CELLULAR EVENTS IN BONE REMODELING

骨重塑中的细胞事件

• 批准号: 3219123
• 负责人: ROLAND E BARON
• 金额: $ 27.58万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-08-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3219123
• 关键词: adenosinetriphosphatase; apical membrane; basolateral membrane; brush border membrane; calcium metabolism; cell differentiation; cellular pathology; chickens; electron microscopy; extracellular matrix; gene expression; histochemistry /cytochemistry; hormone regulation /control mechanism; immunochemistry; immunocytochemistry; laboratory rabbit; laboratory rat; liver cells; lysosomes; membrane proteins; molecular biology; monoclonal antibody; nutrition related tag; osteoclasts; osteogenesis imperfecta; osteopetrosis; osteoporosis; periodontium disorder; physiologic bone resorption; renal rickets; renal tubule; secretion; sodium potassium exchanging ATPase; tissue /cell culture; transport proteins

The long term objectives of this application are to better understand the cellular events taking place during the process of bone remodeling. This process is sequential and involves a number of steps and a number of cell types. Among these cell types, the osteoclast is the cell responsible for the extracellular resorption of the bone matrix. This cell is also potentially involved, directly or via its precursors, in the recognition of the matrix to resorb. Similarly, the osteoclasts or their activity if potentially involved in the local activation of bone formation. Most diseases of the skeleton, whether metabolic (osteoporosis, renal osteodystrophy), inherited (osteopetrosis, osteogenesis imperfecta), infectious (Paget, periodontal disease, osteoarthritis) or tumoral in nature, involve an abnormality in the bone remodeling sequence. The osteoclasts playing such an essential role in this process, our research program focuses on the cell and molecular biology of the osteoclast's function and differentiation. The specific aims of this application are: 1/ to further characterize the plasma membrane of the mature osteoclast at the molecular level by: a) further determining the role of the (Na+,K+) ATPase subunits in bone resorption, and b) generating new monoclonal and monospecific polyclonal antibodies, to further investigate the function of this cell. 2/ To better understand the biology of the osteoclast at the cellular level and in particular: a) The mechanisms by which lysosomal enzymes and other potential secretory protein(s) are targeted to the ruffled border membrane: b) The differences between the apical (ruffled-border) and baso- lateral membranes; c) The mechanisms by which the polarity of this cell is established and maintained; d) The role of other ion transport proteins in the mechanisms of the bone resorption. 3/ Further compare the osteoclast to other cells involved in transport and acidification (Gastric parietal cell, renal tubule, hepatocyte) 4/ Further analyse the effects of calcium regulating hormones on the distribution, expression and function of the abovementioned molecules and/or mechanisms in which they are involved. The experimental approach selected in this project involves both in vivo and in vitro studies, at the organ, cellular and subcellular levels and in various species (chicken, rat, rabbit). The methods of procedure include the purification and culture of osteoclasts, the fractionation of their membranes, the development of monoclonal and polyclonal antibodies, the analysis of membrane proteins by SDS-PAGE and immunochemical methods, immunocytochemistry at the light and electron microscopic levels, and the in situ hybridization of selected probes.

此应用程序的长期目标是更好 了解在此过程中发生的蜂窝事件 骨骼重塑。 这个过程是顺序的，涉及一个数字 步骤和许多单元格类型。 在这些细胞类型中， 破骨细胞是负责细胞外吸收的细胞 骨基质。 该单元也可能直接参与 或通过其前体，以识别矩阵的回归。 同样，如果有可能涉及的破骨细胞或其活性 在骨形成的局部激活中。 骨骼的大多数疾病，无论是代谢（骨质疏松症） 肾脏骨营养不良），遗传性（骨质骨异常，成骨发生 Imperfecta），感染性（Paget，牙周疾病，骨关节炎） 或肿瘤本质上涉及骨骼异常 重塑序列。 破骨细胞扮演如此重要的 在此过程中的作用，我们的研究计划着重于细胞和 破骨细胞功能和分化的分子生物学。 本应用程序的具体目的是： 1/进一步表征成熟的质膜 分子水平的破骨细胞： a）进一步确定（Na+，K+）ATPase亚基的作用 在骨吸收中，以及 b）产生新的单克隆和单特异性多克隆 抗体，进一步研究该细胞的功能。 2/更好地了解破骨细胞的生物学 细胞水平，尤其是： a）溶酶体酶和其他的机制 潜在的分泌蛋白针对荷叶边边界 膜： b）顶端（荷叶边）和baso-之间的差异 侧膜； c）该细胞的极性为 建立和维护； d）其他离子转运蛋白在机制中的作用 骨吸收。 3/进一步将破骨细胞与其他参与的细胞进行比较 运输和酸化（胃顶细胞，肾小管，肾小管， 肝细胞） 4/进一步分析钙调节激素对激素对 上述的分布，表达和功能 它们所涉及的分子和/或机制。 该项目中选择的实验方法既涉及 体内和体外研究，在器官，细胞和亚细胞中 水平和各种物种（鸡，大鼠，兔子）。 方法 过程包括破骨细胞的纯化和培养， 膜的分馏，发展 单克隆和多克隆抗体，膜的分析 通过SDS-PAGE和免疫化学方法的蛋白质， 在光和电子显微镜下进行免疫细胞化学 水平，以及所选探针的原位杂交。