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SYNAPTIC FUNCTIONS IN CULTURE: PEPTIDERGIC AND OTHERS

培养中的突触功能：肽及其他

基本信息

批准号：
3210313
负责人：
DAVID D POTTER
金额：
$ 21.87万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-04-01 至 1993-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3210313
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

项目摘要

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.

该提议源于一项为期 9 年的关于发射机选择的研究培养大鼠交感主要神经元。开发时在在某些培养条件下，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）。为了研究这个神经元发育过程中的转变，我们设计了一种微培养方法允许电生理学特征的发射器状态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在转变时，神经元表现出肾上腺素能：胆碱能双重功能，并且可以通过重复记录来直接进行转换相同的神经元持续长达 45 天。该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。与成人神经节分离的神经元有时也表现出肾上腺素能：胆碱能双重功能。初级感官的平行研究神经元和肌间神经元现在正在顺利进行。近来，几实验室报告称，交感神经元、感觉神经元和肌间神经元的成年大鼠对许多活性肽（5 英寸）表现出免疫反应性仅交感神经元）；有证据表明有几种分泌物对于胺：肽双重功能。这表明发射机选择在这些类型的神经元中，其控制要复杂得多比之前想象的要多。由于上述培养方法允许直接研究突触功能，包括多发射器状态，这在体内通常很困难，我们有开始研究我们文化中的肽能功能。我们有 P 类物质和禽类胰腺的初步证据多肽样免疫反应性。在本提案中，我们请求支持对我们培养的肽能功能的系统研究周围神经元。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）报道的肽体内。 ii) 将神经元与对神经元敏感的靶细胞共培养肽，在新型微培养物中，并关联肽能功能，单个神经元的精细结构和免疫反应性。 iii) 调查正如我们之前所做的那样，控制肽能功能的表达通过肾上腺素能：胆碱能功能完成。长期目标是扩大对发射机选择的理解，这是发射机选择的一个基本方面神经元细胞生物学。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Synaptic functions in rat sympathetic neurons in microcultures. IV. Nonadrenergic excitation of cardiac myocytes and the variety of multiple-transmitter states.

微培养物中大鼠交感神经元的突触功能。

DOI：
10.1523/jneurosci.07-02-00380.1987
发表时间：
1987
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
0
作者：
Matsumoto,SG;Sah,D;Potter,DD;Furshpan,EJ
通讯作者：
Furshpan,EJ

Neurons dissociated from rat myenteric plexus retain differentiated properties when grown in cell culture. III. Synaptic interactions and modulatory effects of neurotransmitter candidates.

从大鼠肌间神经丛分离的神经元在细胞培养物中生长时保留分化的特性。