REGULATION OF NEURONAL SYNAPTIC COMPONENTS

拟议研究的目标是识别分子 神经元突触的成分，并确定细胞的作用- 细胞相互作用在调节细胞的组装和维持中的作用 这种高度专业化的结构。烟碱型胆碱能突触 雏鸡的副交感睫状神经节的原位 被调查，主要是烟碱型乙酰胆碱 受体(AChR)。将用单抗检测AChRs 抗肌肉和电子器官AChRs的抗体 与神经元AChRs发生交叉反应。Pre-and的作用 神经节后相互作用在建立和维持 表面和超微结构中乙酰胆碱受体的数量和分布 早期发育的神经元中的细胞内池将是 用生化和免疫细胞化学技术测定。 细胞-细胞分裂的调控后果 鸡睫状神经节神经元的相互作用将在 开发的后期，当内部AChR池的大小 似乎大大减少了。对一种不同类型的 非突触胆碱能膜成分，α- 银环蛇毒素结合成分，将与AChRs在 发育中的神经元。亲和纯化的特异性抗体 突触后和细胞外基质成分将用于 识别其他神经元突触成分。细胞的作用--细胞 建立和维持层次和空间的相互作用 神经元表面的成分排列将是 下定决心。活性、神经肽和蛋白的独立作用 神经营养因子对突触和非突触的调节 将评估组件并将其与法规 神经节细胞-细胞相互作用在不同时期的后果 发育阶段。AChRs和其他 去神经后识别的突触成分将用于 检查神经再支配部位的特异性。 失神经神经细胞表面原位和在培养的 超微结构水平。预计独特的监管 参与组装、维护和调节的机制 神经突触的功能将被确定。这些研究 也将增加我们在分子水平上的知识 神经元对损伤的反应和相关的特异性程度 随着神经元突触连接的重建。

The goals of the proposed studies are to identified molecular components of neuronal synapses and to determine the role of cell- cell interactions in regulating the assembly and maintenance of this highly specialized structure. Nicotinic cholinergic synapses of the chick parasympathetic ciliary ganglion in situ will be investigated, with major emphasis on the nicotinic acetylcholine receptor (AChR). AChRs will be detected by using monoclonal antibodies to the AChRs of muscle and electric organ that crossreact with neuronal AChRs. The role of pre- and postganglionic interactions in establishing and maintaining the number and ultrastructural distribution of AChRs in surface and intracellular pools in the early developing neurons will be determined using biochemical and immunocytochemical techniques. The regulatory consequences of the disruption of cell-cell interactions will be examined in chick ciliary ganglion neurons at late stages of development when the size of the internal AChR pool appears to be substantially reduced. The regulation of a distinct non-synaptic cholinergic membrane component, the alpha- bungarotoxin binding component, will be compared to AChRs in the developing neurons. Affinity purified antibodies to specific postsynaptic and extracellular matrix components will be used to identify other neuronal synaptic components. The role of cell-cell interactions in establishing and maintaining the levels and spatial arrangement of the components on the neuronal surface will be determined. The independent roles of activity, neuropeptides and neurotrophic factors in the regulation of synaptic and non-synaptic components will be assessed and compared to the regulatory consequences of cell-cell interactions in the ganglion at various developmental stages. The persistence of AChRs and other identified synaptic components after denervation will be used to examine the specificity of the site of reinnervation on the denervated neuron surface in situ and in culture at the ultrastructural level. It is expected that unique regulatory mechanisms involved in the assembly, maintenance and modulation of function at neuronal synapses will be identified. These studies will also increase our knowledge at a molecular level of the neuronal response to damage and the degree of specificty associated with the reestablishment of neuronal synaptic connections.