IMMUNOHISTOCHEMICAL MAPPING OF ENKEPHALIN NEURONS

神经肽是最近从神经组织中分离出来的一类物质。 并已被发现影响神经系统的活动 由膜、神经元和行为有机体组成。血管内皮细胞的免疫组织化学 这些物质揭示了离散的和相当广泛的神经肽 神经元核膜、轴突和终末的免疫反应。这项建议 重点是确定两个 这些多肽--蛋氨酸和亮氨酸-脑啡肽。除 在我们的初步研究中揭示了脑啡肽能系统的广泛性， 根据脑啡肽的形态仿射特性，脑啡肽具有内部结构。 阿片受体以及在生物体水平上。这项建议旨在 纯化脑啡肽抗血清以确定脑啡肽的差异性分布 通过去除脑啡肽上的交叉反应抗体对每个脑啡肽进行单特异性检测 异种免疫吸收剂。纯化的、单特异性的抗血清 脑啡肽随后将用于免疫荧光组织化学程序 显示终末、轴突和核周膜的大鼠脑切片 含有蛋氨酸或亮氨酸脑啡肽免疫反应性。这个 这些研究的结果将被翻译成详细的、示意性的冠状 用近似的立体定位坐标绘制的大鼠大脑地图。这个 将研究选定的Perkarya和航站楼的连通性 脑立体定向毁损后大鼠脑的免疫荧光组织化学研究 脑啡肽能周围症。正常荧光终端的消失将 标明这些码头的原产地。选定的地区将 也可以用其他神经肽的抗血清进行研究，其中收敛于 脑啡肽能系统和其他肽能系统已有报道。最后，一个 这项提案的一小部分(10%)旨在调查 脑啡肽在大鼠脑内部分核团中的差异分布 使小鼠对鸦片类药物产生耐受性和依赖性。

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.