Increases in serotonergic neuronal activity following intracerebroventricular administration of AF64A in rats.

Changes in the serotonergic nervous system after the intracerebroventricular (i.c.v.) administration of ethylcholine aziridinium (AF64A, 3 nmol/each ventricle) were studied in rats. Two weeks after the infusion of AF64A, the levels of 5-HT and 5-HIAA in microdialysed cerebrospinal fluid (CSF), the levels of total 5-HT and 5-HIAA, the density of serotonin uptake sites and the activities of tryptophan hydroxylase (TPH) and monoamine oxidase (MAO) in various brain regions were determined. After AF64A administration, the concentrations of 5-HT in lateral ventricle were increased and the levels of 5-HIAA were decreased. However, the hippocampal levels of total 5-HT were decreased without changes in the levels of 5-HIAA and the hippocampal turnover rates of 5-HT increased. Also, the density of uptake sites of serotonin ([(3)H]citalopram binding sites) was decreased in the various brain. The activities of TPH were increased in striatum and frontal cortex and the activity of MAO was also increased in striatum. These results indicate that AF64A induces an increase in serotonergic neuronal activity and decreased densities of 5-HT uptake sites which may affect the change in the other parameters of serotonergic neuronal activities. Furthermore, these results suggest that the impaired cholinergic neuronal activity induces the alteration in the serotonergic nervous activities.

Effects of dehydroevodiamine exposure on glutamate release and uptake in the cultured cerebellar cells.

Dehydroevodiamine has been reported to have neuroprotective and antiamnesic effects. This study examined the effects of dehydroevodiamine on glutamate release and uptake in cultured cerebellar cells. Chronic dehydroevodiamine exposure decreased the viability of granule cells. The basal and N-methyl-D-aspartate (NMDA)-induced release of glutamate from granule cells were decreased (26 and 14%) by dehydroevodiamine. The NMDA-induced release of glutamate was concentration-dependently inhibited in the granule cells. The basal and NMDA-induced releases of glutamate in chronically dehydroevodiamine-preexposed granule cells were unaffected by dehydroevodiamine. Glutamate uptake in the glial cells incubated without and with cAMP was inhibited (31% and 8%, respectively) by dehydroevodiamine. In the chronically dehydroevodiamine-preexposed glial cells, glutamate uptake was increased (8%) in the cAMP-coexposed glial cells by dehydroevodiamine but was unaffected in the naive cells. In addition, dehydroevodiamine potentiated (from 20% to 34%) the inhibition of L-pyrollidine-2,4-dicarboxylic acid (PDC) on glutamate uptake in naive glial cells, but this inhibition was reduced (from 41% to 26%) in cAMP-coexposed glial cells. These results suggest that dehydroevodiamine inhibits glutamate uptake and release. Furthermore, the results suggest that the characteristics of glutamate release and uptake in granule and glial cells may be altered by chronic exposure to dehydroevodiamine.

Opposite modulation of glutamate uptake by nicotine in cultured astrocytes with/without cAMP treatment.

Cultured cerebellar astrocytes were exposed to nicotine with or without dibutyryl cAMP (dBcAMP) for 2 to 10 days in situ in order to determine the effects of nicotine exposure on glutamate uptake in differently conditioned astrocytes. After acute nicotine exposure, glutamate uptake was lower and higher in the naive and the preconditioned astrocytes, respectively. After subacute nicotine exposure, the inhibitory effect of L-trans-pyrollidine-2,4-dicarboxylic acid (PDC) on the glutamate uptake in the naive and the conditioned cells was decreased and increased, and the IC50's for PDC were higher and lower, respectively. In addition, glutamine synthetase activity after subacute nicotine exposure was lower in the naive and higher in the conditioned astrocytes; the change in Na+/K+ ATPase activity was the opposite to that in glutamine synthetase activity. These results suggest that nicotine modulates the characteristics of glial glutamate transporters and glutamine synthetase activity in astrocytes. Furthermore, nicotine might differently affect the state of glial cells during development.

Chronic exposure of nicotine modulates the expressions of the cerebellar glial glutamate transporters in rats.

Rats were given nicotine (25 ppm) in their drinking water at the start of their mating period in order to study the expressions of glutamate transporter subtypes in cerebellar astrocytes following the chronic exposure of nicotine after mating. After the offspring were delivered, each group was divided into two subgroups. One group, the control group, was given distilled water only and the other group, the experimental group, was given distilled water containing nicotine. The cerebellar astrocytes were prepared from 7 day-old pups at each group. Ten days after the cells were cultured, the expression of the glutamate transporter subtypes (GLAST and GLT-1) was determined using immunochemistry and immunoblotting. During the continuous treatments, the developmental expression patterns of the GLAST and GLT-1 in the cerebellum were also determined from 2, 4 and 8 week-old rats. The expression levels of GLAST in cultured astrocytes of both the pre- or post-natally exposed groups were higher than those of the control group. However, these expression levels of the continuously exposed group were lower than those of the control group. Compared to those of the control group, the GLT-1 expression levels of all the nicotine-treated groups were higher, particularly in the continuously treated group. According to the results from the immochemistry procedure, the cerebellar GLAST and GLT-1 expression levels of all nicotine-treated groups were lower than those of the control group at each age. However, the immunoblotting procedure showed that the cerebellar GLT-1 expression levels of all the nicotine-treated groups were higher than those of the control group, except for the rats that were continuously exposed for 8 weeks using immunoblotting. These results suggest that the expression of the glial GLAST and GLT-1 are altered differently depending on the initial exposure time and the particular period of nicotine exposure. In addition, nicotine exposure during gestation has persistent effects on glial cells.