Hirata S, Osada T, Hirai M, Hagihara K, Kato J
Department of Obstetrics and Gynecology, Yamanashi Medical School, Japan.
J Steroid Biochem Mol Biol. 1992 Mar;41(3-8):583-7. doi: 10.1016/0960-0760(92)90388-y.
Reverse transcription-polymerase chain reaction (RT-PCR)-Southern blotting was employed for biochemical detection and measurement of the estrogen receptor messenger ribonucleic acid (ERmRNA) in various parts of the Wistar strain female rat brain. Total RNA extracted from the tissues was subjected to RT-PCR using the primer set which flanked the part(287bp) of the estrogen binding domain-corresponding region of the rat ERcDNA. It was confirmed that the RT-PCR product corresponded to the part of the ERcDNA by direct nucleotide sequencing of the product. Moreover, the level of ERmRNA could be determined by Southern blotting which was highly sensitive and produced quantitative results. The RT-PCR product of about 290 bp, corresponding in length to the distance between two primers, was generated from RNA of all the tissues examined. The levels of the product were as follows; anterior hypophysis greater than hypothalamus and preoptic area, amygdala much greater than cerebral cortex, cerebellum. These results indicate that ERmRNA is widely distributed in the whole brain, implying some physiological action of estrogen on target and 'non-target' brain regions.
采用逆转录-聚合酶链反应（RT-PCR）-Southern印迹法对Wistar品系雌性大鼠脑不同部位的雌激素受体信使核糖核酸（ERmRNA）进行生化检测和测定。从组织中提取的总RNA使用位于大鼠ERcDNA雌激素结合域对应区域部分（287bp）两侧的引物对进行RT-PCR。通过对产物进行直接核苷酸测序，证实RT-PCR产物对应于ERcDNA的部分。此外，通过高度灵敏且能产生定量结果的Southern印迹法可确定ERmRNA的水平。从所有检测组织的RNA中均产生了约290bp的RT-PCR产物，其长度与两个引物之间的距离相对应。产物水平如下：垂体前叶大于下丘脑和视前区，杏仁核远大于大脑皮层、小脑。这些结果表明ERmRNA广泛分布于全脑，提示雌激素对靶脑区和“非靶”脑区有某种生理作用。
