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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >The κ-opioid receptor is primarily postsynaptic: Combined immunohistochemical localization of the receptor and endogenous opioids
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The κ-opioid receptor is primarily postsynaptic: Combined immunohistochemical localization of the receptor and endogenous opioids

机译：κ阿片受体主要在突触后：受体与内源性阿片类药物的免疫组织化学结合定位

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摘要

Antisera were raised against a synthetic pep-tide corresponding to the carboxyl terminus of the κ-opioid receptor (KOR1). Specificity of the antisera was verified by staining of COS-7 cells transfected with KOR1 and epitope-tagged KOR1 cDNAs, by recognition by the antisera of proteins on Western blots of both transfected cells and brain tissue, by the absence of staining of both brain tissue and transfected cells after preabsorption of the antisera with the cognate peptide, and on the strong correlation between the distribution of KOR1 immunoreactivity and that of earlier ligand binding and in situ hybridization studies. Results indicate that KOR1 in neurons is targeted into both the axonal and somatodendritic compartments, but the majority of im-munostaining was seen in the somatodendritic compartment. In sections from rat and guinea pig brain, prominent KOR1 staining was seen in the ventral forebrain, hypothalamus, thalamus, posterior pituitary, and midbrain. While the staining pattern was similar in both species, distinct differences were also observed. The distribution of preprodynorphin and KOR1 immunoreactivity was complementary in many brain regions, suggesting that KOR1 is poised to mediate the physiological actions of dynorphin. However, the distribution of KOR1 and enkephalin immunoreactivity was complementary in some regions as well. These results suggest that the KOR1 protein is primarily, but not exclusively, deployed to postsynaptic membranes where it mediates the effects of products of preprodynorphin and possibly preproenkephalin.
机译：产生抗血清，以抵抗对应于κ阿片受体（KOR1）羧基末端的合成肽。通过对转染了KOR1和表位标记的KOR1 cDNA的COS-7细胞进行染色，通过对转染的细胞和脑组织的Western印迹上的蛋白质进行抗血清识别，并且对两个脑组织都没有染色，来验证抗血清的特异性关联肽预吸收抗血清后转染的细胞和转染的细胞，以及KOR1免疫反应性的分布与早期配体结合和原位杂交研究之间的强相关性。结果表明，神经元中的KOR1既被定位到了轴突区，也被定位到了树突状区，但是大多数免疫染色都在树突状区中。在大鼠和豚鼠脑的切片中，在前脑腹侧，下丘脑，丘脑，垂体后叶和中脑中可见明显的KOR1染色。尽管两个物种的染色模式相似，但也观察到了明显的差异。前强啡肽和KOR1免疫反应性的分布在许多大脑区域是互补的，这表明KOR1有望介导强啡肽的生理作用。但是，在某些区域，KOR1和脑啡肽的免疫反应性分布也是互补的。这些结果表明，KOR1蛋白主要但非排他地部署在突触后膜上，在该膜上介导前强啡肽和可能的前脑啡肽产物的作用。

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