Interactions of cerebral serotonin and catecholamines

It has been demonstrated that the administration of large doses of dopa or 5-hydroxytryptophan (5-HTP) affected not only dopamine (DA) or serotonin (5-HT) neurons but also other neurons. This report concerns the effects of tryptophan, 5-HTP or dopa on concentrations of 5-HT, DA, norepinephrine, and their metabolites in p-chlorophenylalanine-, 5,7-dihydroxytryptamine (5,7-DHT)- or non-treated rat brain. The results were as follows: 1) When the brain DA levels increased more than six-fold after the i.p. administration of l-dopa, the brain 5-HT levels significantly decreased, and 5-hydroxyindoleacetic acid (5-HIAA) levels significantly increased. 2) In the nerve ending fraction of rat brain, the administration of l-dopa induced the largest increase of DA levels in the striatum, while administration of dl-5-HTP produced the largest increase of 5-HT levels in the hypothalamus. These results suggest that the distribution of newly synthesized 5-HT or DA after the administration of 5-HTP or dopa, respectively, depends not only on that of aromatic l-amino acid decarboxylase, but also on that of nerve cells which selectively take up 5-HTP or dopa or that of synaptic vesicles which selectively take up 5-HT or DA. 3) The large doses of 5-HTP administration to 5,7-DHT pretreated rats induced the largest increase of 5-HT levels in the striatum. Thus, results in the present study indicate that the distribution of DA or 5-HT after the administration of dopa or 5-HTP, respectively, depends on the doses of dopa or 5-HTP given as well as the presence or absence of the lesioned 5-HT or DA neurons.     

Role of catecholamines and serotonin receptor subtypes in nefopam-induced antinociception.

The non-opiate analgesic nefopam has been shown to inhibit monoamines uptake, but little is known about receptor subtypes effectively involved in its analgesic effect. In vitro binding assays yielded the following measures of affinity (IC(50)): serotonergic 5-HT(2C) (1.4 microM), 5-HT(2A) (5.1 microM), 5-HT(3) (22.3 microM), 5-HT(1B) (41.7 microM), 5-HT(1A) (64.9 microM), adrenergic alpha(1) (15.0 microM) and dopaminergic D(1) (100 microM). Subcutaneous nefopam administration dose-dependently inhibited pain in acetic acid-induced writhing (1-30 mg kg(-1)) and formalin (1-10 mg kg(-1)) tests in the mouse. Pretreatments with adrenergic alpha(1) (prazosin) and alpha(2) (yohimbine), and serotonergic 5-HT(1B) (GR127935) receptor antagonists significantly increased the nefopam ED(50) in the writhing test. The serotonergic 5-HT(2C) (RS102221) and the dopaminergic D(2) (sulpiride) receptor antagonists inhibited nefopam antinociception in the formalin test. However, in both tests, nefopam analgesic activity was not modified by the following receptor antagonists: dopaminergic D(1) (SCH23390), serotonergic 5-HT(1A) (NAN-190, WAY100635), 5-HT(2A) (R96544, ketanserin), 5-HT(3) (tropisetron), and 5-HT(4) (SDZ205557). In conclusion, nefopam analgesic activity could be modulated by the adrenergic alpha(1) and alpha(2) receptors, the dopaminergic D(2) receptors, and the serotonergic 5-HT(1B) and 5-HT(2C) receptor subtypes.     

Involvement of serotonin in zimelidine-induced hyperglycemia in mice

Effects of a selective serotonin reuptake inhibitor, zimelidine, on plasma glucose was studied in mice. Zimelidine dose-dependently induced hyperglycemia, although it did not change insulin levels. To determine the involvement of the serotonergic system in zimelidine-induced hyperglycemia, effects of the 5-HT depleter p-chlorophenylalanine(pCPA) were examined. pCPA significantly reduced zimelidine-induced hyperglycemia. This suggests that zimelidine-induced hyperglycemia is mediated by the serotonergic system through its 5-HT reuptake inhibition.     

Involvement of catecholamines in acute tolerance to ethanol in mice

The percentage of mice able to remain on a rolling drum for 45 s was recorded at 1.25 min and 30 min after administration of ethanol (2.4 g/kg). Though there was no significant difference in brain ethanol levels at the two test times, performance was markedly different with significantly fewer mice able to remain on the drum at 1.25 min than at 30 min. This phenomenon, known as acute tolerance, was antagonised by pretreating mice with haloperidol (0.4 mg/kg), FLA-63 (25 mg/kg), diethyldithiocarbamate (400 mg/kg), phenoxybenzamine (40 mg/kg), phentolamine (20 mg/kg), yohimbine (3 mg/kg) and clozapine (1 mg/kg), but not by spiperone (0.16 mg/kg), -methyl-p-tyrosine (300 mg/kg) or phenobarbitone (10 mg/kg). The relative potencies of the effective blocking agents suggest that ₂-receptors may play an important role in mediating acute ethanol tolerance.     

Involvement of catecholamines in kindled amygdaloid convulsions in the rat.

The seizure state induced by amygdaloid kindling in the rat was accompanied by a significant depletion of norepinephrine (NE) in the hippocampus, midbrain, limbic lobes and frontal cortex. In contrast, there was no change in NE levels of the hypothalamus, brain stem or basal ganglia, nor were the levels of dopamine affected in any of these brain regions. Drugs which impair central noradrenergic mechanisms, including α-methyl-p-tyrosine, disulfiram and propranolol, enhanced the rate of development of kindled seizures; disulfiram and propranolol also increased the duration of after discharges accompanying the seizures. In contrast, drugs affecting dopaminergic mechanisms, including pimozide and apomorphine, as well as drugs acting on α-adrenergic receptors, phenoxybenzamine and clonidine, had no influence on the seizure state. It appears that central noradrenergic mechanisms, particularly those involving β-adrenergic receptors, may be involved in the pathogenesis of amygdaloid kindled seizures.     

Endogenous catecholamines during noradrenaline and adrenaline hypertension.

Intravenous infusion of noradrenaline (NA) (30 micrograms/kg) results in reflectory increase of adrenaline (A), this has been established by spectrofluorometrical assay of the difference in the plasma catecholamine level during noradrenalinemia with and without elimination of circulatory reflexes. Analogically, intravenous infusion of A brings about reflectory increase of NA level. In the presence of hexamethonium a substantial amount of injected NA remains in the blood. After adrenalectomy the uptake of injected catecholamines is greatly enhanced.     

Aggressive behavior and brain serotonin and catecholamines in ants (Formica rufa).

Serotonin, adrenaline and noradrenaline were analyzed in brains of the ants, Formica rufa. Experiments demonstrated that concentrations of both serotonin and adrenaline were higher in ants that displayed aggressiveness (interspecific and intrageneric) while concentrations of noradrenaline was decreased.     

Serotonin-binding proteins in the bovine cerebral cortex: interaction with serotonin and catecholamines

The soluble serotonin-binding proteins (SBP) present in bovine frontal cortex are very similar to those reported in rat brain. Binding of [³H]serotonin to SBP, present in ammonium sulphate-precipitated proteins from bovine cortex, requires Fe²⁺ but not Fe³⁺. In the presence of an optimal concentration of Fe²⁺ (0.1 mM), bovine SBP behave as a single class of non-cooperative sites for [³H]serotonin binding (B_max = 120 ± 12 pmol/mg protein, K_D = 0.12 ± 0.04 μM, n = 3). Binding of [³H]serotonin is decreased by nucleotides and by reagents which modify sulfhydryl groups and reduce disulfide bonds and by metal ion chelators. Serotonin analogs possessing an hydroxyl group on the indole ring and catecholamine analogs possessing an intact catechol moiety are effective competitors (K_i from 0.1 to 0.3 μM). In both cases, the aliphatic amino group does not contribute to the binding, but the affinity is strongly decreased if aromatic hydroxyl groups are methoxylated. Catecholamine-SBP interactions can also be demonstrated directly by binding experiments. Binding of [³H]dopamine is greatly enhanced by Fe²⁺, Cu²⁺ and Mn²⁺, but not by Fe³⁺. The Fe²⁺-dependent binding component of [³H]dopamine is saturable (B_max = 279± 64 pmol/mg protein, K_D = 0.19 ± 0.02 μM, n = 3), and possesses the same physicochemical properties as SBP: it elutes immediately after the void volume on a Sephacryl S100 HR (1.6 × 140 cm) gel filtration column (reflecting aggregation) and it migrates with an apparent molecular weight of 57–58 kDa on native polyacrylamide gel electrophroresis. Whereas the serotonin-storing role of SBP in serotonergic neurons has already been well documented, the present data advocate that these proteins may also possess catecholamine-storing properties.     

Correlative changes of serotonin and catecholamines with pharmacokinetic alterations of imipramine in rat brain

Rats were given i.p. imipramine (20 mg/kg), acutely or chronically, and the levels of serotonin (5-HT), norepinephrine (NE), dopamine (DA) and their metabolites in the brain at different times were compared with the concentrations of imipramine and desipramine. The levels of 5-HT, DA, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the brain did not appear to be affected by quantitative alterations in the concentrations of imipramine and desipramine. The level of 5-hydroxyindole acetic acid (5-HIAA) was reduced and the level of 3-methoxy-4-hydroxyphenylglycol (MHPG) tended to decrease 3 h after imipramine administration in acutely treated rats. The reduced level of 5-HIAA was maintained during the chronic treatment with imipramine, whereas the MHPG level increased and the NE level decreased. The decrease in 5-HIAA depended on the concentration of imipramine in the brain, whereas the changes in the levels of NE and MHPG appeared to be caused by desipramine. The present studies show that pharmacokinetic variations of imipramine in the brain might correlate with the altered levels of 5-HIAA, NE and MHPG.     

Role of serotonin and catecholamines in brain in the feeding suppressant effect of fluoxetine.

The hypophagic effect of fluoxetine was studied in rats, injected intracerebroventricularly with 150 micrograms/20 microliters 5,7-dihydroxytryptamine, to destroy serotonin-containing neurones or 250 micrograms/20 microliters 6-hydroxydopamine, to destroy catecholamine-containing neurones. The effect of various serotonin receptor antagonists was assessed as well. Neither neurotoxin significantly modified the effect of 20 mg/kg (i.p.) fluoxetine on food intake. Metergoline (1-5 mg/kg), (-)-propranolol (16 mg/kg) and ICS 205-930 (0.1 and 1 mg/kg) did not modify the hypophagic effect of fluoxetine, while mianserin (1 and 5 mg/kg), ritanserin (0.5 and 1 mg/kg) and xylamidine (3 mg/kg) slightly but significantly reduced it. While the mechanism by which some 5-HT receptor antagonists modify the effect of fluoxetine remains to be elucidated, it seems clear that 5-HT receptors hardly have any significant role in the ability of the drug to suppress food intake.     

The effect of yohimbine on the turnover of brain catecholamines and serotonin.

Yohimbine moderately increased the depletion of brain dopamine (DA) after alpha-methyl-p-tyrosine (AMT) only when the two drugs were given at the same time; the baseline concentration of brain homovanillic acid (HVA) and its accumulation after probenecid were strongly increased by yohimbine. Yohimbine markedly decreased the concentration of brain noradrenaline (NA), both when given alone and before or at the same time as AMT; when it was given at an increasing interval after AMT, the effect became progressively smaller. The baseline concentration of brain 3-methyoxy-4-hydroxyphenylethyleneglycol sulphate and its accumulation after probenecid were increased by yohimbine; this effect was not as marked as that on HVA and was proportional to the quantity of NA depleted in the Amt method. The accumulation of brain k-hydroxyindoleacetic acid after probenecid was decreased by yohimbine pretreatment.     

Blood pressure and catecholamines following exercise during selective beta-blockade in hypertension

Summary This study examines and compares the hemodynamic and sympathoadrenal response to bicycle exercise in hypertensive subjects during two weeks' treatment with a cardio-selective (metoprolol) and nonselective (propranolol) beta-blocker. The increase in plasma norepinephrine and epinephrine concentration following exercise was augmented to a similar degree with each beta-blocker. Pre-exercise blood pressure and heart rate were similar for the two drugs. However immediately after exercise and particularly after resting for 20 min post exercise, diastolic blood pressure was lower during metoprolol treatment. Systolic blood pressure was also lower 20 min post exercise during metoprolol treatment. These observations indicate that cardio-selective beta-blockers offer advantages in blood pressure control during exercise through intact vascular ₂-adrenoceptors opposing sympathetically mediated vasoconstriction.     

Serotonin-binding proteins in the bovine cerebral cortex: interaction with serotonin and catecholamines.

The soluble serotonin-binding proteins (SBP) present in bovine frontal cortex are very similar to those reported in rat brain. Binding of [3H]serotonin to SBP, present in ammonium sulphate-precipitated proteins from bovine cortex, requires Fe2+ but not Fe3+. In the presence of an optimal concentration of Fe2+ (0.1 mM), bovine SBP behave as a single class of non-cooperative sites for [3H]serotonin binding (Bmax = 120 +/- 12 pmol/mg protein, KD = 0.12 +/- 0.04 microM, n = 3). Binding of [3H]serotonin is decreased by nucleotides and by reagents which modify sulfhydryl groups and reduce disulfide bonds and by metal ion chelators. Serotonin analogs possessing an hydroxyl group on the indole ring and catecholamine analogs possessing an intact catechol moiety are effective competitors (Ki from 0.1 to 0.3 microM). In both cases, the aliphatic amino group does not contribute to the binding, but the affinity is strongly decreased if aromatic hydroxyl groups are methoxylated. Catecholamine-SBP interactions can also be demonstrated directly by binding experiments. Binding of [3H]dopamine is greatly enhanced by Fe2+, Cu2+ and Mn2+, but not by Fe3+. The Fe(2+)-dependent binding component of [3H]dopamine is saturable (Bmax = 279 +/- 64 pmol/mg protein, KD = 0.19 +/- 0.02 microM, n = 3), and possesses the same physicochemical properties as SBP: it elutes immediately after the void volume on a Sephacryl S100 HR (1.6 x 140 cm) gel filtration column (reflecting aggregation) and it migrates with an apparent molecular weight of 57-58 kDa on native polyacrylamide gel electrophoresis. Whereas the serotonin-storing role of SBP in serotonergic neurons has already been well documented, the present data advocate that these proteins may also possess catecholamine-storing properties.     

Renal catecholamines and alpha 2-adrenergic receptors in salt-related and genetic hypertension

Increased dietary salt intake alters renal function which often leads to deleterious cardiovascular consequences. Studies were carried out to characterize the effects of high-salt diets on renal catecholamines and alpha 2-adrenergic receptors. These parameters were evaluated in both genetic and acquired forms of hypertension and also in normotensive rats on high-salt diets. Renal catecholamine content was determined by high-performance liquid chromatography with electrochemical detection. Renal alpha 2-adrenergic receptor-binding studies were performed on whole kidney homogenates using 3H-p-aminoclonidine to label both high- (0.5 nM) and low-affinity (5.0 nM) renal alpha 2-adrenergic receptors. Increased salt intake elevated blood pressure, decreased renal norepinephrine stores and resulted in renal alpha 2-adrenergic receptor up-regulation in deoxycorticosterone acetate salt hypertensive rats, Dahl-S rats and COX-SHR. The decreased renal stores of norepinephrine (NE) appeared to reflect increased renal NE utilization. In contrast, SHR (Charles River) had elevated NE stores and alpha 2-adrenergic receptors while on normal salt diets. Short-term (10-14 days) exposure to high-salt diets had modest effects in normotensive rats or COX-SHR, although it was sufficient to increase low affinity renal alpha 2-adrenergic receptor number. Renal dopamine metabolism was also altered by high-salt diets. These studies demonstrated a relationship between renal NE content and renal alpha 2-adrenergic receptors. The implications of this relationship and other salt-related changes in renal catecholamine metabolism were discussed as they pertained to hypertension and renal function.