Developmental Action of Estrogen on Cognitive Substrates

雌激素对认知基质的发育作用

基本信息

批准号：
6610022
负责人：
DOMINIQUE CLAUDE TORAN-ALLERAND
金额：
$ 24.53万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-07-01 至 2005-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): We have compelling evidence in both wild-type (+/+) and estrogen receptor-alpha (ER-alpha) gene-disrupted (-/-) (ERKO) postnatal day- (P7) 7 mouse neocortex and uterus that points strongly to the existence of a novel, plasma-membrane-associated, putative ER that is neither ER-alpha (67 kDa) nor ER-beta (54-64 kDa). The proposed studies are designed to further characterize this putative ER, which I have designated "ER-X." "ER-X" is an about 62-63 kDa protein that: (a) is expressed and enriched in purified caveolar-like microdomains of P7, but not adult, neocortical and uterine plasma-membranes; (b) binds 3H-estradiol with high affinity but with ligand specificities radically different from ER-alpha and ER-beta; (c) exhibits some homology with the ligand binding domain of ER-alpha; (d) appears to mediate estrogen activation of the Mitogen-Activated Protein Kinase (MAPK) cascade; (e) is developmentally regulated; and (f) is up-regulated in adult neocortical tissue surrounding an ischemic stroke. We will use standard methods in a series of complementary and correlative cell biological, biochemical, and molecular biological experiments in developing wild-type and ERKO neocortex, using intact tissue; organotypic cultures; and plasma membrane preparations. These methods will include (i) competitive displacement assays, by comparison with 17betaestradiol, (ii) immunoprecipitation and Western blotting; (iii) in situ hybridization; (iv) immunohistochemistry; and (v) cDNA microarray analysis. The experiments are designed to determine (1) Whether "ER-X" is a multiligand receptor; a G-protein coupled receptor or a receptor tyrosine kinase; and (2) To characterize, quantify and compare the genes regulated by ER-X exposure to 17alpha- and 17beta-estradiol. These studies have profound implications for CNS development and cognitive disorders. The results will provide new information regarding the developmental actions and neuroprotective effects of estrogen. Understanding how estrogen influences development may help explain findings that estrogen exerts effects on higher order cognitive processes, and that estrogen or its deficiency may be risk factors for the development of the sexually dimorphic disorders of cognition; schizophrenia; Turner's and Down's Syndromes; and cognitive deficits associated with AIzheimer's and Parkinson's diseases. Understanding developmental mechanisms involving "ER-X" have considerable clinical significance for therapeutic approaches to cognitive deficits, leading the way to drug development and therapeutic intervention selective for "ER-X" in the brain.

描述（由申请人提供）：我们在野生型 (+/+) 和雌激素受体-α (ER-α) 基因破坏 (-/-) (ERKO) 出生后第 (P7) 7 小鼠中都有令人信服的证据新皮质和子宫强烈表明存在一种新型的、与质膜相关的推定 ER，它既不是 ER-α (67 kDa)，也不是 ER-β (54-64) kDa）。拟议的研究旨在进一步描述这种假定的 ER，我将其命名为“ER-X”。 “ER-X”是约62-63kDa的蛋白质，其：(a)在P7的纯化的小凹状微结构域中表达和富集，但不在成人、新皮质和子宫质膜中表达和富集； (b) 以高亲和力结合 3H-雌二醇，但其配体特异性与 ER-α 和 ER-β 完全不同； (c) 与 ER-α 的配体结合结构域表现出一定的同源性； (d) 似乎介导丝裂原激活蛋白激酶 (MAPK) 级联的雌激素激活； (e) 受到发育调节； (f)在缺血性中风周围的成人新皮质组织中表达上调。我们将使用完整组织开发野生型和 ERKO 新皮质的一系列互补和相关细胞生物学、生化和分子生物学实验中的标准方法；器官型培养物；和质膜制剂。这些方法将包括 (i) 竞争性置换测定，与 17β 雌二醇进行比较，(ii) 免疫沉淀和蛋白质印迹； (iii) 原位杂交； (iv) 免疫组织化学； (v) cDNA 微阵列分析。实验旨在确定（1）“ER-X”是否是多配体受体； G蛋白偶联受体或受体酪氨酸激酶； (2) 表征、量化和比较 ER-X 暴露于 17α-和 17β-雌二醇所调节的基因。这些研究对中枢神经系统发育和认知障碍具有深远的影响。结果将提供有关雌激素的发育作用和神经保护作用的新信息。了解雌激素如何影响发育可能有助于解释雌激素对高级认知过程产生影响的发现，以及雌激素或其缺乏可能是性别二态性认知障碍发生的危险因素；精神分裂症;特纳氏综合症和唐氏综合症；以及与阿尔茨海默病和帕金森病相关的认知缺陷。了解涉及“ER-X”的发育机制对于认知缺陷的治疗方法具有相当大的临床意义，为大脑中“ER-X”选择性的药物开发和治疗干预奠定了基础。