Arvidsson U, Riedl M, Chakrabarti S, Vulchanova L, Lee J H, Nakano A H, Lin X, Loh H H, Law P Y, Wessendorf M W
Department of Cell Biology, University of Minnesota, Minneapolis 55455, USA.
Proc Natl Acad Sci U S A. 1995 May 23;92(11):5062-6. doi: 10.1073/pnas.92.11.5062.
Antisera were raised against a synthetic peptide corresponding to the carboxyl terminus of the kappa-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 immunostaining 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.
制备了针对与κ-阿片受体（KOR1）羧基末端对应的合成肽的抗血清。通过对转染了KOR1和表位标记的KOR1 cDNA的COS-7细胞进行染色、通过抗血清对转染细胞和脑组织的蛋白质印迹上的蛋白质的识别、通过用同源肽预吸收抗血清后脑组织和转染细胞均无染色以及通过KOR1免疫反应性分布与早期配体结合和原位杂交研究结果之间的强相关性，验证了抗血清的特异性。结果表明，神经元中的KOR1定位于轴突和树突-胞体区室，但大部分免疫染色见于树突-胞体区室。在大鼠和豚鼠脑切片中，在前脑腹侧、下丘脑、丘脑、垂体后叶和中脑可见明显的KOR1染色。虽然两种物种的染色模式相似，但也观察到了明显差异。在许多脑区，前强啡肽原和KOR1免疫反应性的分布是互补的，这表明KOR1可能介导强啡肽的生理作用。然而，在某些区域，KOR1和脑啡肽免疫反应性的分布也是互补的。这些结果表明，KOR1蛋白主要但并非唯一地定位于突触后膜，在那里它介导前强啡肽原产物以及可能的前脑啡肽原产物的作用。
