MOR1--MU OPIOID RECEPTORS AND THEIR ENDOGENOUS LIGANDS

每种药理鉴定的阿片类药物的克隆 受体(即Delta、Mu和kappa)极大地改变了大自然。 在不匹配的问题中可以解决的问题 阿片受体及其内源性配体之间的相互作用。两个问题 问题是：克隆的受体是否解释了所有的阿片类药物？ 神经系统中的受体？“和”已识别的内源性 配体代表可用于作用的全套配体 阿片受体？“ 随着Mu阿片类药物基因敲除小鼠的成功生产和 一个新的神经肽家族的分离和鉴定 内吗啡)，这些问题可能是 以新奇的方式处理。正是考虑到这一点，我们建议 将有助于实现以下目标的实验 具体目标： 1)确定克隆的Mu受体mor1是否是Mu受体 负责突触前抑制去甲肾上腺素的释放 蓝斑神经元的轴突。 2)确定负责合成的生物合成前体 内吗啡，以及表达内吗啡肽的细胞的区域分布 内吗啡素及其生物活性产物的转录本。 3)确定内吗啡肽和内吗啡肽之间的空间关系 MOR1。 4)确定缺乏内吗啡的小鼠的内吗啡素的表达是否发生了变化 表观同源受体(MOR1)。 这些目标将通过几种不同类型的实验来实现。 野生型和MU基因敲除的前脑突触小体制备 小鼠将有助于阐明MOR1在多大程度上负责 去甲肾上腺素释放的突触前抑制。此外，建议的还有 分离生物合成前体蛋白的表达克隆实验 内吗啡素，然后进行原位杂交研究。最后，我们 建议研制抗内吗啡肽和内吗啡肽的抗血清 随后的野生型和野生型的双色免疫荧光实验 确定基因敲除小鼠与克隆小鼠之间的匹配程度 受体和这一新的内源性配体家族。

The cloning of each of the pharmacologically identified types of opioid receptors (i.e., delta, mu and kappa) has dramatically altered the nature of the questions that can be addressed in the problem of the mismatch between opioid receptors and their endogenous ligands. Two questions that arise are: "Do the cloned receptors account for all of the opioid receptors in the nervous system?" and "Do the identified endogenous ligands represent the complete set of ligands available for action at opioid receptors?" With the successful production of a mu opioid knockout mouse and the isolation and characterization of a new family of neuropeptides (the endomorphins) that are active in mu receptors, these questions can be addressed in novel ways. It is with this in mind that we propose experiments that will contribute to the realization of the following specific aims: 1) Determine if the cloned mu receptor, MOR1, is the mu receptor responsible for presynaptic inhibition of norepinephrine release from axons of locus coeruleus neurons. 2) Determine the biosynthetic precursor responsible for synthesis of endomorphin, and the regional distribution of cells that express the transcript for endomorphin and its biologically active products. 3) Determine the spatial relationship between the endomorphin peptides and MOR1. 4) Determine if the expression of endomorphin is altered in mice that lack the apparent cognate receptor (MOR1). These aims will be accomplished by several different types of experiments. Synaptosomal preparations of the forebrain of wild type and mu knockout mice will help to elucidate the extend to which MOR1 is responsible for the presynaptic inhibition of norepinephrine release. Also, proposed are expression cloning experiments to isolate the biosynthetic precursor of the endomorphins, followed by in situ hybridization studies. Finally, we propose the development of specific antisera to the endomorphins and subsequent two-color immunofluorescent experiments in wild type and knockout mice to determine the extent of match between the cloned mu receptor and this new family of endogenous ligands.