Differential distribution of synapsin IIa and IIb mRNAs in various brain structures and the effect of chronic morphine administration on the regional expression of these isoforms.

Quantitative reverse transcriptase-polymerase chain reaction and in situ hybridization techniques were used to determine the regional distribution of synapsin IIa and IIb mRNAs in rat central nervous system and to assess the effect of chronic morphine administration on the gene expression of these two isoforms of synapsin II. These isoforms are members of a family of neuron-specific phosphoproteins thought to be involved in the regulation of neurotransmitter release. Our data demonstrate the widespread distribution, yet regionally variable expression, of synapsin IIa and IIb mRNAs throughout the adult rat brain and spinal cord. The ratios of the relative abundance of synapsins IIa and IIb differed by up to 4.5-fold among the various regions studied. Synapsin IIa and IIb mRNAs were shown to be highly concentrated in the thalamus and in the hippocampus, whereas lower concentrations were found in most other central nervous system structures. In this study, we show differential regulation by morphine of synapsins IIa and IIb in various regions of the brain. In the striatum, a 2.4-fold increase was observed in the levels of synapsin IIa mRNA following chronic morphine regime, whereas no change was found for synapsin IIb. On the other hand, mRNA levels of synapsin IIb in spinal cord of chronically treated rats were markedly decreased (by 62%), while no alterations were observed in synapsin IIa. Selective regulation by morphine has also been demonstrated in several other central nervous system structures. The opiate-induced regulation of the gene expression of synapsin II isoforms could be viewed as one of the cellular adaptations to the persistent opiate effects and may be involved in the molecular mechanism underlying opiate tolerance and/or dependence.

Stress- and yohimbine-induced release of cholecystokinin in the frontal cortex of the freely moving rat: prevention by diazepam but not ondansetron.

The in vivo release of cholecystokinin (CCK)-like material (CCKLM) was measured in the frontal cortex of freely moving rats using the microdialysis technique combined with a sensitive radioimmunoassay. Local perfusion of K+ (100 mM)-enriched artificial CSF resulted in a 10-fold increase in CCKLM outflow, as compared with that occurring under basal resting (K+ = 3.0 mM) conditions, and this effect could be completely prevented by removal of Ca2+ in the perfusing fluid. Chromatographic analyses demonstrated that CCK-8S contributed to 70% of CCKLM. Stressful stimuli such as a 2-min exposure to diethyl ether and a 30-min restraint produced a marked but transient increase in cortical CCKLM release. In addition, anxiety-like behavior induced by the systemic administration of yohimbine (5 mg/kg i.p.) was associated with a long-lasting enhancement in the peptide outflow. Pretreatment with the potent anxiolytic drug diazepam (5 mg/kg i.p., 5 min before each condition), which exerted no effect on its own, completely prevented CCKLM overflow due to diethyl ether, restraint, or yohimbine administration. In contrast, neither the systemic injection (0.1 mg/kg i.p.) nor the local application (100 microM through the microdialysis probe) of the serotonin 5-HT3 antagonist ondansetron affected the increased release of CCKLM in rats restrained for 30 min or treated with yohimbine. These results indicate that cortical CCKergic neurotransmission is increased during stress or anxiety-like behavior in rats. Prevention of this effect by diazepam suggests that an inhibitory influence of benzodiazepines on cortical CCKergic neurons might participate in the anxiolytic action of these drugs.