REGULATION OF PITUITARY OPIATE PEPTIDE SECRETION

本申请概述了对内在神经调节机制的研究 产生内源性鸦片和相关肽的细胞。 主要 要检验的假设是，特异性抑制和刺激 存在调节阿黑皮素肽分泌的机制。 到 检验是否抑制或刺激肽的假说 分泌直接发生在阿黑皮素细胞上，特异性 神经递质和神经毒素，麻醉拮抗剂药物， 阿片或下丘脑肽将用于啮齿动物脑垂体， 体外 由于肽分泌的抑制或刺激也可能是 受中枢神经系统机制的影响，一个完整的体内啮齿动物 动物模型，将用相同的神经毒素或麻醉剂处理 拮抗剂作为体外研究。 衡量产出水平 来自体外或体内系统的肽，生物测定或 促黑素细胞激素放射免疫测定法或放射免疫测定法 β-内啡肽，将被使用。 神经毒素在体内的特异性作用 将通过高压液相色谱（HPLC）分析进行确认 脑垂体和脑神经递质的作用 垂体放射免疫测定法 将进行β-内啡肽的组织水平测定， 血清中肽与释放物质的储存水平。 最后， 阿黑皮素细胞的细胞学和免疫组织化学研究，使用 定性和定量的评价方法，将进行 在与测定肽的实验组相同的实验组中， 确定各组垂体分泌细胞如何响应 抑制或刺激。 预计神经毒剂或 阿片拮抗剂药物可能对刺激有直接作用， 预防垂体肽分泌，具体取决于药物的 对特定神经递质的选择性。 在体外和体内使用 啮齿动物脑垂体模型的体内研究，将为 影响内源性阿片样物质释放和储存的特殊机制 分子。

This application outlines a study on intrinsic neuroregulatory mechanisms of cells which produce endogenous opiate and related peptides. The major hypothesis to be tested is that specific inhibitory and stimulatory mechanisms exist which regulate secretion of opiomelanocortin peptides. To examine the hypothesis of whether inhibition or stimulation of peptide secretion occurs directly on opiomelanocortin cells, specific neurotransmitters and neurotoxins, narcotic antagonist drugs, and selected opiate or hypothalamic peptides will be used in rodent pituitary glands in vitro. Since inhibition or stimulation of peptide secretion may also be affected by central nervous system mechanisms, an intact in vivo rodent animal model, will be treated with the same neurotoxins or narcotic antagonists as the in vitro studies. To measure the level of output of peptides from either the in vitro or in vivo systems, bioassay or radioimmunoassay for melanocyte-stimulating hormone, or radioimmunoassay for beta-endorphin, will be used. Specific effects of neurotoxins in vivo will be confirmed by high pressure liquid chromatographic (HPLC) analysis of pituitary and brain neurotransmitters. Radioimmunoassay for pituitary tissue levels of beta-endorphin will be performed in order to compare stored levels of peptide with released material in serum. Finally, cytologic and immunohistochemical studies of opiomelanocortin cells, using qualitative and quantitative methods for evaluation, will be performed within the same experimental groups as those assayed for peptides, to determine how individual groups of pituitary secretory cells respond to inhibition or stimulation. It is anticipated that neurotoxic agents or opiate antagonist drugs may have direct effects on stimulation or prevention of pituitary peptide secretion, depending upon the agents' selectivity for a particular neurotransmitter. The use of in vitro and in vivo studies on the rodent pituitary gland model, will provide support for specific mechanisms which affect release and storage of endogenous opiate molecules.