CALCITONIN RECEPTOR GENE EXPRESSION

了解参与调控的细胞和分子机制 生理性和病理性骨吸收， 需要开发一种标记物来识别破骨细胞， 这些细胞与造血前体细胞和其他细胞的区别 多核巨细胞（如异物巨细胞）。履行 为了满足这种需要，我们克隆了编码高亲和力受体的cDNA， 降钙素（CT），调节血管生成的主要肽类激素， 破骨细胞的功能活性。此应用程序的目标是 利用我们对人类CT受体（CTR）结构的了解， 分子探针的可用性来回答以下问题：I. 破骨细胞CTR的结构和功能特性是什么？ 从人骨巨细胞瘤和反向骨巨细胞瘤制备的RNA 转录酶/聚合酶链反应（RT-PCR）和克隆技术 将被用于测序和表征结构和 人破骨细胞型CTR的功能特性。2.是什么 骨吸收细胞在生理和 病理性重塑 是否存在没有CTR的破骨细胞"亚型"， 是什么控制了它们的发展？原位杂交技术和 将使用针对人CTR的抗体的免疫组织化学来 显示表达CTR的分布和特定细胞类型。 特定骨病变中细胞的表征和比较，例如 骨的佩吉特病和其他形式的肉芽肿性病变，其中 巨细胞的表型及其与破骨细胞的关系是 不太清楚的事情将被处理。检查来自 对CT不应的个体将提供对 在这些条件下调节CTR表达的机制。 3. CTR基因调控的细胞和分子机制是什么 破骨细胞分化过程中的表达？我们建议克隆和 表征人CTR基因。为了明确具体的和潜在的 负责CTR表达的独特调控序列， 含有推定启动子区的CTR 5 '区的构建体 CTR基因的一部分将与氯霉素转移酶（CAT）连接， 报告基因，并筛选特定激素的能力，或 使用转染在人单核细胞中诱导CTR表达的细胞因子 细胞系该系统可用于识别潜在的独特 有能力调节CTR基因表达的因子。

To understand the cellular and molecular mechanisms involved in regulation of physiological and pathological bone resorption, there is a critical need to develop a marker that identifies osteoclasts and permits distinction of these cells from hematopoietic precursors and other multinucleated giant cells (e.g. foreign body giant cells). To fulfill this need, we have cloned cDNAs that encode high affinity receptors for calcitonin (CT), the principal peptide hormone that regulates the functional activity of osteoclasts. The goal of this application is to exploit our knowledge of the structure of the human CT receptor (CTR) and the availability of molecular probes to answer the following questions: I. What are the structural and functional properties of the osteoclast CTR? RNA prepared from human giant cell tumors of bone and the reverse transcriptase/polymerase chain reaction (RT-PCR) and cloning techniques will be utilized to sequence and characterize the structural and functional properties of the human osteoclast-type CTR. 2. What is the phenotypic relationship between bone resorbing cells in physiological and pathological remodeling? Are there osteoclast "subtypes" without CTRs and what controls their development? In situ hybridization techniques and immunohistochemistry with antibodies to the human CTR will be used to demonstrate the distribution and specific cell-types expressing the CTR. Characterization and comparison of cells in specific bone lesions, e.g. Paget's disease of bone and other forms of granulomatous lesions in which the phenotype of the giant cells and their relationship to osteoclasts is not well understood will be undertaken. Examination of bone samples from individuals with refractoriness to CT will provide insights into the mechanisms by which CTR expression is regulated in these conditions. 3. What are the cellular and molecular mechanisms regulating CTR gene expression during osteoclast differentiation? We propose to clone and characterize the human CTR gene. To define the specific and potentially unique regulatory sequences responsible for expression of the CTR, constructs of the CTR 5' region containing the putative promoter regions of the CTR gene will be linked to a chloramphenicol transferase (CAT) reporter gene and screened for the capacity of specific hormones or cytokines to induce CTR expression using transfection in human mononuclear cell lines. This system can then be used to identify potentially unique factors that have the capacity to regulate CTR gene expression.