental Lactogens1
Department of Physiology and Biophysics (C.M.T., L.Z., S.D., R.K.S., N.S., G.G.), College of Medicine, University of Illinois, Chicago, Illinois 60612; and Department of Physiology (K.S.P., O.-K.P.-S.), University of Kentucky, Lexington, Kentucky 40536
Address all correspondence and requests for reprints to: Dr. Geula Gibori, Department of Physiology and Biophysics (M/C 901), University of Illinois, 835 South Wolcott Avenue, Chicago, Illinois 60612-7342. E-mail: ggibori{at}uic.edu
Estradiol, together with PRL and placental lactogens, regulates
steroidogenesis and cell hypertrophy in the rat corpus luteum of
pregnancy. Although binding experiments have demonstrated the presence
of estrogen-binding sites, no evidence exists as to whether the rat
corpus luteum of pregnancy expresses the estrogen receptor (ER) genes.
In this investigation, we have analyzed the expression of the two ER
genes (ER
and ERß) (by RT-PCR and in situ
hybridization) in the rat corpus luteum, studied their developmental
changes throughout pregnancy, and investigated the regulation of ER
and ERß messenger RNA (mRNA) expression by PRL and placental
lactogens. The RT-PCR studies showed that both ER mRNA species (ER
and ERß) are coexpressed in the rat corpus luteum during pregnancy.
Whereas ER mRNA increased from early pregnancy, reached a maximum at
midpregnancy, and had a remarkable decline before parturition; ERß
mRNA remained constant throughout pregnancy, with a significant decline
at parturition. Examination of ER and ERß mRNA expression at the
cellular level, by in situ hybridization, showed ER
expressed in both follicles and corpus luteum, with maximal expression
at midpregnancy. In parallel with the RT-PCR studies, ERß mRNA was
similarly expressed throughout pregnancy in the corpus luteum, but it
was less abundant when compared with small and growing follicles.
Western blot analysis revealed two ER immunoreactive proteins in the
nuclear fraction obtained from pregnant rat corpus luteum: a 67-kDa
moiety, highly expressed at midpregnancy but barely detectable in early
and late gestation; and a 61-kDa form that remained developmentally
unchanged. Hypo- physectomy, performed early in pregnancy, induced a
sharp decline in ER mRNA expression but a less-marked reduction in
ERß mRNA levels. PRL treatment reverted the inhibition induced by
hypophysectomy in both receptor subtypes. When primary luteinized cells
were used to test the effect of PRL, rat placental lactogen I, and rat
placental lactogen II on the expression of ER and ERß mRNA, all
these lactogenic hormones stimulated both ER mRNA species in a
dose-dependent manner. The regulation of ER mRNA expression was further
evaluated in a luteal cell line, termed GG-CL, which apparently
expresses only the ERß mRNA species. Culture of the GG-CL cells, in
the presence of PRL, resulted in a dose-related up-regulation of ERß
mRNA expression. In addition, PRL treatment enhanced the binding
activity of GG-CL cell nuclear proteins to a classical estrogen
response element. Furthermore, in these cells, estradiol treatment
induced a dose-dependent up-regulation of the mRNA encoding protein
kinase C delta isoform, a well-known estrogen target gene in the corpus
luteum of the pregnant rat.
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