OPIOD EFFECTS ON SUBSTANTIA NIGRA AND OTHER NEURONS

各种内源性神经递质对神经元功能的调节 阿片肽是最重要的细胞事件之一， 大脑功能 然而，对细胞和细胞外基质的了解并不多， 阿片作用的膜机制。 的目的 一项拨款建议是阐明离子通道， 两种内源性阿片肽的信使机制， 内啡肽和强啡肽 两个脑的原代培养物 出生后动物的区域，蓝斑和 将使用黑质。 电生理技术 包括全细胞形式的膜片钳和细胞内 将使用微电极方法。 该提案分为 两个项目。 第一个项目涉及β-内啡肽对 分离培养的去甲肾上腺素能神经元 蓝核 实验室的初步研究表明，β- 内啡肽在静止凝视时产生缓慢的超极化， 导致神经元活性的抑制。 初步 研究还表明，百日咳毒素抑制了这一点， 超极化，表明抑制性GTP结合蛋白 可能是第二信使 进一步调查所 精确的离子和第二信使机制， 超极化是第一个项目的目标。 第二个项目包括两个部分。 第一部分是 进行黑质原代分离细胞培养， 产后大鼠和小鼠。 多巴胺能神经元将被识别 用免疫组织化学或组织荧光法 方法. 第二部分是对我国农村社会保障制度的影响进行考察。 强啡肽对培养的黑质神经元的作用， 已知富含含强啡肽神经 纤维 强啡肽的离子和第二信使机制 将研究影响。

The modulation of neuronal function by various kinds of endogenous opioid peptides is one of the most important cellular events of brain function. However, not much is known about the cellular and membranous mechanisms of opioid actions. The objective of this grant proposal is to elucidate the ionic channel and second messenger mechanisms of two kinds of endogenous opioid peptides, beta-endorphin and dynorphin. Primary cultures from two brain regions of postnatal animals, the locus coeruleus and the substantia nigra will be used. Electrophysiological techniques including the whole cell version of patch clamp and intracellular microelectrode methods will be used. The proposal is divided into two projects. The first project deals with the effects of beta-endorphin on dissociated cultured noradrenergic neurons from the locus coeruleus. A preliminary study in the laboratory shows that beta- endorphin produces a slow hyperpolarization at the resting stare, resulting in an inhibition of neuronal activity. The preliminary study also shows that pertussis toxin inhibits this hyperpolarization, suggesting that inhibitory GTP-binding proteins may be involved as second messengers. To investigate further the precise ionic and second messenger mechanisms of this slow hyperpolarization is the aim of the first project. The second project consists of two parts. The first part is to make primary dissociated cell culture of the substantia nigra of postnatal rats and mice. Dopaminergic neurons will be identified by using immunohistochemical or cathecholamine histofluorescence methods. The second part is to investigate the effects of dynorphin on cultured neurons from the substantia nigra, which is known to be richly innervated with dynorphin-containing nerve fibers. The ionic and second messenger mechanisms of dynorphin effects will be investigated.