The κ-Opioid Receptor is Primarily Postsynaptic: Combined Immunohistochemical Localization of the Receptor and Endogenous Opioids

• Published in: Proceedings of the National Academy of Sciences - PNAS 1995-05, Vol.92 (11), p.5062-5066
• Main Authors: Arvidsson, Ulf, Riedl, Maureen, Chakrabarti, Sumita, Vulchanova, Lucy, Lee, Jang-Hern, Nakano, Albert H., Lin, Xiaoqin, Loh, Horace H., Law, Ping-Yee, Wessendorf, Martin W., Elde, Robert
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antibodies; Antibody Specificity; Basal ganglia; Biology; Blotting, Western; Brain; Brain - cytology; Brain - metabolism; Cell Line; Cercopithecus aethiops; Complementary DNA; COS cells; Dynorphins - analysis; Epitopes - analysis; Gene Expression; Guinea Pigs; Immunohistochemistry; Kidney; Ligands; Male; Messenger RNA; Microscopy, Confocal; Molecular Sequence Data; Neuroblastoma; Neurology; Neurons; Neurons - cytology; Neurons - metabolism; Opioid receptors; Organ Specificity; Peptide Fragments - chemistry; Peptide Fragments - immunology; Peptides; Protein Precursors - analysis; Rabbits - immunology; Rats; Rats, Sprague-Dawley; Receptors; Receptors, Opioid, kappa - analysis; Receptors, Opioid, kappa - biosynthesis; Receptors, Opioid, mu - analysis; Recombinant Proteins - analysis; Recombinant Proteins - biosynthesis; RNA, Messenger - analysis; Rodents; Spinal cord; Spinal Cord - cytology; Spinal Cord - metabolism; Transfection
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.11.5062

Antisera were raised against a synthetic peptide 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 epitopetagged 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.