REGULATION OF THE MURINE CALCITONIN RECEPTOR GENE

鼠降钙素受体基因的调控

• 批准号: 6043238
• 负责人: DEBORAH Lynn GALSON
• 金额: $ 32.05万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6043238
• 关键词: DNA binding protein; RNase protection assay; calcitonin; cell differentiation; dexamethasone; embryonic stem cell; gene targeting; genetic regulation; genetic regulatory element; genetically modified animals; hormone receptor; interleukin 1; laboratory mouse; molecular cloning; osteoclasts; polymerase chain reaction; receptor expression; transcription factor

DESCRIPTION (adapted from the Investigator's abstract): Calcitonin (CT) was originally identified as an endocrine hormone that lowers extracellular calcium by inhibiting osteoclastic bone resorption and enhancing renal calcium excretion. The identification of CT receptors in diverse tissues and cell types and the demonstration of its effects on the developing embryo indicates that CT may serve a more complex and diverse function than its role in regulating homeostasis. The discovery that the calcitonin receptor (CTR) gene is alternatively spliced and that certain CTR isoforms may bind ligands other than CTL provides further insight into the molecular and biochemical basis for this functional diversity. In the proposed studies, the Principal Investigators will 1) Clone and characterize the murine CTR gene and identify the transcription start site(s) in osteoclasts and other mCTR-expressing cell-types. 2) Define the specific and potentially unique regulatory sequences responsible for expression of mCTR expressing cells and tissues and identify the putative transacting transcription factors that bind to these sequences. A variety of experimental approaches will be used to define the factors controlling mCTR expression in osteoclasts and other cell types during development. This will include defining the sequences involved in the regulation of mCTR gene expression by calcitonin (CT) and other hormones (e.g., dexamethasone) or cytokines (e.g., IL-1). It is anticipated that the regulatory factors that control mCTR gene expression in osteoclasts also are involved in the regulation of the expression of other genes that accompany the final stages of osteoclast differentiation and activation. 3) Investigate the developmental regulation of the mCTR gene. The temporal and cell-specific pattern of mCTR gene expression will be documented, and then transgenic mCTR-lacZ reporter mice will be used to analyzed the relevance of the regulatory regions identified in Aim 2 and to define any other sequences that may be important for developmentally correct expression in vivo. 4) Determine the effect of targeted disruption of the mCTR gene on osteoclasts and other mCTR-expression tissues, and on mouse development. The Principal Investigator and her colleagues and her colleagues will generate transgenic mice with a targeted disruption of the mCTR gene. In addition, they will develop ES cell differentiation techniques to generate osteoclasts from E cells for studying the regulation of mCTR gene expression during osteoclast differentiation and development.

描述（改编自研究者摘要）：降钙素（CT） 最初被认为是一种内分泌激素， 通过抑制骨细胞的骨吸收， 增强肾钙排泄。CT受体的鉴定 不同的组织和细胞类型，并证明其对 发育中的胚胎表明，CT可能是一个更复杂和多样的 它的功能比它在调节体内平衡中的作用更重要。的发现 降钙素受体（CTR）基因是可变剪接的， CTR同种型可以结合CTL以外的配体，这提供了对 这种功能多样性的分子和生物化学基础。在 建议的研究，主要研究者将1）克隆和 表征鼠CTR基因并鉴定转录起始点 破骨细胞和其他mCTR表达细胞类型中的位点。2)定义 特异性和潜在独特的调控序列， 表达mCTR的细胞和组织的表达，并鉴定推定的 与这些序列结合的转录因子。各种 实验方法将被用来确定控制因素 在发育过程中破骨细胞和其他细胞类型中的mCTR表达。 这将包括定义参与调控的序列， 降钙素（CT）和其它激素（例如， 地塞米松）或细胞因子（例如，IL-1）。预计该 控制破骨细胞中mCTR基因表达的调节因子也 参与调节其他基因的表达， 伴随破骨细胞分化和活化的最后阶段。 3)研究mCTR基因的发育调控。时间 将记录mCTR基因表达的细胞特异性模式， 然后转基因mCTR-lacZ报告小鼠将用于分析 目标2中确定的监管区域的相关性， 其他可能对发育正确的重要序列 体内表达。4)确定有针对性地破坏 mCTR基因对破骨细胞和其它mCTR表达组织以及小鼠的作用 发展首席研究员和她的同事以及她 他的同事们将产生一种转基因小鼠， mCTR基因。此外，他们将开发ES细胞分化 从E细胞产生破骨细胞的技术，用于研究 破骨细胞分化过程中mCTR基因表达的调控 发展