Modifying effects of histamine on circadian rhythms and neuronal excitability.

Histaminergic activity shows a clear circadian rhythm: high levels during the active period (in rodents at night, in monkeys and humans during the day), and low levels during the sleep period. Histamine appears to be necessary for the maintenance of the circadian rhythmicity of the adrenocortical hormone release, locomotor activity and food intake, and the sleep-wakefulness cycle. In addition, a role for histaminergic neurons in the light entrainment is implicated. In phase shift studies, histamine given centrally seems to entrain the activity rhythm in the same way as light impulses and inhibition of histamine synthesis seems to block the entrainment by light. Importantly, histamine participates in the control of arousal and may be implicated in the sleep disturbances in hepatic encephalopathy. Furthermore, evidence suggests a role for histamine in overall neuronal excitability and seizure susceptibility both in animals and humans. Thus, we conclude that histamine may exert modifying effects on circadian rhythmicity and neuronal excitability.

An acute i.c.v. infusion of leptin has no effect on hypothalamic histamine and tele-methylhistamine contents in Wistar rats.

The actions of intracerebroventricularly (i.c.v.) infused leptin on food intake, body weight and hypothalamic contents of histamine and tele-methylhistamine, the main histamine metabolite in the mammalian brain, were studied in male Wistar rats. The effect of the histamine H(1) receptor blockade on leptin-induced anorexia was also examined. It was found that leptin at the dose of 10 microg i.c.v. reduced 24-h food intake by 48% as compared with the controls (P<0.01). This leptin dose reduced feeding during 2-4 consecutive days. In spite of the marked changes in food consumption and body weight gain, leptin did not alter the hypothalamic contents of histamine and tele-methylhistamine. Furthermore, the blockade of histamine H(1) receptors by mepyramine did not attenuate the effect of leptin on feeding and body weight. The findings indicate that centrally administered leptin suppresses feeding and promotes weight loss through mechanisms that do not require the direct participation of the brain histaminergic neuron system.

Effects of the histamine H(1) receptor blocker, pyrilamine, on spontaneous locomotor activity of rats with long-term portacaval anastomosis.

To find out whether the changes in the brain histaminergic system are involved in the pathophysiology of portal-systemic encephalopathy, we examined the effects of histamine H(1) receptor blockade on spontaneous locomotor activity, feeding, and circadian rhythmicity in rats with portacaval anastomosis (PCA). Pyrilamine, an H(1) receptor blocker (15 mg/kg/day), was delivered with osmotic minipumps. Spontaneous locomotor activity was recorded for 72 hours in the open-field with an electromagnetic detector. Food intake was monitored twice daily at the end of the light (7 PM) and the dark (7 AM) phases for 3 days. Histamine H(1) receptor density in the suprachiasmatic nucleus (SCN) was examined with receptor autoradiography, employing [(3)H]pyrilamine. PCA surgery led to decreased movement time and velocity and flattened amplitude of the circadian rhythms of locomotion and feeding. In sham-operated rats, pyrilamine significantly decreased the movement time and velocity, as well as the total food consumption and completely abolished the circadian rhythmicity of locomotion. In contrast, pyrilamine increased the movement time and velocity in PCA-operated rats, particularly in the dark phase, and improved the precision of the circadian rhythms of locomotion and feeding. Histamine H(1) receptor density was not altered by PCA surgery, whereas pyrilamine treatment led to the complete blockade of H(1) receptors in both sham- and PCA-operated rats. We suggest that histaminergic imbalance has contributed to the generation and maintenance of the decreased spontaneous locomotor activity and altered circadian rhythmicity following PCA surgery in the rat, probably via an H(1) receptor-mediated mechanism.

Metoprine-induced thirst and diuresis in Wistar rats.

In the present study, the renal responses to metoprine, a histamine-N-methyltransferase inhibitor, were studied in conscious rats. Metoprine (10-20 mg kg(-1)) or vehicle were administered i.p. to male Wistar rats and the effects were followed for the subsequent 24 h. It was found that as early as 3 h after the drug administration metoprine 20 mg kg(-1) had increased water consumption and urine flow approximately 6-8-fold. The treatment decreased urine osmolality and increased free water clearance, but caused no change in plasma renin activity or plasma vasopressin concentration. In addition, a metoprine-induced elevation in the systolic blood pressure was observed during the first few hours of the experiment. During the nocturnal period of the study, glomerular filtration rate and the excretion of electrolytes did not increase in metoprine-treated rats as they did in control rats. A decrease in the release of atrial natriuretic peptide was also found. The present results show that inhibition of histamine catabolism by metoprine causes massive changes in renal functions. It seems to promote water excretion by the kidneys but, on the other hand, to reduce the excretion of electrolytes. Although the exact mechanisms, especially the role of increased blood pressure and nocturnal suppression of atrial natriuretic peptide, require further clarification, the present data suggest that renin-angiotensin system and vasopressin were not involved in these renal responses to metoprine.

Effects of intracerebroventricularly infused histamine and selective H1, H2 and H3 agonists on food and water intake and urine flow in Wistar rats.

The actions of intracerebroventricularly-infused histamine and selective histamine H1, H2 and H3 receptor agonists on food and water intake and urine flow were studied in rats. It was found that 100-800 nmoles of histamine significantly suppressed feeding. The H1 agonist 2-(3- trifluoromethylphenyl)histamine (FMPH) decreased food intake, whereas the H2 agonist dimaprit was without effect. Histamine- and FMPH-induced suppressions of feeding were attenuated by blockade of H1 but not by H2 receptors. The results clearly demonstrate that activation of brain H1 receptors decreases food intake. In subsequent studies, we found that both metoprine and thioperamide, which increase histaminergic activity through different mechanisms, also reduced food intake. This finding indicates that the brain histaminergic system is associated with feeding behavior. The same is true with body water homeostasis. Histamine caused a long-lasting diuresis. Also dimaprit and metoprine increased urine flow and the blockade of H2 receptors abolished the diuretic responses to histamine and dimaprit. On the other hand, the H3 agonist (R)-alpha-methylhistamine elicited drinking and this effect could be prevented by thioperamide pretreatment. The results imply that activation of H3 receptors predominantly provokes drinking, whereas central H2 receptors mediate the diuretic effect of histamine.

The blockade of H1 receptors attenuates the suppression of feeding and diuresis induced by inhibition of histamine catabolism.

Metoprine elevates brain histamine content by blocking the conversion of histamine to methylhistamine. It suppresses food intake, increases water intake. and induces diuresis in rats. In the present experiment, to study which receptors were involved in these metoprine-induced changes, H1, H2, and H3 receptor blockers were administered to metoprine (10 mg/kg IP)-treated rats. The food and water consumption and urine excretion were measured at 10 and 24 h after the drug administration. It was found that systemic administration of the H3 receptor antagonist, thioperamide (5 mg/kg IP), supplemented the feeding suppressive effect of metoprine. In addition to this, the H1 receptor antagonist mepyramine (20 mg/kg IP) antagonized the suppression of feeding in metoprine-treated rats, whereas the H2 receptor antagonist, ranitidine (100 mg/kg IP), had no effect. Mepyramine also decreased the diuretic response to metoprine, whereas ranitidine or thioperamide were virtually without effect. The present results show that elevation of brain histamine content by inhibiting the catabolism of histamine suppresses food intake, and this effect of metoprine can be abolished by pretreatment with antihistamines. Although the blockade of H1 receptors also attenuates the diuretic response to metoprine, further studies are needed to understand the mechanisms that mediate the effects of metoprine on water balance.

Suppression of the acquisition of conditioned avoidance behavior in the rat by 3-methylcholanthrene.

Repeated treatment with 3-methylcholanthrene (MC; 25 mg/kg body weight, i.p., two times per week, 1 month) in both male and female Wistar rats resulted in decreased performance in two sessions of a two-way active avoidance procedure. In addition, young male rats that were injected repeatedly with MC prepubertally showed diminished acquisition in conditioned avoidance behavior during both sessions. It appears that MC can alter both avoidance acquisition and retention test performance in adult male and female rats, as well as in young males. This effect was not associated with alterations in sex hormone levels. The findings of this study suggest a significant influence of MC on specific mental functions.

Modification of reproductive function in the rat by 3-methylcholanthrene.

Repeated exposure of adult female Wistar rats to 3-methylcholanthrene (MC) (25 mg kg(-1) b.w., i.p., 2xwk, 1 mo) was associated with a significant increase in estrus cycle length. In addition, an increased frequency of females with constant diestrus and abnormal cycles was observed. Young females which had been exposed to MC prepubertally or whose parents had been treated with MC before and during mating also demonstrated cycle prolongation and an increased incidence of constant diestrus and abnormal cycles. These changes in female reproductive function were not associated with measurable changes in plasma sex hormone levels. In contrast, MC exposure in adult males was associated with significant reductions in circulating plasma testosterone levels. The present data also suggest that the offspring of parents who had been exposed repeatedly to MC before and during mating are also affected. Although the central nervous system in offspring of MC-treated parents appeared to be intact, their oral body temperature was significantly lower.

Fluid balance in rats of three different strains after inhibition of histamine catabolism.

The effect of metoprine, an inhibitor of histamine (HA) catabolism, on fluid balance was studied in Wistar (W) and Long-Evans (LE) rats. AVP deficient Brattleboro (BB) rats were used to evaluate which phenomena were AVP-related. W and LE rats were quite different: LE rats were "dry" rats, they drank less, had higher plasma AVP, smaller urine volume and excreted more AVP, and responded less to salt loading and water deprivation. Furthermore, LE and W rats responded differently to metoprine. When water was provided as drinking fluid, metoprine increased water intake and urine flow in W rats, but these changes were not significant in LE rats. In contrast, when the rats drank saline, urine output and saline consumption were similarly decreased in LE and W rats. Although no metoprine-induced changes in plasma AVP were observed, urinary excretion of AVP per 24 h was reduced in metoprine treated rats. Inhibition of HA catabolism by metoprine caused only minor changes in fluid balance of AVP deficient BB rats. The results show that significant differences in fluid balance can exist between rat strains and that increased availability of HA after IP given metoprine strongly affects body fluids in normal rats, especially those of the W strain. The results provide further support to the involvement of HA in the regulation of fluid balance, but to obtain a more complete picture, other factors, such as atrial natriuretic peptide, should be studied.

Metoprine, an inhibitor of histamine N-methyltransferase but not catechol-O-methyltransferase, suppresses feeding in sated and in food deprived rats.

Metoprine is a histamine N-methyltransferase (HMT) inhibitor often used to elevate endogenous histamine (HA) levels when studying the role of brain HA. Since central histaminergic systems may be involved in the regulation of feeding, the effect of metoprine on food intake was studied in sated and in food deprived rats. The treatment caused a dose-dependent decrease in food intake in sated rats. It also suppressed deprivation-induced feeding. To clarify the specificity of the treatment, the effect of metoprine on another methylating enzyme, catechol-O-methyltransferase (COMT), was examined indirectly by examining the ratio of the non-methylated dopamine metabolite, dihydroxyphenylacetic acid (DOPAC) to that of its methylated product homovanillic acid (HVA). The dopamine metabolites did not change in a manner consistent with COMT inhibition, but instead a transient decrease in DOPAC levels was observed. However, the suppression of feeding is considered to be related to the metoprine-induced inhibition of brain HA catabolism and not with the changes in dopaminergic systems. Metoprine had no effect on brain concentration of serotonin (5-HT) or its metabolite 5-hydroxyindoleacetic acid (5-HIAA). The results provide further support for the role of brain HA in the control of feeding behavior.

The effect of metoprine on glucoprivic feeding induced by 2-deoxy-D-glucose.

Metoprine is a histamine N-methyltransferase inhibitor that elevates endogenous histamine (HA) levels. Because the histaminergic mechanism may be involved in the regulation of feeding behavior as well as in body glucose homeostasis, the effect of metoprine on glucoprivic feeding was studied in Wistar rats. Although metoprine treatment (10 and 20 mg/kg, IP) decreased feeding, the rats still responded to the administration of 400 mg/kg of 2-deoxy-D-glucose (2-DG) by increasing their feed intake. No difference was seen in the 6-h cumulative feed intake after administration of 2-DG between the metoprine- and solvent-treated rats. However, the response was delayed, and with 20 mg/kg metoprine the feed intake was significantly reduced during 2 h after 2-DG application. Both 2-DG and metoprine elevated plasma glucose concentration despite their opposite effects on feeding. Hypothalamic HA or its metabolite levels were not affected by 2-DG. The results suggest that the effects of metoprine and 2-DG are largely independent of each other, and that the feeding modulating function of HA is on such a level that it does not prevent the glucoprivic emergency response.

Assessment of acute behavioral toxicity of low doses of diisopropylfluorophosphate (DFP) in rats.

The effect on behavior of single subtoxic doses (100 and 600 micrograms/kg i.p., i.e. 1/77 and 1/13 of LD50, respectively) of an organophosphorous compound, diisopropylfluorophosphate (DFP), was studied in male Wistar rats. In the open-field test, the lower dose of DFP tended to increase ambulation, while the higher dose showed a trend towards a decrease in ambulation, rearing and frequency of defecation. In the elevated plus-maze, rotarod, elevated bridges and hot plate tests, DFP-treated rats did not differ significantly from the olive oil-treated controls. DFP significantly impaired the performance of rats in the one-trial passive avoidance task and dose-dependently decreased spontaneous locomotor activity for 4 hours after administration. At the doses used DFP only slightly inhibited acetylcholinesterase activity in the blood and different brain areas. The results show that the higher dose of DFP had an inactivating effect on the behavior of rats, while the lower dose did not markedly change their behavioral pattern. Our findings indicate that anticholinesterase compounds, such as DFP, can alter behavior even after single small subtoxic doses.

Acute behavioural effects of the organophosphates sarin and soman in rats.

The effects on behaviour of single subtoxic doses of two potent organophosphorous compounds, sarin (isopropyl methylphosphonofluoridate, 12.5 and 50 micrograms/kg, intraperitoneally) and soman (pinacolyl methylphosphonofluoridate, 4 and 20 micrograms/kg, intraperitoneally) were studied in male Wistar rats. In the open field test, soman dose-dependently decreased rearing and ambulation and increased non-mobile exploration. The higher dose of sarin changed only the rearing and grooming behaviour. Sarin and soman decreased locomotor activity on the Animex for at least one hour at the beginning of the monitoring period. In the doses used, both organophosphates inhibited acetylcholinesterase (AChE) activity significantly in the blood. The results suggest that small doses of sarin and soman have inactivating effects on the behaviour of rats. Although the findings cannot be extrapolated directly to behavioural changes in man, they indicate that subtle behavioural dysfunctions could also occur in humans at exposures which do not cause acute toxicity.

Feed intake after inhibition of histamine catabolism.

It has been reported that histamine and its precursor histidine have a feeding-suppressing effect. The present study shows that metoprine (20 mg/kg i.p.), which increases brain histamine levels by inhibiting its catabolism, also significantly decreases daily feed intake in rats. The reduced feed consumption is evident in different states of water balance. These results agree with an involvement of histaminergic systems in the regulation of feeding behaviour.

Tremorigenic effect and inhibition of tryptamine and serotonin receptor binding by beta-carbolines.

The abilities of some naturally occurring beta-carbolines (BCs), dihydro-BCs and tetrahydro-BCs to inhibit the specific binding of 3H-tryptamine (TA), 3H-serotonin (5-HT) and 3H-ketanserine to rat brain membranes and to induce tremor in mice were studied. These compounds, particularly DHBCs and BCs, showed higher affinity for TA binding sites than to 5-HT1 or 5-HT2 binding sites inhibiting the former at nanomolar and the two latter ones at micromolar or high micromolar concentrations. The Ki values for norharmane, harmaline and harmine (17, 18 and 74 nM, respectively) for TA sites indicate the highest affinity so far described for natural beta-carbolines to any receptor sites and thus may indicate their major site of action. among the BC derivatives studied, the before mentioned harmala alkaloids were the most potent inducers of tremor in mice, although the orders of the tremorogenic potency and the binding to TA site did not correlate. It is suggested that especially the tremorigenic effect of BC derivatives is partly based on the binding to specific tryptamine receptors.