NEUROPEPTIDES & THEIR RECEPTORS IN BRAIN COATED VESICLES

这个项目的主要目标是研究亚细胞的运输。 神经肽(NP)及其受体在脑组织中的分布。它 尤其是旨在展示特定于递质的通路 通过使用一组通路特异性的单抗 抗体。该提案只是寻求两个基本问题的答案 NP神经元再循环机制及其相关问题的探讨 受体(S)，与笼蛋白包裹的囊泡有关。首先，是 NP及其受体在网状蛋白包被亚细胞中的转运 细胞器？第二，NP和它们的分子形式是什么 受体在可能的通路中表达--特异性胞膜蛋白包被 水泡？ 最近开发出了一组克隆抗体，并 以其识别推定的能力为特征 胞外和胞内包裹的囊泡。推定的 途径特定的CV亚群将通过以下方式分离 全脑灰质的免疫沉淀分析。这个 将通过以下方法分析沉积物中NP的存在和水平 ELISA、免疫印迹、高效液相和放射免疫分析(RIA)的方法。 后一种分子技术将被用于建立 NP的分子变体存在于不同的亚细胞中 运输亚群，即生长抑素1-14与生长抑素1- 28. 还将对免疫解剖的CV亚群进行检测 NP受体分子的存在。这些决定将取决于 论受体平衡结合技术的应用 或受体亚型选择性标记配体。方法论 将允许评估特定的NP受体亚型是否表达 在特定的网状蛋白包被小泡中。潜力 这些发现可能与同源和异源密切相关 大脑中的受体相互作用系统。 在待研究的NP和受体系统中，一些具有很强的 与阿尔茨海默病、亨廷顿舞蹈病和 帕金森氏症，在阿尔茨海默氏症中有一种自相矛盾的 生长抑素水平的下降，但不是GABA的水平，两个共同的 脑组织中现有的信使。相反的情况发生在 亨廷顿舞蹈症。这个悖论可以在以下情况下得到解决 神经元中存在特定于递质的通路，它们 在特定的疾病状态下有不同的变化。

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.