DEVELOPMENT OF HYPOTHALAMIC NEURONS IN VITRO

这个项目的目的是增加我们对 遗传和表观遗传以及表观遗传的影响 哺乳动物中枢神经系统的神经元生长发育 神经系统。大脑皮层神经元的发育 下丘脑，大脑中参与动态平衡的区域 在组织培养中，将遵循规则。分离的神经元 从胚胎大鼠的下丘脑将在血清中培养- 使用我们开发的方法的自由条件，以及它们的 使用量化数据监控生长和发育 形态测量技术。特异性神经元的表达 将监测神经递质受体的表型和 使用适当的单抗和单特异性抗体。 这些参数在治疗后的变化 激素和生长因子以及不同培养底物的影响 将为这些基因的特定表观遗传影响提供证据 治疗；稳定的基因表达模式将提供 基因控制的证据。培养程序的影响 关于神经元的生存和发展，也将探索使用 ~3H-胸腺嘧啶核苷作为神经元“生日”的标志。中的更改 神经元存活率或神经元表型与 将评估解离的时间。这一切的结果是 实验将是对细胞外影响的一种评价 环境，包括生长因子和激素，对 下丘脑神经元发育。

The objective of this project is to increase our understanding of the role of genetic and epigenetic and epigenetic influences on neuronal growth and development in the mammalian central nervous system. The development of neurons of the hypothalamus, a region of the brain involved in homeostatic regulation, will be followed in tissue culture. Dissociated neurons from the embryonic rat hypothalamus will be cultured in serum- free conditions using a method we have developed, and their growth and development monitored using quantitative morphometric techniques. The expression of specific neuronal phenotypes and of neurotransmitter receptors will be monitored using appropriate monoclonal and monospecific antibodies. Alterations in these parameters as a result of treatment with hormones and growth factors and by different culture substrates will provide evidence for a specific epigenetic influence of these treatments; a stable pattern of gene expression will provide evidence for genetic control. The effect of the culture procedure on neuronal survival and development will also be explored using 3H-thymidine as a marker of neuronal "birthdate". Alterations in neuronal survival or neuronal phenotype as a function of the timing of dissociation will be evaluated. The result of these experiments will be an appreciation of effects of the extracellular environment, including growth factors and hormones, on hypothalamic neuronal development.