Mouse Col10a1 Gene Regulation and Skeletal Development

小鼠 Col10a1 基因调控和骨骼发育

• 批准号: 6807836
• 负责人: QIPING ZHENG
• 金额: $ 7.53万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6807836
• 关键词: antibody; cartilage development; chondrocytes; collagen; developmental genetics; gel mobility shift assay; gene expression; genetic regulation; genetic regulatory element; genetically modified animals; hypertrophy; in situ hybridization; laboratory mouse; northern blottings; polymerase chain reaction; transcription factor; transfection

DESCRIPTION (provided by applicant): The specific aim of this proposal is to characterize the cis elements and potential transcriptional determinants that confer the tissue-specific expression of type X collagen during chondrocyte hypertrophy. The long-term objective is to understand the molecular regulation of chondrocyte maturation. Chondrocyte hypertrophy is the terminal stage of chondrocyte differentiation during endochondral ossification of long bone growth and type X collagen is the only known hypertrophic chondrocyte-specific molecular marker. Its deficiency in humans causes Schmid metaphyseal chondrodysplasia (SMCD). Mouse genetic studies show that collagen X deficiency has phenotypic abnormalities partly resemble human SMCD. Until now, cis elements or transcription factors that direct its hypertrophic chondrocyte-specific expression in vivo have not been described. The preliminary data within this proposal show that the 10 kb murine Col10a1 promoter/intron element can direct reporter gene expression throughout the hypertrophic zone. In vivo transgenic, in vitro transfection and in silico analysis of Col10a1 suggests that there are conserved elements within both Col10a1 distal promoter and second intron. We propose to further dissect this 10 kb Col10a1 promoter/intron element and the putative DNA-binding proteins that direct its tissue-specific expression in vivo by using transgenic mice approaches and in vitro by using the MCT cell model of chondrocyte hypertrophy. This is essential for understanding the molecular mechanisms that specify endochondral ossification and that underlie the molecular pathogenesis of skeletal dysplasias like those involving type X collagen.

描述（由申请方提供）：本提案的具体目的是表征软骨细胞肥大期间赋予X型胶原组织特异性表达的顺式元件和潜在转录决定簇。长期目标是了解软骨细胞成熟的分子调控。 软骨细胞肥大是长骨生长的软骨内骨化过程中软骨细胞分化的终末阶段，X型胶原是唯一已知的肥大软骨细胞特异性分子标志物。它在人类中的缺乏导致施密德干骺端软骨发育不良（SMCD）。小鼠遗传学研究表明，胶原X缺乏症具有部分类似于人类SMCD的表型异常。到目前为止，指导其肥大软骨细胞特异性表达的顺式元件或转录因子尚未被描述。初步数据表明，10 kb的小鼠Col 10a 1启动子/内含子元件可以直接报告基因在整个肥大区的表达。Col 10a 1的体内转基因、体外转染和计算机模拟分析表明，Col 10a 1远端启动子和第二内含子中存在保守元件。我们建议进一步解剖这个10 kb的Col 10a 1启动子/内含子元件和推定的DNA结合蛋白，通过使用转基因小鼠的方法和在体外通过使用软骨细胞肥大的MCT细胞模型来指导其在体内的组织特异性表达。这对于理解指定软骨内骨化的分子机制以及骨骼发育不良（如涉及X型胶原的骨骼发育不良）的分子发病机制至关重要。