ESTROGEN RECEPTOR IN THE RAT BRAIN AND REPRODUCTION (1992)

建立了兔雌激素受体特异性抗血清（AS）。在大肠杆菌中合成的β -半乳糖苷酶- rer结构的融合蛋白作为抗原。使用菌株。用大鼠子宫细胞质组分免疫印迹法证实其特异性。结合研究表明，AS可以识别大鼠内质网的已占用和未占用形式。新生和成年大鼠脑免疫组化（IHC）研究发现，er神经元位于下丘脑核、杏仁核、中脑中央灰核和外侧habenular核。另一方面，以编码大鼠ER的cRNA为探针的原位杂交（ISH）研究显示，ER mrna在脑内相同位置表达，IHC检测到ER阳性信号。内质网与几种神经肽及相关物质的共定位研究表明，一些内质网阳性神经元的细胞质中含有这些物质。雌激素和黄体酮可改变神经细胞的共定位。电镜观察发现，内质网在细胞核上有明显的信号，但核仁上没有这种信号。胞浆中是否存在内质网信号尚不确定。在新生大鼠面神经核腹内侧，IHC和ISH技术均发现ER阳性信号。由于成年大鼠缺乏该位点的信号，因此这种表达只是短暂的。该表达的生物学意义值得进一步分析。

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.