IMMUNOHISTOCHEMICAL MAPPING OF ENKEPHALIN NEURONS

神经肽是近年来从神经组织中分离出来的一类物质 并被发现影响神经系统的活动， 细胞膜、神经元和有行为的有机体。 的免疫组织化学 这些物质揭示了离散的和相当广泛的神经肽 神经元胞体、轴突和终末免疫反应性。 这项建议 重点是确定两个差异分布的细节， 这些肽--蛋氨酸-和亮氨酸-脑啡肽。 除了有 我们的初步研究揭示了脑啡肽能系统的广泛性， 脑啡肽在其拟吗啡性质方面是最重要的， 阿片受体以及器官水平。 这项建议旨在 通过纯化抗血清测定脑啡肽的差异分布， 通过去除脑啡肽上的交叉反应抗体， 异源免疫吸附剂。 纯化的单特异性抗血清， 脑啡肽将用于免疫荧光组织化学程序， 大鼠脑切片，以显示终末、轴突和胞体 含有甲硫氨酸-或亮氨酸-脑啡肽免疫反应性。 的 这些研究的结果将被转化为详细的，示意性的冠状 具有近似立体定位坐标的大鼠大脑的地图。 的 将研究选定的核周体和终端的连通性， 脑立体定向毁损后大鼠脑组织免疫荧光组织化学研究 脑啡肽能胞体。 正常荧光端子的消失将 指示这些终端的起源的胞体。 选定区域将 也可以用其他神经肽的抗血清进行研究， 已经报道了脑啡肽能和其它肽能系统。 最后 该提案的一小部分（10%）旨在调查 脑啡肽在脑内选定核团中的差异分布 使小鼠耐受并依赖阿片类药物。

The neuropeptides are a class of substances recently isolated from neural tissue and have been found to influence the activity of the nervous system at the level of the membrane, the neuron and the behaving organism. Immunohistochemistry of these substances has revealed discrete and rather extensive neuropeptide immunoreactivity in neuronal perikarya, axons and terminals. This proposal focuses on determining the details of the differential distribution of two of these peptides --methionine- and leucine-enkephalin. In addition to the extensiveness of enkephalinergic systems revealed in our preliminary studies, the enkephalins are of interist in light of their morphinomimetic properties at opiate receptors as well as on an organismic level. This proposal seeks to determine the differential distribution of enkephalins by purifying antisera to mono-specificity for each enkephalin by removing cross-reacting antibodies on a heterologous immunoabsorbent. The purified, monospecific antisera to each enkephalin will then be used in immunofluorescence histochemical procedures on sections of the rat brain to reveal the terminals, axons and perikarya containing either methionine- or leucine-enkephalin immunoreactivity. The results of these studies will be translated into detailed, schematic coronal maps of the rat brain with approximate stereotaxic coordinates. The connectivity of selected perikarya and terminals will be studied with immunofluorescence histochemistry on rat brains after stereotaxic lesions of enkephalinergic perikarya. Disappearance of normally-fluorescent terminals will indicate the perikarya of origin for these terminals. Selected regions will also be studied with antisera to other neuropeptides where convergence of enkephalinergic and other peptidergic systems has been reported. Finally, a small portion of this proposal (10%) aims to investigate possible changes in the differential distribution of the enkephalins in selected nuclei in the brains of mice made tolerant to and dependent upon opiates.