STUDY OF CNS NEUROPEPTIDES WITH PEPTIDE-TOXIN COMPLEXES

中枢神经肽与肽-毒素复合物的研究

• 批准号: 3078263
• 负责人: STEVEN A REEVES
• 金额: $ 5.78万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3078263
• 关键词: Alzheimer's disease; Huntington's disease; Parkinson's disease; basal ganglia; caudate nucleus; cerebral cortex; chemical conjugate; corpus striatum; diphtheria toxin; histochemistry /cytochemistry; immunochemistry; immunoelectrophoresis; immunological substance; mesencephalon; neural degeneration; neurons; neurotoxins; neurotransmitter metabolism; pituitary gland; somatostatin; spinal cord; substantia nigra; thyrotropin releasing hormone; tissue /cell culture; toxin

The goal of this project is to utilize hybrid molecules between neuropeptides and fragments of diptheria toxin or lectins as neurotoxins or pharmacological antagonists to investigate neuropeptide receptor-containing cells in the central nervous system (CNS) with emphasis on the basal ganglia. Thyrotropin releasing hormone (TRH), substance P and somatostatin will be studied. The rationale for the use of these complexes is to target an otherwise nontoxic fragment of diptheria toxin or a lectin to specific neuropeptide receptor-bearing cells by complexing a toxin to a neuropeptide. This project seeks to determine the conditions under which such hybrids are toxic to CNS neurons in the basal ganglia bearing specific neuropeptide receptors in vivo and in vitro. Using an antibody directed against the diptheria toxin fragment, this project will also attempt to visualize neuropeptide receptors on neurons by immunohistochemistry. Previous work demonstrated receptor-specific toxicity of the TRH-diptheria toxin hybrid in vitro against rat pituitary tumor cells. This project will establish conditions for in vivo toxicity of the TRH-toxin hybrid in spinal cord and pituitary, regions where TRH fibers and receptors are present, by examining morphological changes and receptor visualization after systemic or intrathecal delivery of the complex. Having established the feasibility of this methodology for CNS neurons, the neuropeptides substance P and somatostatin, complexed to toxins, will be evaluated for receptor binding and toxicity in vitro in primary cultures of mesencephalic, striatal or cortical neurons. In vivo analysis of substance P and somatostatin-toxin complexes will be performed in the nigrostriatal system by local intracerebral injections and subsequent changes in nigrostriatal neurotransmitter metabolism and behaviors dependent on the integrity of the nigrostriatal system. This tool may provide a means to a better understanding of the processes responsible for the apparent selective vulnerability of some CNS neurons to premature death in degenerative diseases of the nervous system, especially in the basal ganglia (e.g.: Parkinson's, Huntington's, Alzheimer's), by permitting (1) functional and pharmacological analysis of neuropeptide neurons, (2) characterization of distribution of neuropeptide receptors in postmortem tissue, (3) analysis of the factors which regulate the degenerative response to peptide-toxin hybrids, and (4) models of selective neuron loss.

这个项目的目标是利用杂交分子之间 白喉毒素或凝集素的神经肽和片段作为神经毒素， 药理学拮抗剂，以研究含神经肽受体 中枢神经系统（CNS）中的细胞，重点是基底细胞 神经节 促甲状腺激素释放激素（TRH）、P物质和生长抑素 将被研究。 使用这些复合物的基本原理是靶向 白喉毒素的其它无毒片段或凝集素， 神经肽受体携带细胞通过复合毒素， 神经肽 该项目旨在确定在何种条件下， 这种杂合体对基底神经节中的CNS神经元是有毒的， 在体内和体外的神经肽受体。 使用针对 针对白喉毒素片段，该项目还将尝试 通过免疫组织化学使神经元上的神经肽受体可视化。 先前的工作证明TRH白喉的受体特异性毒性 毒素杂合物在体外对大鼠垂体瘤细胞的杀伤作用。 该项目将 建立TRH-毒素杂合体在脊髓中的体内毒性的条件 脊髓和垂体，TRH纤维和受体存在的区域， 检查全身性给药后的形态学变化和受体可视化 或鞘内递送复合物。 在确定了可行性之后， 在CNS神经元的这种方法中，神经肽P物质和 将评价与毒素复合的生长抑素的受体结合 和毒性在体外原代培养中脑，纹状体或 皮质神经元 P物质和生长抑素毒素的体内分析 将在黑质纹状体系统中由当地 脑内注射和随后的黑质纹状体变化 神经递质代谢和行为依赖于完整的 黑质纹状体系统 这个工具可以提供一种更好地理解这些过程的方法 负责一些中枢神经系统神经元对 神经系统退行性疾病的过早死亡，特别是 在基底神经节（例如： 帕金森氏症、亨廷顿氏症、老年痴呆症），由 允许（1）神经肽的功能和药理学分析 神经元，（2）神经肽受体的分布特征， （3）分析了影响死亡率的因素， 对肽-毒素杂合体的退行性反应，和（4）选择性的 神经元丢失