SYNAPTIC FUNCTIONS IN CULTURE: PEPTIDERGIC AND OTHERS

培养中的突触功能：肽及其他

• 批准号: 3210311
• 负责人: DAVID D POTTER
• 金额: $ 17.08万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3210311
• 关键词: afferent nerve; histochemistry /cytochemistry; immunochemistry; laboratory rat; neural plasticity; neurons; neuropeptides; parasympathetic nervous system; peripheral nervous system; sympathetic nervous system; synapses; tissue /cell culture

This proposal has arisen from a 9-year old study of transmitter choice in cultured sympathetic principal neurons of the rat. When developing in certin culture conditions, these neurons can be shifted from adrenergic to cholinergic status (a new form of neuronal plasticity). To study this transition in developing neurons, we devised a microculture method which permits the electrophysiologically-characterized transmitter status of a single neuron to be correlated directly with the fine structure of its endings or its immunoreactivity. With this method we found that during the transition, the neurons display adrenergic:cholinergic dual function, and that the transition can be followed directly by recording repeatedly from the same neuron over periods up to 45d. This method also revealed that some neurons secreted another transmitter, probably adenosine, with one of the amines. Neurons dissociated from adult ganglia also sometimes display adrenergic:cholinergic dual function. Parallel studies of primary sensory neurons and myenteric neurons are now well underway. Recently, several laboratories have reported tha sympathetic, sensory, and myenteric neurons of adult rats disply immunoreactivity for many active peptides (5 in sympathetic neurons alone); there is evidence for secretion of several, nd for amine:peptide dual function. This indicates that transmitter choice and its control are considerably more complex in these types of neurons than previously supposed. Since the culture methods mentioned above permits a direct study of synaptic functions, including multiple-transmitter status, which is often difficult in vivo, we have begun investigation of peptidergic functions in our cultures. We have preliminary evidence for Substance P-like and Avian Pancreatic Polypeptide-like immunoreactivity. In this proposal we request support for a systematic investigation of peptidergic functions in our cultured peripheral neurons. Our specific aims are: i) With conventional immunocytochemical methods to screen four types of cultured neurons (sympathetic, sensory, myenteric and parasympathetic) for peptides reported in vivo. ii) To co-culture the neurons with target cells sensitive to the peptides, in the novel microcultures, and to correlate peptidergic functon, fine structure and immunoreactivity in single neurons. iii) To investigate the control of expression of peptidergic functions as we have previously done with adrenergic:cholinergic functions. The long-term objective is to enlarge understanding of transmitter choice, a fundamental aspect of neuronal cell biology.

这一建议源于一项关于发射机选择的9年前的研究 培养的大鼠交感神经主神经元。在中开发时 在一定的培养条件下，这些神经元可以从肾上腺素能转变为 胆碱能状态(神经元可塑性的一种新形式)。研究这一点 在发育中的神经元的过渡，我们设计了一种微培养方法， 允许具有电生理特征的发射器状态 单个神经元与其精细结构直接相关 结局或其免疫反应性。通过这种方法，我们发现在 过渡期，神经元表现出肾上腺素能：胆碱能双重功能，以及 在过渡之后可以直接从 相同的神经元持续时间长达45d。这一方法还揭示了 一些神经元分泌另一种递质，可能是腺苷，其中之一 胺类物质。从成年神经节分离出来的神经元有时也会显示 肾上腺素能：胆碱能双重功能。初级感觉的平行研究 神经元和肌间神经元目前进展顺利。最近，有几个 实验室已报道交感神经元、感觉神经元和肌间神经元 的成年大鼠表现出对许多活性多肽的免疫反应(5in 单独的交感神经元)；有证据表明有几个和 对于胺：多肽具有双重功能。这表明发射机的选择 在这些类型的神经元中，其控制要复杂得多 比之前设想的要好。由于上面提到的培养方法 允许直接研究突触功能，包括 多发射器状态，这在体内通常是困难的，我们有 开始研究我们文化中的肽能功能。我们有 类P物质与禽类胰腺的初步证据 多肽样免疫反应。在本提案中，我们请求支持 我们培养的人的肽能功能的系统研究 外周神经元。我们的具体目标是：i)与传统 免疫细胞化学方法筛选四种培养神经元 (交感、感觉、肌间和副交感) 在活体内。Ii)将神经元与敏感的靶细胞共培养 多肽，在新的微培养中，并与多肽能功能相关， 单个神经元的精细结构和免疫反应。三)进行调查 如前所述，对多肽能功能表达的控制 肾上腺素能：胆碱能功能。长期目标是 扩大对发射机选择的理解，这是 神经细胞生物学。