ULTRASTRUCTURE AND CONTROL OF ENDORPHIN CELLS

该申请概述了对内源性神经调节的研究 产生内源性阿片样物质的细胞的机制和相关的 缩氨酸 要检验的主要假设是， 存在抑制和刺激机制，其调节 阿黑皮素肽的分泌。 为了验证这个假设 是抑制还是刺激肽分泌 直接作用于阿黑皮素细胞，特异性神经递质， 神经递质、麻醉拮抗剂药物和选择的阿片类药物或 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可能 也受到中枢神经系统机制的影响， 体内啮齿类动物模型将用相同的 神经毒素或麻醉剂拮抗剂作为体外研究。 到 测量来自体外细胞的肽的输出水平， 或用于黑素细胞的体内系统、生物测定或放射免疫测定， 刺激激素或β-内啡肽放射免疫测定法， 被利用 神经毒素在体内的具体作用将得到证实 通过高压液相色谱（HPLC）分析， 垂体和脑神经递质。 的放射免疫测定 垂体组织β-内啡肽水平将在 为了比较肽的储存水平与释放的材料 在血清中。 最后，细胞学和免疫组织化学研究， 阿黑皮素细胞，使用定性和定量方法 为了进行评价，将在相同的实验中进行 组作为那些测定肽，以确定如何个人 垂体分泌细胞群对抑制或 刺激. 预计神经毒剂或阿片类药物 拮抗剂药物可能对刺激有直接作用， 预防肽分泌，这取决于药剂的 对特定神经递质的选择性。 使用体外 在啮齿动物脑垂体模型的体内研究将 为影响释放的特定机制提供支持， 内源性阿片分子的储存。

The 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 neurotoxions, narcotic antagonist drugs, and selected opiate or hypothalamic peptides, will be used in rodent pituitary glands in vitro. Since inhibition or stimulation of peptides secretion may also be affected by central nervous system mechanisms, and 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 radio-immunoassay 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 peptide secretion, depending upon the agent's 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.