Characterizing κ(3) opioid receptors with a selective monoclonal antibody Journal Article
| Authors: | Brooks, A. I.; Standifer, K. M.; Rossi, G. C.; Mathis, J. P.; Pasternak, G. W. |
| Article Title: | Characterizing κ(3) opioid receptors with a selective monoclonal antibody |
| Abstract: | To help characterize κ3 receptors and establish their relationship to traditional μ and δ receptors, we have generated a κ3-selective monoclonal antibody. Monoclonal antibodies were raised against BE(2)-C cells, a human neuroblastoma cell line containing μ, κ3, and δ opioid receptors. Of the 5,000 hybridoma cell lines screened, approximately 2,000 hybridomas tested positive against BE(2)-C membranes by ELISA, but only 98 of these were negative against a different neuroblastoma cell line lacking opioid receptors. Supernatants from one hybridoma, 8D8, inhibited up to 90% of 3H- NalBzoH (κ3) binding without affecting 3H-DAMGO (μ) or 3H-naltrindole (δ) binding in BE(2)-C membranes. The selectivity of the antibody was further demonstrated by its blockade of the inhibition of cAMP accumulation in BE(2)-C cells by the κ3 agonist NalBzoH but not the μ agonist morphine. Monoclonal antibody 8D8 (mAb8D8) also recognizes κ3 receptors from mouse, rat, and calf brain. Administered intracerebroventricularly, mAb8D8 blocked κ3 but not morphine (μ) analgesia in vivo. On Western blots, mAb8D8 recognized a protein with a molecular mass of approximately 70 kilodaltons in BE(2)-C. These studies demonstrate the selectivity of mAb8D8 for κ3 receptors and provide additional support for the existence of this unique opioid receptor subtype. |
| Keywords: | controlled study; nonhuman; brain tumor; brain neoplasms; mouse; animal; metabolism; animals; mice; animal tissue; opiate; drug effect; tumor cells, cultured; enzyme linked immunosorbent assay; monoclonal antibody; mice, inbred strains; blotting, western; immunology; antibodies, monoclonal; narcotic analgesic agent; diagnostic agent; cell culture; neuroblastoma; rat; western blotting; cyclic amp; rats; receptor affinity; mouse strain; enzyme-linked immunosorbent assay; analgesics, opioid; pain measurement; analgesia; pain assessment; receptor binding; binding competition; opioid peptides; binding, competitive; antinociception; kappa opiate receptor; opiate peptide; mu opioid receptor; male; priority journal; article; receptors, opioid, kappa; delta opioid receptor; camp accumulation |
| Journal Title: | Synapse |
| Volume: | 22 |
| Issue: | 3 |
| ISSN: | 0887-4476 |
| Publisher: | John Wiley & Sons |
| Date Published: | 1996-03-01 |
| Start Page: | 247 |
| End Page: | 252 |
| Language: | English |
| DOI: | 10.1002/(sici)1098-2396(199603)22:3<247::aid-syn7>3.0.co;2-c |
| PUBMED: | 9132993 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 22 November 2017 -- Source: Scopus |
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