SYNAPTIC FUNCTIONS IN CULTURE: PEPTIDERGIC AND OTHERS

培养中的突触功能：肽及其他

• 批准号: 3210314
• 负责人: DAVID D POTTER
• 金额: $ 16.27万
• 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/3210314
• 关键词: afferent nerve; histochemistry /cytochemistry; immunochemistry; neural plasticity; neurons; 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.

这项建议是从一项长达9年的关于发射机选择的研究中提出的， 培养的大鼠交感主神经元。 当开发在 在一定的培养条件下，这些神经元可以从肾上腺素能神经元转移到肾上腺素能神经元。 胆碱能状态（神经元可塑性的新形式）。 研究这个 我们设计了一种微培养方法， 允许电生理学表征的 单个神经元与其精细结构直接相关， 末端或其免疫反应性。 通过这种方法，我们发现， 过渡，神经元显示肾上腺素能：胆碱能双重功能， 过渡可以直接通过重复记录从 相同的神经元在45天内的变化。 该方法还显示， 一些神经元分泌另一种递质，可能是腺苷， 胺。 从成人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的研究进展顺利。 最近有数 实验室已经报道了交感神经、感觉神经和肌间神经元 成年大鼠对许多活性肽（5 in 交感神经元单独）;有证据表明，分泌的几个， 用于胺：肽双功能。 这表明发射机的选择 在这些类型的神经元中， 比以前想象的。 由于上述培养方法 允许直接研究突触功能，包括 多递质状态，这在体内往往是困难的，我们有 开始在我们的培养物中研究肽能功能。 我们有 初步证据表明，P物质和禽胰腺 多肽样免疫反应性。 在本提案中，我们请求支持 系统地研究了我们培养的细胞的肽能功能， 外周神经元 我们的具体目标是：（一）与传统的 免疫细胞化学方法筛选四种类型的培养神经元 （交感神经、感觉神经、肌间神经和副交感神经） in vivo. ii）将神经元与对所述神经元敏感的靶细胞共培养。 肽，在新的微培养物，并相关肽能功能， 单个神经元的精细结构和免疫反应性。 ㈢调查 如我们先前所述， 肾上腺素能和胆碱能功能。 长期目标是 扩大对变送器选择的理解，这是 神经细胞生物学