REGULATION OF NEUROPEPTIDE BIOSYNTHESIS AND SECRETION

本研究的目的是确定神经元的中枢调节因子， 使用培养的上级颈神经节（SCG）神经元作为模型。 这项建议的重点是研究规管的相互关系 神经肽和经典递质表达之间的关系， 特别是神经肽Y（NPY）和儿茶酚胺（CA）的表达， 在SCG。 考察因素和环境线索的几个问题 协同或独立调节SCG、NPY和儿茶酚胺（CA） 表达将被解决，包括（1）什么是监管机构 神经元NPY表达(2)NPY和CA的表达是否被相同的 监管因素？(3)神经元表型如何影响NPY表达？ (4)特异性靶组织调节神经元NPY表达吗？(5)什么 是改变神经肽表达的细胞机制 几 参数将用于表征培养物NPY/CA含量， 生物合成，包括放射免疫测定，生物合成标记技术， 北方印迹分析和HPLC/电化学检测技术。 的 神经元输入对SCG细胞的影响，即刺激第二 信使系统、钙流、去极化和特异性受体- 将检查对SCG NPY产生的介导刺激并将其与 生产水平。 如何改变SCG神经递质的因素 从儿茶酚胺能表型到胆碱能表型的表达改变NPY 将检查表达。 在共培养研究中，SCG 靶细胞对神经递质/神经肽表达的影响 测定 这些研究将对以下方面产生重大影响： 建立复杂和整合的信号如何引导正常的神经元 发育和生理功能。

The goal of this study is to determine the central regulators of neuronal function using cultured superior cervical ganglion (SCG) neurons as models. The focus of this proposal is to examine the regulatory interrelationship between neuronal peptide and classical transmitter expression, and in particular, the expression of neuropeptide Y (NPY) and catecholamines (CA) in the SCG. Several questions examining the factors and environmental cues that coordinately or independently regulate SCG NPY and catecholamine (CA) expression will be addressed, including (1) What are the regulators of neuronal NPY expression? (2) Are NPY and CA expression altered by the same regulatory factors? (3) How does neuronal phenotype affect NPY expression? (4) Do specific target tissues regulate neuronal NPY expression? (5) What are the cellular mechanisms of altered neuropeptide expression? Several parameters will be used to characterize culture NPY/CA content and biosynthesis including radioimmunoassay, biosynthetic labeling techniques, Northern blot analysis, and HPLC/electrochemical detection techniques. The effects of neuronal input onto SCG cells, that is stimulation of second messenger system, calcium flux, depolarization, and specific receptor- mediated stimulation on SCG NPY production will be examined and correlated to CA production levels. How factors that change SCG neurotransmitter expression from catecholaminergic to cholinergic phenotype alter NPY expression will be examined. In coculture studies, the effects of SCG target cells on neurotransmitter/neuropeptide expression will be determined. These studies will have significant implications in establishing how complex and integrated signals direct normal neuronal development and physiological functions.