ESTROGEN RECEPTOR IN THE RAT BRAIN AND REPRODUCTION (1992)

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Specific antiserum (AS) against rat estrogen receptor (ER) was raised in a rabbit. As the antigen, a fusion protein of beta-galactosidase-rER construct synthesized in a Escherichia coli. strain was used. The specificity was confirmed by immunoblotting with cytosol fraction of rat uterus. By binding studies, it was shown that the AS recognizes both occupied and unoccupied forms of the rat ER. Immunohistochemical (IHC) studies on the newborn and adult rat brain revealed that the ER-containing neurons were located in the hypothalamiic and amygdalar nuclei, in the midbrain central gray and lateral habenular nucleus. On the other hand, in situ hybridization (ISH) studies, using cRNA coding rat ER as the probe, revealed expression of ER-mRNA in the same location in the brain, where ER positive signals were detected by IHC. Colocalization studies of the ER with several neuropeptides and related substances revealed that some of the ER positive neurons contained these substances in the cytoplasm. The population of colocalized neurons changes by adiministration of estrogen and progesterone. By electron microscopic observations, significant signals indicating the ER were limited on the cell nuclei, but the nucleoli were devoid of the signals. Presence of the ER signals in the cytoplasma was dubious. In the ventromedial part of the facial nucleus in the newborn rat, the ER positive signals were found both by IHC and ISH technique. Since the signals in this site lacked in the adult rat, this expression is only transient. Biological significance of this expression deserves to further analysis.