Chronic opioid treatment attenuates carbachol-mediated polyphosphoinositide hydrolysis in chick embryo neuronal cultures.
Opiate binding sites on cultured neurons derived from 6-day-old (E6) chick embryo cerebral hemispheres (CH), shown to be cholinergic by choline acetyltransferase immunostaining, were labeled with [3H]etorphine (mu and delta opiate receptors expression) and [3H]morphine (mostly mu). When examined by light microscope autoradiography, opiate receptors were found to be expressed by most neurons, and were distributed predominantly on neuronal perikarya. Muscarinic and opiate receptors in E6CH cultured neurons were found to be functionally coupled when the effects of opiate receptor occupancy on the inositol phosphate-linked muscarinic receptors was studied. Carbachol stimulated the release of [3H]inositol phosphates (InsP) from cultures preincubated with [3H]inositol and LiCl, in a dose-dependent manner, and the functional expression of muscarinic receptors peaked in number at day 7 in culture, declining thereafter. Short-term (less than 1 h) treatment of E6 neuronal cultures with 1 microM opioid peptides such as morphiceptin or D-Ala2-D-Leu5-enkephalin (DADLE) did not inhibit the release of inositol phosphates in response to 1 mM carbachol whereas forskolin, which also activates adenylate cyclase and raises cAMP levels, inhibited InsP release by about 25%. In contrast, long-term (48 h) opioid treatment with either morphiceptin or DADLE (1-10 microM) inhibited the carbachol-stimulated inositol phosphate release by greater than or equal to 50%. Prolonged treatment with morphiceptin also inhibited the bradykinin-mediated release of InsP from E6CH cells. In both cases, the inhibition was partially blocked by the continuous presence of naloxone, suggesting that the inhibition was mediated through opiate receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
Mangoura
D
D
Department of Pediatrics, University of Chicago Medical School, IL 60637.
Dawson
G
G
D000700:Analgesics / Q000494:pharmacology*; D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D001921:Brain / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo; D004723:Endorphins / Q000494:pharmacology*; D016308:Enkephalin, Leucine-2-Alanine / Q000494:pharmacology*; D006868:Hydrolysis; D007294:Inositol / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism; D007700:Kinetics; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000378:metabolism*; D011957:Receptors, Opioid / Q000187:drug effects / Q000378:metabolism*
D000700:Analgesics; D004723:Endorphins; D007295:Inositol Phosphates; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D011957:Receptors, Opioid; D007294:Inositol; D016308:Enkephalin, Leucine-2-Alanine; D002217:Carbachol; C028889:morphiceptin; D001920:Bradykinin