ULTRASTRUCTURE AND CONTROL OF ENDORPHIN CELLS

该申请概述了一项关于内在神经调节的研究 细胞产生内源性阿片及其相关物质的机制 多肽。需要检验的主要假设是， 存在抑制和刺激机制，这些机制调节 阿片黑素皮质肽的分泌。来检验这一假设 是否会抑制或刺激多肽分泌 直接作用于阿片黑素皮质素细胞、特定的神经递质和 神经毒素、麻醉性拮抗剂药物和选定的阿片类药物或 下丘脑肽，将用于啮齿动物脑下垂体腺 体外培养。因为抑制或刺激多肽分泌可能 也受到中枢神经系统机制的影响，并且完好无损 在活体啮齿动物模型中将用同样的方法处理 神经毒素或麻醉性拮抗剂作为体外研究。至 测量来自体外的任何一个多肽的产量水平 或体内系统，黑素细胞的生物测定或放射免疫测定- 刺激素，或β-内啡肽的放射免疫分析，将 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 脑下垂体和脑内神经递质。放射免疫测定法 脑垂体组织中的β-内啡肽水平将在 为了比较多肽的储存水平和释放的材料 在血清中。最后，细胞学和免疫组织化学研究 阿片黑素皮质素细胞，使用定性和定量方法 为了进行评估，将在相同的实验中进行 组作为那些检测多肽的组，以确定个体如何 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或 预防多肽分泌，取决于试剂的 对特定神经递质的选择性。试管苗的应用 而对啮齿动物脑垂体腺模型的体内研究将 为影响发布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.