DEVELOPMENT OF HYPOTHALAMIC NEURONS IN VITRO

这个项目的目的是增加我们对 遗传和表观遗传的作用 哺乳动物中枢神经元的生长和发育 神经系统 神经元的发育 下丘脑，大脑中参与体内平衡的区域 调控，将遵循组织培养。 游离神经元 将在血清中培养 自由条件使用的方法，我们已经开发，和他们的 生长和发育监测使用定量 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 使用适当的单克隆和单特异性抗体。 这些参数的变化是由于治疗 激素和生长因子以及不同的培养基 将提供证据表明，一个特定的表观遗传影响，这些 治疗;稳定的基因表达模式将提供 基因控制的证据。 培养程序的效果 对神经元存活和发育的影响也将使用 3 H-胸腺嘧啶核苷作为神经元“出生日期”的标记物。 的改变 神经元存活或神经元表型作为神经元存活或神经元表型的函数。 将评价解离的时间。 这些的结果 实验将是一个评价的影响，细胞外的 环境，包括生长因子和激素， 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.