NEUROPEPTIDES & THEIR RECEPTORS IN BRAIN COATED VESICLES

本项目的主要目标是研究亚细胞转运 神经肽（NP）及其受体在脑组织中的表达。 它 特别是旨在展示特定的传递途径， 通过使用一组途径特异性单克隆抗体， 抗体的 该提案只是寻求两个基本问题的答案 关于NP的神经元再循环机制及其 受体，相对于网格蛋白包被的囊泡。 首先， NP及其受体在网格蛋白包被的亚细胞中的转运 细胞器？ 第二，NP和它们的 受体表达的推定途径特异性网格蛋白包被 囊泡？ 最近开发了一组单克隆抗体， 以其识别假定的 胞吐和胞吞网格蛋白包被的囊泡。 推定的 将通过以下方法分离途径特异性CV亚群： 从全脑灰质进行免疫沉淀测定。 的 将通过以下方法测定沉积物中NP的存在和水平： ELISA、免疫印迹、HPLC和放射免疫测定（RIA）方法。 后一种分子技术将用于建立 NP的分子变体存在于不同的亚细胞 运输亚型，即生长抑素1 - 14与生长抑素1 - 14， 28. 还将测定免疫分离的CV亚群的 NP受体分子的存在。 这些决定将依赖于 受体平衡结合技术的应用 或受体亚型选择性标记配体。 的方法 将允许评估特定NP受体亚型是否表达 在网格蛋白包被的囊泡的特定子集中。 的潜在 研究结果可能与同源和异源的 大脑中的受体相互作用系统。 在待研究的NP和受体系统中，一些具有强的 与阿尔茨海默病、亨廷顿舞蹈病和 帕金森氏症， 在老年痴呆症中， 生长抑素水平下降，但不是GABA，两个共同的- 存在于脑组织中的信使 相反的情况发生在 亨廷顿舞蹈病。 这个悖论可以解决，如果 神经元内存在特异性传递途径， 在特定的疾病状态下有不同的改变。

The main goal of this project is to study the subcellular transport of neuropeptides (NP) and their receptors in brain tissue. It particularly aims to demonstrate transmitter-specific pathways through the use of a panel of pathway-specific monoclonal antibodies. The proposal simply seeks for the answers to two basic questions on the mechanisms of neuronal recycling of NP and their receptor(s), in relation to clathrin coated vesicles. First, are NP and their receptors transported in clathrin coated subcellular organelles? And second, in what molecular form are NP and their receptors expressed in putative pathways-specific clathrin coated vesicles? A panel of monoclonal antibodies has been recently developed and characterized with respect to its ability to recognize putative exocytic and endocytic clathrin coated vesicles. The putative pathway specific CV subpopulations will be isolated by immunoprecipitation assays from whole brain gray matter. The sediments will be assayed for the presence and levels of NP by means of ELISAs, immunoblots, HPLC, and radioimmunoassays (RIA). The latter molecular techniques will be employed to establish the molecular variants of NP present in the diverse subcellular transport subsets, i.e. somatostatin 1-14 versus somatostatin 1- 28. The immunodissected CV subpopulations will also be assayed for the presence of NP receptor molecules. These determinations will rely on the application of equilibrium binding techniques using receptor or receptor-subtype selective labeling ligands. The methodology will allow to assess if specific NP receptor subtypes are expressed in specific subsets of clathrin coated vesicles. The potential findings may be intimately linked to homologous and heterologous receptor interaction systems in the brain. Among the NP and receptor systems to be studied, some bear strong relevance to Alzheimer's disease, Huntington's chorea and Parkinson's disease., In Alzheimer's there is a paradoxical decrease in the levels of somatostatin, but not GABA, two co- existing messengers in brain tissue. The converse occurs in Huntington's chorea. This paradox could be resolved if transmitter-specific pathways existed within a neuron and they are differentially altered in specific disease states.