FUNCTIONS OF BETA-1 INTEGRINS IN GROWTH CONE MOTILITY

这项提案的长期目标是为了澄清 整合素及其相关分子在生长锥中的作用 能动性。该提案的重点是α-3整合素和α-3 整合素相关蛋白CD151，跨膜蛋白-4- 超级大家庭。第一个具体目标是研究CD151的相关性。 与α-3整合素和信号分子磷脂酰肌醇 免疫共沉淀法在人NT2N神经元中的4-激酶， 免疫印迹和脂蛋白激酶分析。第二个具体目标是 用抗体封闭技术鉴定α-3整合素和CD151 在NT2N神经元突起和突起生长中的作用。第三 具体目标将解决与NT2N工作的活体相关性 α-3整合素和CD151在大脑皮层神经元中的表达 用免疫组织化学和原位技术研究小鼠神经系统发育 杂交。第四个具体目标将涉及功能 α-3整合素S与CD151的相关性。嵌合体 旨在干扰α-3整合素-CD151结合的分子 将其导入NT2前体细胞。NT2N神经元起源于 转基因细胞将在轴突生长试验中进行测试。这个 该提案中的研究将增加我们对整合素的了解 神经元的表达和功能。了解整合素在 神经突起的生长可能使我们能够利用该功能的某些方面 在修复受损的神经或治疗神经病方面 精神错乱。NT2N神经元是可移植的人类神经元，可能 它们本身在治疗神经系统疾病方面是有用的。

The long term goal for this proposal is aimed at elucidating the functions of integrins and integrin-associated molecules in growth cone motility. The proposal is focused on alpha-3 integrin and an alpha-3 integrin-associated protein, CD151, a member of the transmembrane-4- superfamily. The first specific aim will study the association of CD151 with alpha-3 integrin and the signaling molecule phosphatidylinositol 4-kinase in human NT2N neurons using co-immunoprecipitation, immunoblotting, and lipid kinase assays. The second specific aim will use antibody blockade to characterize alpha-3 integrin and CD151 function in neurite initiation and outgrowth by NT2N neurons. The third specific aim will address the in vivo relevance of the work with NT2N neurons by examining the expression of alpha-3 integrin and CD151 in the developing murine nervous system using immunohistochemistry and in situ hybridization. The fourth specific aim will address the functional relevance of alpha-3 integrin s association with CD151. Chimeric molecules designed to perturb the alpha-3 integrin-CD151 association will be transfected into NT2 precursor cells. NT2N neurons derived from the transfectants will be tested in neurite outgrowth assays. The studies in this proposal will add to our knowledge of integrin expression and function in neurons. Understanding integrin function in neurite outgrowth may allow us to exploit some aspect of that function in the repair of damaged nerves or the treatment of neurological disorders. NT2N neurons are transplantable human neurons that may themselves be useful in the treatment of nervous system diseases.