STUDIES OF RECEPTOR-MEDIATED INHIBITION OF CA-45 ACCUMULATION INTO SYNAPTOSOMES

1. The effects of α2-adrenoceptor and κ-opiate receptor activation on 45Ca accumulation into rat cortical synaptosomes were examined. 2. Clonidine (1 μM) and U50488H (1 μM) significantly reduced 45Ca accumulation under both resting (5 mM K+) and depolarizing (15-30 mM K+) conditions. 3. The inhibitory effects of the agonists on 45Ca accumulation into synaptosomes were enhanced in the presence of the Na+-Ca2+ exchange inhibitor sodium orthovanadate (vanadate, 2 mM), and were not present in mitochondrial preparations. 4. When the agonists were used together, their inhibitory effects were not additive but were, in fact, attenuated. 5. In the presence of the α2-adrenoceptor antagonist idazoxan (1 μM), the inhibitory effect of U50488H on 45Ca accumulation was enhanced. A similar increase in the inhibitory effectiveness of clonidine was observed in the presence of naloxone (20 μM). 6. When synaptosomes were pretreated with 3-isobutyl-1-methylxanthine (IBMX, 0.5 mM), dibutyryl-cyclic AMP (db-cyclic AMP, 10 μM), or 8-bromo-cyclic AMP (8Br-cylic AMP, 10 μM), the inhibitory effects of clonidine and U50488H were abolished, suggesting that a decrease in cyclic AMP production is part of the receptor-effector coupling mechanism of both receptor systems. 7. The phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA, 0.05 μM) increased 45Ca accumulation but did not alter the inhibitory effects of clonidine or U50488H, thus showing that the effects of the agonists are not mediated by protein kinase C. 8. We conclude that α2-adrenoceptor and κ-opiate receptor activation dramatically reduce 45Ca influx through Ca2+ channels (e.g., by 50%), that there is a functional antagonism between the two receptor systems and that in both cases, the receptor effector mechanism involves a decrease in cyclic AMP production.