TRANSGENIC OSTEOBLAST SPECIFIC ER ALPHA GENE EXPRESSION

以前的研究表明，雌激素在 在哺乳动物骨骼发育和维持中的作用。 突然 绝经期雌激素的丢失和随后的骨质流失支持了这一点 假说. 然而，迄今为止的大多数调查涉及 使用性腺去势的啮齿动物模型以诱导减少的 荷尔蒙状态 随着雌激素受体α的发育 敲除小鼠，ERKO，我们建议显示雌激素的直接作用 受体α在骨骼发育和维持中的作用。 我们的具体目标 是克隆成骨细胞特异性的启动子（即，人类 骨钙素启动子）的ER-α cDNA的上游，并将其引入 转基因到ERKO小鼠中。 本提案将调查基本的 成骨细胞特异性启动子驱动的雌激素受体的调节- 通过转染在各种成骨细胞样细胞系中表达α和使用 氯霉素乙酰转移酶，北方和受体结合 测定。 该结构最终将被引入C57 Blk/J6 转基因技术。 造骨细胞的产生- 然后可以使用表达特异性ER-α小鼠来测量 雌激素受体-α在野生型基因中的过度表达 背景，使我们能够了解雌激素在骨骼中的作用 发育和骨质疏松症。 ER-α成骨细胞在骨肉瘤中的表达 ERKO背景将开始让我们解决的作用和行动 ER-α基因在ER-α阴性患者骨稳态中的作用 背景

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.