ORGANIZATION OF NOCICIPTION DRIVEN NEURONS

伤害驱动神经元的组织

• 批准号: 2265921
• 负责人: A CLAUDIO CUELLO
• 金额: $ 15.16万
• 依托单位: MCGILL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2265921
• 关键词: afferent nerve; antigens; calcitonin gene related peptide; cats; dendrites; dorsal horn; electron microscopy; electrostimulus; enkephalins; horseradish peroxidase; immunocytochemistry; light microscopy; monoclonal antibody; neural information processing; neuroanatomy; neurophysiology; nontherapeutic iontophoresis; pain; spinal nerves; substance P; synapses

The research outlined in this application was devised with the specific aim of clarifying whether the spinal modulation of the nociceptive sensory information, transmitted by peptidergic sensory fibers, occurs at a pre- or postsynaptic site. Our hypothesis, based on the available evidence, is that both substance P- containing sensory fibers and encephalonergic spinal neurons act postsynaptically on glomerular or non-glomerular dendrite's of noniception driven dorsal horn neurons. For that purpose, a combination of ultrastructural immunocytochemistry with intracullular recordings and injections of nociception driven neurons will be used. Such an approach is the only one that can provides a direct answer to the question outlined above, and has never been used before in a systematic way. All the experiments will be carried out on adult cats under alpha-chloralose anesthesia. Nociception driven neurons of the lumour spinal cord will be injected intracellularly with horseradish peroxidense (HRP). The animals will bs fixed by vascular perfusion and the relevant segments of the spinal cord sectioned in a vibratome and processed for the demonstration of peroxidase. The slices containing intracellularly injected cells will be further processed for the demonstration of substance P or encephalon immoractivities at the electron microscoPic level. The antibodies to be used are the P4C1 (anti-substance P/anti-HRP bi-specific antibody) and the NOC1 (antienkeplalin monoclonal antibody). The sensory origin of the substance P fibers will be astablished in some animals by injections of tritiated amino acids in the dorsal root ganglia, and combining radioautography with immunocytochemistry and electrophysiology. At a second stage, a triple labeling involving intracellular injection and simultaneous immunostaining for both peptides in the same preparation will be attempted. For that, diaminobenzidine-based immunocytochemistry will be combined either with radioimmuocytochemistry (using the internally labeled version of the NOC1 antibodys or immunogold. The detection of immunoreactivities for GABA, dynorphin, serotonin, choline acetyltransferase and somatostatin in combination with electrophysiology will be also carried out. The research outlined above is expected to bring important new information on the modulation of pain in the spinal cord.

该应用程序中概述的研究是由 阐明是否的脊柱调节的具体目的是 伤害性感觉信息，通过肽吉尼的感觉传递 纤维，发生在突触前或突触后部位。 我们的假设， 基于可用的证据，是两种物质p- 含有感觉纤维和脑能神经元法 在肾小球或非胶质性树突上的突触 非记录驱动的背角神经元。 为此， 超微结构免疫细胞化学与 胸部记录和伤害感受驱动的注射 神经元将被使用。 这样的方法是唯一可以 直接回答了上面概述的问题，并有 以前从未以系统的方式使用。 所有实验 将在α-氯洛糖的成年猫上进行 麻醉。 腔脊髓的伤害感受驱动的神经元 将在细胞内注射辣根过氧化物 （HRP）。 动物将通过血管灌注固定和 脊髓的相关段，分为振动型和 处理过过氧化物酶的演示。 切片 包含细胞内注射的细胞将进一步处理 为了证明物质P或脑不活活 在电子显微镜水平上。 要使用的抗体是 P4C1（抗固定P/抗HRP BI特异性抗体）和 NOC1（抗杀菌蛋白单克隆抗体）。 感官起源 物质P纤维将通过某些动物的特征 在背根神经节中注射tri的氨基酸， 并将放射学与免疫细胞化学和 电生理学。 在第二阶段，涉及三重标签 两者的细胞内注射和同时进行免疫染色 将尝试同一制剂中的肽。 为此， 将组合基于二氨基苯胺的免疫细胞化学化学 使用放射量摄影化学（使用内部标记的版本 NOC1抗体或免疫金的。 检测 GABA，Dynorphin，5-羟色胺，胆碱的免疫反应性 乙酰转移酶和生长抑素与 电生理学也将进行。 研究概述了 上面有望带来有关的重要新信息 调节脊髓疼痛。