AN APPLICATION OF IN SITU HYBRIDIZATION AND ANTISENSE OLIGONUCLEOTIDE METHOD TO VISUALIZE AND MANIPULATE GENE EXPRESSION OF STEROID HORMONE RECEPTOR PROTEINS IN THE NERVOUS SYSTEM

应用原位杂交和反义寡核苷酸方法可视化和操纵神经系统中类固醇激素受体蛋白的基因表达

基本信息

批准号：
05454674
负责人：
KAWATA Mitsuhiro
金额：
$ 4.16万
依托单位：
KYOTO PREFECTURAL UNIVERSITY OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

项目摘要

Chromaffin cells were found to contain glucocorticoid receptor-immunoreactivity in their nucleus. When dexamethasone was added into the culture medium, the extension of the processes, intensity of immunoreactivity of MAP-2 were significantly diminished. These results indicated that the differentiation of chromaffin cells to neurons is determined, at least in part, by humoral factor of glucocorticoid through its receptor. The addition of antisense oligos for glucocorticoid receptor cleary caused the significant extension of processes of chromaffin cells, indicating that the protein synthesis of steroid receptor was blocked by the antisense inhibitory action to its mRNA.In the sexually dimorphic area of the hypothalamus, immunocytochemical studies show that estrogen induces a significant changes of numbers of immunoreactive never fibers containing neurotensin, calcitonin-gene related peptide (CGRP), Met-enkephalin, cholecystokinin-8 (CCK-8) and dopamine, indicating that axonal sprouting … More of neurons containing these chemical messengers occur in response to estrogen. The relationship between estrogen-receptor and gene expression of these biochemical messengers in concomitant with the process formation or elongation has been discussed, particularly in the context of male and female differences. Estrogen-receptor was found to localize in the cell nucleus by immunocytochemistry in conjunction with the use of estrogen-receptor antibody. Colchicine treatment blocked immunpositive reaction in the cell nucleus, suggesting that estrogen-receptor protein was synthesized in the cytoplasm followed by the transport to the nucleus through nuclear pore in association with microtubules. Monolayr cultures from male and female rat hypothalami were developed and the differences of neurite extension was investigated. In this point, it should be noted that developmental and estrogen-induced changes in mRNAs coding specific proteins for the cytoskeleton, including tubulin-like molecules, have been indentified in the hypothalamic preoptic area. Less

发现铬蛋白细胞在其核us中含有糖皮质激素受体免疫反应性。当将地塞米松添加到培养基中时，过程的扩展，MAP-2的免疫反应性强度显着降低。这些结果表明，至少部分通过糖皮质激素通过其受体的体液因子来确定铬蛋白细胞对神经元的分化。在糖皮质激素受体清洁中添加反义寡聚引起了铬脂细胞的过程的显着延伸，这表明，抗抑制性作用对其mRNA的蛋白质合成被阻断。在其mRNA中，在其mRNA中。在其性二态性区域，免疫细胞的二型二态性区域均未识别出一个显着的脑膜化变化，这是一个显着变化的变化数量的变化数量，这是偶然变化的变化。神经素，降钙素 - 基因相关的肽（CGRP），Met-enkephalin，胆囊化蛋白-8（CCK-8）和多巴胺，表明轴突发芽……更多含有这些化学成员的神经元发生在对雌激素的响应中。已经讨论了这些生化使者与过程形成或伸长的雌激素受体与基因表达之间的关系，特别是在男性和女性差异的背景下。发现雌激素受体可以通过免疫细胞化学与雌激素受体抗体结合使用。秋水仙碱治疗阻断了细胞核中的免疫反应，表明在细胞质中合成雌激素受体蛋白，然后在与微管相关的核孔中转运到核孔中。开发了来自雄性和雌性大鼠下丘脑的单层培养物，并研究了神经蛋白的扩展的差异。在这一点上，应注意的是，在下丘脑前区域已缩进了细胞骨架的发育和雌激素诱导的MRNA编码特异性蛋白的变化，包括小管蛋白样分子。较少的