Inhibition of protein kinase C but not protein kinase A attenuates morphine withdrawal excitation of rat hypothalamus-pituitary-adrenal axis.

Our previous studies have shown an enhanced activity of the hypothalamus-pituitary-adrenocortical axis response in rats withdrawn from morphine, which results from an increase in the hypothalamic paraventricular nucleus noradrenergic activity that is dependent on alpha-adrenoceptor activation. The first objective of this work was to examine the effect of protein kinase A (PKA) and protein kinase C (PKC) inhibitors on morphine withdrawal-induced changes in corticosterone release (an index of the hypothalamus-pituitary-adrenocortical axis activity) and in catecholaminergic turnover in the paraventricular nucleus. Plasma corticosterone levels as well as the concentration of noradrenaline, 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG), dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) in the paraventricular nucleus were determined. The second purpose of the study was to assess whether kinase inhibitors, administered continuously through s.c. osmotic minipumps, get into the brain. Chronic pretreatment for 7 days with the selective PKA inhibitor N-(2'guanidinoethyl)-5-isoquinolinesulfonamide (HA-1004) concomitantly with morphine did not affect the increase in corticosterone release observed after naloxone-precipitated morphine withdrawal. However, pretreatment with the selective PKC inhibitor, calphostin-C significantly antagonized the corticosterone hypersecretion in morphine-withdrawn rats. Neither HA-1004 nor calphostin-C co-administered with morphine for 7 days did modify the morphine withdrawal-induced increase in noradrenaline turnover. Pretreatment with HA-1004 inhibits the increase in dopamine turnover during morphine withdrawal, whereas calphostin-C did not affect the DOPAC/dopamine ratio. Our results might indicate that expression of morphine dependence for hypothalamus-pituitary-adrenocortical axis hyperactivity involves PKC but not PKA signaling mechanisms. It is suggested that in rats PKC may be up-regulated during morphine dependence. High-performance liquid chromatography (HPLC) analysis of hypothalamic tissue from rats perfused with kinase inhibitors demonstrates that both calphostin-C and HA-1004 can cross the blood-brain barrier when administered peripherally.