Melatonin: effects on dopaminergic and serotonergic neurons of the caudate nucleus of the striatum of male Syrian hamsters.

The effects of daily late afternoon administration of the indoleamine, melatonin, on the in situ activity of tyrosine hydroxylase (TH) and tryptophan hydroxylase (TPH) were examined in the caudate nuclei of the striatum of male Syrian hamsters. TH and TPH activities were determined in tissue extracts by measuring the accumulation of L-Dopa and 5-HTP respectively, following the administration of the aromatic L-amino acid decarboxylase inhibitor, NSD-1015. Animals were sacrificed at 4 time points over the 24 light/dark cycle after 9.5 weeks of melatonin treatment. TH activity was significantly increased by melatonin during the early part of the dark phase of the light/dark cycle. While no significant effects of melatonin on TPH was observed, melatonin significantly increased 5-HT concentrations, suggesting a melatonin-induced inhibition of 5-HT release. The data suggest that the striatum may be a region in which dopaminergic neurons are subject to significant regulation by melatonin, either directly or through serotonergic neurons which synapse on dopaminergic neurons in the striatum.

Lamotrigine inhibits the in situ activity of tyrosine hydroxylase in striatum of audiogenic seizure-prone and audiogenic seizure-resistant Balb/c mice.

Lamotrigine (LTG), an anticonvulsive drug, was tested for its effects on striatal content of DA and its metabolites, DOPAC and HVA, in audiogenic seizure-resistant (ER) and audiogenic seizure-prone (EP) lines of Balb/c mice. A single dose of LTG (20 mg/kg) prevented audiogenic seizures in seizure-prone mice, while reducing substantially the striatal content of the DA metabolite, DOPAC (to less than 50% of saline-injected controls) in both seizure-resistant and seizure-prone mice. LTG administration also resulted in significant reduction of striatal content of HVA. The in situ activity of tyrosine hydroxylase (TH) in extracts of striatum was significantly reduced by LTG administration in both ER and EP mice. These data show that DA synthesis in the striatum of mice is substantially reduced by LTG administration.

Effects of valproate on amino acid and monoamine concentrations in striatum of audiogenic seizure-prone Balb/c mice.

The effects of valproate on CNS concentrations of gamma-aminobutyric acid (GABA), glulamate (GLU), glutamine (GLN); dopamine (DA), serotonin (5-HT), and metabolites were examined in tissue extracts of caudate nucleus of genetic substrains of Balb/c mice susceptible (EP) or resistant (ER) to audiogenic seizures. Generalized tonic-clonic seizures observed in EP mice were inhibited by valproate, administered 1 h prior to testing, in a dose-response fashion. Concentrations of GABA, GLU, and GLN, which were lower in EP mice than in ER mice, were significantly increased by valproate at doses of 180 and 360 mg/kg. Concentrations of homovanillic acid (HVA) and hydroxyindoleacetic acid (5-HIAA), metabolites of DA and 5-HT, were substantially increased by valproate at these doses. The in situ activity of tyrosine hydroxylase (TH) was not significantly influenced by valproate, whereas a valproate-induced increase in tryptophan hydroxylase (TPH) activity was observed in both striatum and in midbrain tegmentum. The data are consistent with the interpretation that anti-convulsive doses of valproate influences the intraneuronal metabolism of monoamines, GABA, and glutamate concurrently. Valproate's influence on the metabolism of both major inhibitory (GABA) and excitatory (GLY amino acids in striatum could contribute to its anti-convulsive effects in genetically seizure prone mice, as well as to the accumulation of DA and 5-HT metabolites.

Melatonin increases the in situ activity of tyrosine hydroxylase in the mediobasal hypothalamus of male Syrian hamsters.

The effects of daily late afternoon injections of melatonin on the in situ activity of tyrosine hydroxylase (TH) were examined in the median eminence/arcuate region of the mediobasal hypothalamus (MBH) and the neurointermediate lobe (NIL) of the male Syrian hamster. TH activity was determined in tissue extracts by measuring the accumulation of L-DOPA following administration of the dopa decarboxylase inhibitor, NSD-1015. After 9 weeks of melatonin treatment, highly significant increases in the activity of MBH TH were demonstrated over a 24 hr period, compared to saline-treated controls. Melatonin-induced elevations in TH occurred concomitantly with decreases in tuberoinfundibular dopamine (TIDA) and tuberohypophyseal dopamine (THDA) concentrations. Similar findings were observed in castrated hamsters, indicating that the melatonin-induced increase in TH was not secondary to melatonin-induced changes in circulating levels of gonadal hormones. These data led to the interpretation that melatonin treatment elevated TIDA synthesis either through a direct action on the arcuate nuclei or on neurons impinging on these nuclei.

Determination of amino acids and monoamine neurotransmitters in caudate nucleus of seizure-resistant and seizure-prone BALB/c mice.

Amino acid and monoamine concentrations were examined in tissue extracts of caudate nucleus of genetic substrains of BALB/c mice susceptible or resistant to audiogenic seizures. Amino acids [aspartate, glutamate, glycine, taurine, serine, gamma-aminobutyric acid (GABA)], monoamines, and related metabolites were separated by isocratic reverse-phase chromatography and detected by a coulometric electrode array system. In situ activity of tyrosine hydroxylase and tryptophan hydroxylase were determined by measuring the accumulation of L-DOPA and 5-hydroxytryptophan after administration of the decarboxylase inhibitor NSD-1015. Highly significant decreases in concentrations of both excitatory (glutamate and aspartate) and inhibitory amino acids (GABA and taurine) were observed in extracts of caudate nucleus of seizure-prone mice. Substantial decreases in concentrations of dopamine (DA) and its metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, were also noted. Decreased accumulation of L-DOPA after NSD-1015 administration provided evidence for decreased tyrosine hydroxylase activity and decreased DA synthesis in striatum of seizure-prone mice compared with seizure-resistant mice. Decreased concentrations of the DA metabolite 3-methoxytyramine (after NSD-1015 administration) suggested that DA release was also compromised in seizure-prone mice. No significant difference in 5-hydroxytryptophan accumulation in striatum of seizure-prone and seizure-resistant mice suggested that tryptophan hydroxylase activity and serotonin synthesis were not affected. The data suggest that seizure-prone BALB/c mice have a deficiency in intracellular content of both excitatory and inhibitory amino acids. The data also raise the issue of whether GABAergic interactions with the nigrostriatal DA system are important in the regulation of audiogenic seizure susceptibility.

Melatonin reduces dopamine content in the neurointermediate lobe of male Syrian hamsters.

The effect of daily late afternoon administration of melatonin on the in situ activity of tyrosine hydroxylase (TH) was studied in the posterior pituitary (neurointermediate lobe) of the male Syrian hamster. After 3 weeks of melatonin administration, TH activity was significantly reduced in the posterior pituitary. This was associated with a significant decrease in norepinephrine (NE) content. After 5 weeks, TH activity and NE content were no longer significantly different from controls. Dopamine (DA) content of the posterior pituitary was decreased progressively by melatonin administration, with a reduction of greater than 50% after 5 weeks of treatment. These data provide evidence that melatonin has a potent inhibitory effect on the regulation of the dopaminergic system of the neurointermediate lobe--an effect that appears unrelated to changes in axonal TH.

Extrahypothalamic effects of melatonin administration on serotonin and norepinephrine synthesis in female Syrian hamsters.

The effects of daily late afternoon injections of melatonin for 10 weeks on the metabolism of serotonin (5-HT) and norepinephrine (NE) were examined in regional brain extracts of intact and ovariectomized (GX) Syrian hamsters. Accumulation of 5-HT and NE after administration of the monoamine oxidase inhibitor pargyline was used as a measure of the rate of neurotransmitter synthesis-with concentrations determined by HPLC with electrochemical detection. Daytime 5-HT synthesis was significantly decreased in the amygdala of melatonin-treated hamsters that had been GX (to 50% of GX controls). No significant effect on 5-HT synthesis could be detected in the mediobasal hypothalamus (MBH), however, a significant increase was demonstrated in the pontine brain stem of both intact and GX hamsters treated with melatonin. Daytime NE synthesis was decreased to levels not significantly different from zero in the amygdala of GX hamsters treated with melatonin, while in the brain stem, melatonin reduced NE synthesis in both intact and GX animals. The present data demonstrate that these melatonin effects on 5-HT and NE metabolism are not limited to the MBH and are not secondary to melatonin-induced changes in circulating levels of the ovarian steroids.

Effects of daily afternoon melatonin administration on monoamine accumulation in median eminence and striatum of ovariectomized hamsters receiving pargyline.

The effects of daily afternoon melatonin injections on the accumulation of monoamines were studied in extracts of median eminence, and of caudate nucleus, of intact and ovariectomized Syrian hamsters which were administered pargyline 2 h prior to sacrifice. Although no significant effect of melatonin administration on the serotonin (5HT) accumulation after pargyline could be detected, significantly increased amounts of 5HT and of the 5HT metabolite, 5-hydroxyindole acetic acid, were detected in median eminence and in caudate nucleus of melatonin-injected hamsters not treated with pargyline. In both median eminence and in posterior pituitary, dopamine (DA) concentrations were significantly reduced by melatonin administration. In the median eminence of intact hamsters, the accumulation of DA after pargyline was reduced to 22% of controls by melatonin injections; in ovariectomized hamsters, the accumulation of DA was reduced to 9% of controls by melatonin injections. The accumulation of norepinephrine after pargyline was significantly reduced by melatonin administration only in ovariectomized hamsters. No significant inhibitory effects of melatonin injections could be detected on DA accumulation in caudate nucleus. These data suggest that melatonin injections result in substantial inhibition of daytime DA synthesis in median eminence independently of its effects on gonadal steroids. Paradoxically, melatonin-induced inhibition of median eminence DA activity occurred concomitantly with suppression of pituitary and plasma prolactin (PRL). We conclude that daily afternoon melatonin injections inhibit PRL secretion and interfere with cycles of LH in spite of decreased DA activity in the median eminence.