TRANSGENIC OSTEOBLAST SPECIFIC ER ALPHA GENE EXPRESSION

此前的研究表明，雌激素对人体健康有重要作用 在哺乳动物骨骼发育和维持中的作用。突然性 绝经后雌激素的丢失和随后的骨丢失支持了这一点 假设。然而，迄今为止的大多数调查都涉及 利用性腺去势啮齿动物模型，以诱导减少 荷尔蒙状态。随着雌激素受体-α的发展 基因敲除小鼠Erko，我们建议展示雌激素的直接作用 骨骼发育和维持中的受体-α。我们的具体目标 是为了克隆成骨细胞特异性启动子(即人类 骨钙素启动子)，并介绍了这一点 转基因到尔科小鼠身上。这项提案将调查基本的 成骨细胞特异性启动子驱动的雌激素受体的调节 不同成骨细胞样细胞系中α基因的表达及应用 氯霉素乙酰转移酶、Northern和受体结合 化验。这种结构最终将被引入C57Blk/J6 通过转基因技术获得了小鼠。成骨细胞的产生- 然后可以使用表达特定ER-α的小鼠来测量 雌激素受体-α在野生型遗传病中的过度表达 背景，使我们能够了解雌激素在骨骼中的作用 发育和骨质疏松。成骨细胞ER-α在骨肉瘤中的表达 ERKO背景将开始允许我们处理角色和行动 ER-α阴性患者的ER-α基因在骨内稳态中的作用 背景资料。

Previous investigations have shown that estrogen plays a significant role in the development and maintenance of bone in mammals. The abrupt loss of estrogen at menopause and subsequent bone loss supports this hypothesis. However, the majority of the investigations to date involve the use of a gonadal castrated rodent model in order to induce a reduced hormone state. With the development of the estrogen receptor-alpha knockout mouse, ERKO, we propose to show the direct role of estrogen receptor-alpha in bone development and maintenance. Our specific aims are to clone the promoter of an osteoblastic-specific (i.e., human osteocalcin promoter) upstream of the ER-alpha cDNA and introduce this transgene into the ERKO mouse. This proposal will investigate the basic regulation of the osteoblast specific promoter-driven estrogen receptor- alpha in various osteoblast-like cell lines by transfection and use of chloramphenical acetyltransferase, Northern, and receptor binding assays. This construct will ultimately be introduced into a C57Blk/J6 mouse by using transgenic technology. The creation of an osteoblast- specific ER-alpha expressing mouse can then be used to measure overexpression of the estrogen receptor-alpha in a wild type genetic background, enabling us to understand estrogen's role in bone development and osteoporosis. The ER-alpha osteoblast expression in an ERKO background would begin to allow us to address the role and action of the ER-alpha gene in bone homeostasis in an ER-alpha negative background.