REGULATION OF PITUITARY OPIATE PEPTIDE SECRETION

本申请概述了对内在神经调节机制的研究。 产生内源性阿片剂和相关多肽的细胞。少校 需要检验的假设是特定的抑制和刺激 存在调节阿片黑素皮质素多肽分泌的机制。至 检验多肽是抑制还是刺激的假说 分泌直接发生在阿片黑素皮质素细胞，特异性 神经递质和神经毒素、麻醉性拮抗剂药物和选定的 阿片剂或下丘脑多肽将用于啮齿动物脑垂体腺 体外培养。因为抑制或刺激多肽分泌也可以 受中枢神经系统机制影响，体内完整的啮齿动物 动物模型，将用同样的神经毒素或麻醉剂治疗 以拮抗剂为体外研究对象。衡量…的产出水平 来自体外或体内系统的多肽，生物测定或 黑素细胞刺激素放射免疫测定法 对于β-内啡肽，将使用。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 脑下垂体和脑内神经递质。脑垂体放射免疫测定法 将对组织中的β-内啡肽水平进行比较 多肽的储存水平和血清中释放的物质。最后， 阿片黑素皮质素细胞的细胞学和免疫组织化学研究 将采用定性和定量的方法进行评估 在与化验多肽相同的试验组内， 确定单独的脑下垂体分泌细胞群对 抑制或刺激。预计神经毒剂或 阿片类拮抗剂药物可能对刺激或 预防脑垂体肽的分泌，取决于药剂的 对特定神经递质的选择性。体外和体内试验的应用 对啮齿动物脑垂体腺模型的活体研究，将为 影响内源性阿片剂释放和储存的具体机制 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.