EFFECTS OF ESTROGEN ON OSTEOCLASTS FROM TRANSGENIC MICE

关于直接控制经济增长的因素的信息很少。 破骨细胞或负责破骨细胞形成和形成的分子事件 骨吸收。细胞和分子生物学特性的研究 破骨细胞受到严重阻碍，因为无法分离出 破骨细胞或破骨细胞前体的数量和不可获得性 已建立的破骨细胞系。拟议的研究旨在 允许建立表达完全分化的 转基因小鼠破骨细胞瘤中破骨细胞的表型。 然后，这些细胞系将被用来解决有关 雌激素等促骨因子对破骨细胞功能的影响。 拟议的策略涉及指导癌基因的表达， 例如SV40T抗原，对破骨细胞特异或优先作用 利用编码酒石酸抗性酸的基因的调节区 磷酸酶(TRAP)和降钙素受体(CTR)。转基因小鼠 表达这些杂合转基因有望发展成骨肿瘤 破骨细胞起源，或可能永生化的破骨细胞前体 血液、脾或骨髓。这些永生化的细胞将被放置在 将建立培养和细胞系。细胞系将会是 具有广泛的特征以确定它们的相似程度 破骨细胞或破骨细胞前体，并测试它们的能力 文化上的差异化。此外，细胞系将从 雄性和雌性小鼠都可以进行表型比较。 最后，由于雌激素可能会影响 破骨细胞，将进行研究，以确定 雌激素对这些细胞的生长和分化及其影响 吸收骨骼的能力。此外，雌激素将被注射到 转基因小鼠以确定单核细胞或 多核破骨细胞在体内发生改变。 基于转基因小鼠衍生方法的成功 建立了代表其他稀有细胞类型的分化细胞系， 我们希望拟议的研究将导致 已建立的代表破骨细胞或破骨细胞前体的细胞系。 这样的细胞系将为体外分析提供一种新的系统 关于雌激素和其他促骨作用的重要问题 调节破骨细胞功能的因素或辅助细胞。

Little information is available on factors directly controlling the osteoclast or the molecular events responsible for osteoclast formation and bone resorption. The cell and molecular biological characterization of the osteoclast has been severely hampered by the inability to isolate large numbers of osteoclasts or osteoclast precursors and the unavailability of established osteoclastic cell lines. The proposed studies are designed to allow the establishment of cell lines that express the full differentiated phenotype of osteoclasts from osteoclast tumors formed in transgenic mice. These cell lines will then be used to address questions concerning the effects of estrogen and other osteotropic factors on osteoclast function. The proposed strategy involves directing the expression of an oncogene, such as SV40 Tantigen, specifically or preferentially to osteoclast cells using the regulatory regions of the genes encoding tartrate resistant acid phosphatase (TRAP) and the calcitonin receptor (CTR). Transgenic mice expressing these hybrid transgenes are expected to develop bone tumors of osteoclast origin, or possibly immortalized osteoclast precursors in the blood, spleen or bone marrow. These immortalized cells will be placed in culture and cell lines will be established. The cell lines will be extensively characterized to determine the extent to which they resemble osteoclasts or osteoclast precursors and to test their ability to differentiate in culture. In addition, cell lines will be derived from both male and female mice to allow a comparison of phenotype. Finally, since estrogen may affect the formation and activity of osteoclasts, studies will be undertaken to determine the effects of estrogen on the growth and differentiation of these cells and on their ability to resorb bone. In addition, estrogen will be administered to the transgenic mice to determine if the pattern of mononuclear or multinucleated osteoclastic cells is altered in vivo. Based on the success of the transgenic mouse approach for deriving established differentiated cell lines representing other rare cell types, we are hopeful that the proposed studies will lead to the development of established cell lines representing osteoclasts or osteoclast precursors. Such cell lines would provide a novel system for in vitro analysis of important questions concerning the role of estrogen and other osteotropic factors or accessory cells in regulating osteoclast function.