权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Cellular physiological bases of sex-specific estrogen actions on the brain

性别特异性雌激素对大脑作用的细胞生理基础

基本信息

批准号：
14370025
负责人：
SAKUMA Yasuo
金额：
$ 9.34万
依托单位：
Nippon Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2005
项目状态：
已结题

项目摘要

Transgenic rats expressing enhanced green fluorescent protein (EGFP) under the control of an estrogen receptor (ER) alpha promoter were generated to tag ERalpha-positive neurons in the brain. Two transgenes, one containing sequences for promoter A and DsRed and the other containing sequences for promoter 0/B and EGFP, were injected simultaneously into Wistar rat zygotes. Twenty-two founders with both transgenes were identified. Ten lines of these founders expressed the EGFP tag in the brains of their first filial generation, whereas none similarly expressed the DsRed tag. In two lines selected for the brightness of the EGFP fluorescence in their brains, tagged cells showed essentially the same patterns. Tagged cells were in the preoptic area (POA), bed nucleus of the stria terminalis (BNST), hypothalamic arcuate nucleus and medial amygdala. ERalpha-immunoreactive neurons were identified in all of these structures by immunohistochemistry. In ovariectomized females, approximately 75% of the EGFP-fluorescent cells in the POA-BNST were immunoreactive for ERalpha. In the POA-BNST, ovariectomy increased the number of EGEP-immunopositive cells and estrogen supplementation reversed this effect, indicating that the promoter 0/B is involved in estrogen-induced downregulation of ERalpha. EGFP was also present in cells in the cerebral cortex and hippocampus, which have not previously been associated with endocrine regulation. Conversely, only a few cells were tagged in the hypothalamic ventromedial nucleus, which contained many ERalpha-immunoreactive neurons. This discrepancy could have arisen as a result of differential promoter usage.

产生在雌激素受体（ER）α启动子控制下表达增强型绿色荧光蛋白（EGFP）的转基因大鼠以标记脑中的ER α阳性神经元。将两种转基因，一种含有启动子A和DsRed的序列，另一种含有启动子0/B和EGFP的序列，同时注射到Wistar大鼠受精卵中。确定了22名具有两种转基因的创始人。这些创始人中的10个品系在其第一代子代的大脑中表达了EGFP标签，而没有一个类似地表达了DsRed标签。在两个选择的EGFP荧光亮度在他们的大脑中，标记的细胞显示基本上相同的模式。标记细胞位于视前区（POA）、终纹床核（BNST）、下丘脑弓状核和内侧杏仁核。通过免疫组织化学在所有这些结构中鉴定出ER α免疫反应神经元。在卵巢切除的女性中，POA-BNST中约75%的EGFP荧光细胞对ER α呈免疫反应性。在POA-BNST中，卵巢切除增加了EGEP免疫阳性细胞的数量，而雌激素补充逆转了这种效应，表明启动子0/B参与了雌激素诱导的ER α下调。EGFP也存在于大脑皮层和海马的细胞中，这些细胞以前与内分泌调节无关。相反，只有少数细胞被标记在下丘脑腹内侧核，其中包含许多ER α免疫反应神经元。这种差异可能是由于不同的启动子使用。