Amphiphysin I but not dynamin I nor synaptojanin mRNA expression increased after repeated methamphetamine administration in the rat cerebrum and cerebellum.

Dopamine increases/decreases synaptic vesicle recycling and in schizophrenia the proteins/mRNA is decreased. We isolated cDNA clone, similar to amphiphysin 1 (vesicle protein) mRNA from the neocortex of rats injected repeatedly with methamphetamine using polymerase chain reaction (PCR) differential display. This clone is highly homologous to the 3' region of the human amphiphysin gene. PCR extension study using a primer specific for the rat amphiphysin 1 gene and a primer located within the clone revealed that it is the 3' UTR region of the rat amphiphysin 1 gene. Furthermore, in situ hybridization revealed that amphiphysin 1 mRNA is expressed in the cerebrum, medial thalamus, hippocampus and cerebellum. In the cerebellum, amphiphysin mRNA expression was confined to upper granule cell layer. Repeated methamphetamine administration increased amphiphysin I mRNA expression in both anterior part of the cerebrum, and the cerebellum. However, the repeated administration did not alter mRNA expression of the other vesicle proteins, synaptotagmin I, synapsin I, synaptojanin and dynamin I, we conclude that the repeated administration selectively increased amphiphysin 1 mRNA expression. Thus, amphiphysin 1 does not work as synaptic recycling, but it is suggested, as a part of pathogenesis of brain tissue injury (under Ca²âº and Mg²âº devoid environment) in repeated methamphetamine-injected states, the gene regulate actin-asssembly, learning, cell stress signaling and cell polarity.

Repeated administration of methamphetamine blocked cholecystokinin-octapeptide injection-induced c-fos mRNA expression without change in capsaicin-induced junD mRNA expression in rat cerebellum.

In the cerebellum, there are numerous cholecystokinin (CCK-8)-containing fibers. Since systemic CCK-8 injection-induced anxiety (psychological stress) activates the locus coeruleus cells that send mossy fiber inputs to the cerebellum, we examined whether systemic CCK-8 injections activate the rat and mouse cerebellum. First, injections of CCK-8 were found to induce c-fos mRNA expression in a vague patchy pattern that is different from single methamphetamine-induced Zebrin band-like c-fos mRNA expression, suggesting that the CCK-8 activating mossy fibers induce gene expression differently from the dopamine-containing mossy fibers in the ventral tegmental area. Second, since CCK-8 facilitates neural activity of dopamine in the midbrain, we examined whether repeated methamphetamine administration that induced behavioral sensitization had similar effects on the cerebellar CCK system. Repeated administration of methamphetamine suppressed the CCK-8-induced c-fos mRNA expression in the rat cerebellum. Third, capsaicin injections (physical stress) into a hind limb of the rat increased junD mRNA expression with no effect on c-fos mRNA expression, and repeated methamphetamine injections had no effect on the capsaicin-induced expression of junD mRNA. Fourth, either single injection of methamphetamine or CCK-8 to mice increased c-fos mRNA expression in the locus coeruleus, and so noradrenalin, but not dopamine, might interact with CCK-8-activating system. However, we considered the possibility unlikely. Thus, we conclude that repeated methamphetamine administration though dopamine selectively inhibits the c-fos mRNA expression after CCK-8 injection in the cerebellum.

Reversal of the expression pattern of Aldolase C mRNA in Purkinje cells and Ube 1x mRNA in Golgi cells by a dopamine D1 receptor agonist injections in the methamphetamine sensitized-rat cerebellum.

The cerebellum has a parasagittal modular structure, in which Zebrin (Aldolase) positive and negative bands expressed in Purkinje cell layers alternate, and is involved in amphetamine psychosis. Administration of SKF38393, a D1 receptor agonist, reversed the behavioral sensitization of methamphetamine. In the vermis, there were the binding sites of SKF38393. In methamphetamine-sensitized rats the expression of the Aldolase mRNA positive bands move laterally in the rat vermis. We provide here the evidence that the D1 agonist injections also reversed the expression pattern of both the Aldolase mRNA in Purkinje cells and Ube (ubiquitin activating enzyme) 1x mRNA in Golgi interneurons of the sensitized rats. Thus the reverse changes in gene expression pattern in the vermis may be involved in the mechanisms of the behavioral plasticity and suggests the new treatment of drug abuse.

Reversal of methamphetamine-induced behavioral sensitization by repeated administration of a dopamine D1 receptor agonist.

Repeated intermittent administration of methamphetamine (MAP) produces an enduring hypersensitivity to the motor stimulant effect of MAP, termed behavioral sensitization. Dopamine plays a critical role in the development and expression of behavioral sensitization. Here, we investigated whether a dopamine D1 receptor agonist could reverse behavioral sensitization to MAP. Administration of MAP (1.0 mg/kg, i.p.) to rats once every 3 days for a total of 5 times (days 1-13) induced the enhancement of locomotor activity after MAP challenge (0.5 mg/kg, i.p.) on day 20, verifying the development of behavioral sensitization. The MAP-sensitized rats then received a dopamine D1 agonist, R-(+)-SKF38393 (3.0 mg/kg, i.p.), once a day for 7 consecutive days (days 21-27). Behavioral analysis on days 30 and 41 revealed that the enhanced locomotor activity was reversed by repeated R-(+)-SKF38393 administration. Moreover, repeated R-(+)-SKF38393 administration reversed the increased dopamine release in the striatum after MAP challenge on day 41. Thus, repeated administration of the dopamine D1 receptor agonist induces the reversal of established behavioral sensitization to MAP and of increased dopamine release in the striatum, lasting for at least 2 weeks. Dopamine D1 receptor agonists may be useful therapeutic agents for the treatment of psychostimulant addiction.

Selective changes in the shapes of parasagittal bands of Aldoc (Zebrin) mRNA in the rat vermis of the cerebellum after repeated methamphetamine injections.

In the cerebellum the mossy and climbing projections, which excite Purkinje cells, display a parasagittal and striped organization. These projections also excite Zebrin (aldolase C: Aldoc) parasagittally. To evaluate the possibility that external stimuli can change the organization of the bands of Aldoc mRNA, we compared the effects of repeated methamphetamine administration on the Aldoc mRNA stripes in the four transverse (anterior, central, posterior and nodular) regions of the vermis with the effects on the glutamate transporter EAAT4 (SCL1A 6) mRNA stripes. In the posterior region the injections four times daily increased the fragmentation of the Aldoc mRNA stripes. The presence of a large amount of fragmentation (forty/cerebellum slice), was accompanied with large lateral dislocations of the Aldoc mRNA stripes. In the central and nodular regions, where the size of the stripe areas decreased significantly the stripes were dislocated laterally. The dislocations of the Aldoc mRNA bands did not occur after a single methamphetamine injection and thus repeated injections were necessary to change the distributions of the lateral bands. In contrast, the distributions of the SCL1A 6 mRNA stripes did not change, even though there was mild fragmentation (six/slice) of the SLC1A 6 mRNA stripes in the anterior region and decreases in the numbers (twelve/slice) in the nodular region. We concluded that excess dopamine selectively changes the location of the Aldoc mRNA compartments in the vermis while the SLC1A 6 mRNA stripes could be changed by other inputs and thus the specific transmitter system might change the specific compartment of the cerebellum.