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: 3078262
• 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/3078262
• 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.

该项目的目的是利用杂交分子 神经毒素或凝集素的神经肽和片段作为神经毒素或 药理拮抗剂研究含神经肽受体的拮抗剂 中枢神经系统（CNS）中的细胞，重点是基础 神经节。 甲状腺激素释放激素（TRH），物质P和生长抑素 将被研究。 使用这些复合物的理由是 毒素毒素或特异性的凝集素的无毒片段 神经肽受体受体细胞通过将毒素络合到A 神经肽。 该项目旨在确定 这种杂种对特异性的基底神经节中的CNS神经元有毒 体内和体外神经肽受体。 使用抗体定向 反对白喉毒素片段，该项目也将尝试 通过免疫组织化学可视化神经元上的神经肽受体。 先前的工作证明了TRH - 二皮细胞菌的受体特异性毒性 毒素杂种在体外与大鼠垂体肿瘤细胞。 这个项目将 建立脊柱中TRH毒素杂种体内毒性的条件 绳索和垂体，存在TRH纤维和受体的区域， 检查全身后的形态变化和受体可视化 或术后交付复合物。 建立了可行性 CNS神经元，神经肽物质P和 生长抑素，与毒素复合，将评估受体结合 和介脑，纹状体或原发性培养物的体外毒性 皮质神经元。 物质P和生长抑素毒素的体内分析 复合物将通过本地在nigrostriatal系统中进行 脑内注射和随后的骨膜变化 神经递质的代谢和行为取决于完整性 黑质体系。 该工具可以提供一种更好地理解流程的方法 负责某些CNS神经元对明显的选择性脆弱性 神经系统的退行性疾病过早死亡，尤其是 在基底神经节（例如：帕金森氏症，亨廷顿，阿尔茨海默氏症）中 允许（1）神经肽的功能和药理分析 神经元，（2）表征神经肽受体在 验尸组织，（3）分析调节的因素 对肽毒素杂种的退化反应，以及（4）选择性模型 神经元丧失。