Modulation of immunoreactive protein kinase C-alpha and beta isoforms and G proteins by acute and chronic treatments with morphine and other opiate drugs in rat brain.
The abundance of protein kinase C-alpha and beta isoforms (PKC-alphabeta), PKC-alpha messenger (m) RNA and guanine nucleotide-binding G protein subunits (G alpha(i1/2), G alpha(o), and G beta) were quantitated in the rat cerebral cortex after acute and chronic treatments with various opiate drugs. Acute (100 mg/kg for 2 h) and chronic (10 to 100 mg/kg for 5 days) treatment with morphine decreased similarly the immunoreactivity of PKC-alphabeta (28% and 32%, respectively). Acute (2 h) and chronic treatment (5 days) with other mu-agonists heroin (30 mg/kg and 10 to 30 mg/kg) and methadone (30 mg/kg and 5 to 30 mg/kg) also induced similar decreases of PKC-alphabeta (acute: 25% and 23%; chronic: 28% and 18%). After the chronic treatments, spontaneous (48 h) or naloxone (2 mg/kg)-precipitated opiate withdrawal (2 h) resulted in up-regulation of PKC-alphabeta above control levels (30-38%), and in the case of morphine withdrawal in a concomitant marked increase in the expression of PKC-alpha mRNA levels (2.3-fold). Acute (2 h) treatments with pentazocine (80 mg/kg, mixed kappa/delta-agonist and mu-antagonist), spiradoline (30 mg/kg, selective kappa-agonist) and [D-Pen2, D-Pen5] enkephalin (14 nmol i.c.v., selective delta-agonist) induced significant decreases of PKC-alphabeta (19-33%). Chronic (5 days) treatment with pentazocine (10 to 80 mg/kg), but not spiradoline (2 to 30 mg/kg), also induced a similar decrease of PKC-alphabeta (35%). In pentazocine- or spiradoline-dependent rats, naloxone (2 mg/kg) did not induce up-regulation of brain PKC-alphabeta. Acute (10 mg/kg for 2 h) and chronic (2x10 mg/kg for 5 and 14 days) treatment with naloxone did not alter PKC-alphabeta immunoreactivity. Chronic, but not acute, treatment with mu-agonists (morphine, heroin and methadone) increased the immunoreactivities of G alpha(i1/2) (33-37%), G alpha(o), (25-41%) and G beta (10-33%) protein subunits. In heroin- and methadone-dependent rats naloxone (2 mg/kg)-precipitated withdrawal (2 h) did not modify the up-regulation of these G proteins induced by chronic mu-opiate treatment. In marked contrast to mu-agonists, chronic treatment with high doses of pentazocine and spiradoline or acute treatment with [D-Pen2, D-Pen5] enkephalin did not result in up-regulation of these G protein subunits. After chronic treatment with mu-agonists, significant negative correlations were found when the percentage changes in immunoreactivity of PKC-alphabeta were related to the percentage changes in immunoreactivity of G alpha(i1/2), (r = -0.53, n = 29) and G beta (r = -0.41, n = 24) in the same brains. PKC-alphabeta abundance did not correlate significantly with the density of G alpha(o) (r = -0.21, n = 28). Together the results indicate that the brain PKC-alphabeta system may play a major regulatory role in opiate tolerance and dependence. Moreover, the possible in vivo cross-communication between this regulatory enzyme and specific inhibitory G proteins may also be of relevance in the cellular and molecular processes of opiate addiction.
Ventayol
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Department of Biology, University of the Balearic Islands, Palma de Mallorca, Spain.
Busquets
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Garcia-Sevilla
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JA
D000818:Animals; D001921:Brain / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D019204:GTP-Binding Proteins / Q000187:drug effects*; D007150:Immunohistochemistry; D008297:Male; D008691:Methadone / Q000494:pharmacology; D009020:Morphine / Q000494:pharmacology*; D009294:Narcotics / Q000494:pharmacology*; D011493:Protein Kinase C / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D009294:Narcotics; D009020:Morphine; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D008691:Methadone