NEURONAL-GLIAL INTERACTIONS IN CNS DEVELOPMENT

拟议研究计划的长期目标是开发 分子机制研究的模型系统 大脑发育过程中神经元-神经胶质的相互作用及其后果 这些相互作用对两种细胞类型的分化， 对发育调控基因表达的影响。 拟议项目的短期目标和具体目标涉及 使用细胞培养系统来研究特定的 胚胎神经元的类型，通过递质表型鉴定 （多巴胺能，多巴胺能，GABA能）和单层的成熟 星形胶质细胞、成纤维细胞或胚胎神经胶质细胞（放射状神经胶质， 星形胶质细胞）。 这种神经元-神经胶质细胞相互作用对神经细胞的影响是不确定的。 这些识别出递质的神经元的分化和存活将 生物化学测量（3 H-递质的高亲和力摄取） 和形态学（细胞数量的定量和 免疫细胞化学染色的神经元）。 细胞粘附测定，目前 正在开发中，将用于定量这些神经元的粘附 （用自己的3 H-递质放射性标记）与相同的非神经元 上面描述的单层。 据推测， 偏好可能来自特定人群之间的相互作用， 来自同一个大脑的神经元和未成熟的神经胶质细胞 地区或胎龄。 在未来的实验中，我们希望使用粘附试验来测试可能的 介导这些神经元-神经胶质相互作用的分子，以及细胞培养物， 系统来检查这些相互作用对基因表达的影响， 神经元和神经胶质都使用探针来传递细胞特异性信息。

The long-term objectives of the proposed research program are to develop model systems for the study of molecular mechanisms of specific neuronal-glial interactions during brain development, and the consequences of these interactions for the differentiation of both cell types, including effects on the expression of developmentally regulated genes. The short-term goals and specific aims of the proposed project involve the use of a cell culture system to study the interactions between specific types of embryonic neurons, identified by transmitter phenotype (serotonergic, dopaminergic, GABAergic) and monolayers of mature astrocytes, fibroblasts, or embryonic glial cells (radial glia, astrocytes). The effects of such neuronal-glial interactions on the differentiation and survival of these transmitter-identified neurons will be measured both biochemically (high affinity uptake of 3H-transmitters) and morphologically (quantitation of cell number and morphometry of immunocytochemically stained neurons). A cell adhesion assay, currently under development, will be used to quantitate the adhesion of these neurons (radiolabelled with their own 3H-transmitter) to the same non-neuronal monolayers described above. It is hypothesized that distinct adhesion preferences may emerge from interactions between specific populations of transmitter-identified neurons and immature glial cells from the same brain region or gestational age. In future experiments, we hope to use the adhesion assay to test possible molecules mediating these neuronal-glial interactions, and the cell culture system to examine the effects of these interactions on gene expression in both neurons and glia using probes to cell-specific messages.