Action of cisapride on rat colonic secretion

Summary The stimulating effect of cisapride on the motility of the digestive tract is well known. However, there are only a few studies on the influence of this drug on the absorptive or secretory activity of the colonic mucosa. In the present study, the ability of cisapride to alter the mural transport of water and electrolytes in the colon and its effects on mucus secretion and albumin permeation were studied.The effects of cisapride on the rat colon in vivo were studied under different conditions, by means of an instillation of sodium acetate solution at pH 6.9, which induced absorption of water and electrolytes, and in two models of colonic secretion, one employing the instillation of an acetic acid solution at pH 2.9 and the other, an intravenous infusion of 5-hydroxytryptamine (serotonin) 45 g · kg^–1 · min^–1 together with intracolonic instillation of sodium acetate.Cisapride (i.v.), at a dose of 0.32 mg · kg^–1, in rats whose colon was instilled with sodium acetate (pH 6.9) induced an increase in sodium absorption and a reduction in water absorption.Cisapride (iv.), at doses of 0.32, 0.64 and 1.0 mg · kg^–1, inhibited the secretion of water, Na⁺, Cl^–, and mucus and the permeation of albumin induced by acetic acid instillation or by serotonin infusion. It is concluded that the effect of cisapride on the colonic mucosa varies in accordance with the functional mucosal conditions and that this action may be of clinical importance. Correspondence to D. Celener at the above address     

The effect of phloxin on bethanechol and histamine stimulated gastric acid secretion in rats

Summary The effect of phloxin (Na salt of tetrabromo-tetrachlorofluorescein) on histamine- and bethanechol-stimulated gastric acid secretion was studied in anaesthetized rats. Concentrations of phloxin, equimolar with those of the stimulants, depressed the secretory response to histamine and had no influence on the bethanechol effect. The results suggest that histamine is not involved in the events triggered by bethanechol which stimulate gastric acid secretion in rats.Supported by the Deutsche Forschungsgemeinschaft and the Gesellschaft der Freunde der Universität Tübingen.     

Reversal of the bronchodilator action of ephedrine during ventilation with carbon dioxide

Ephedrine reduces the bronchoconstrictor effects of histamine, 5-hydroxytryptamine and acetylcholine in guinea-pigs ventilated with air, but during ventilation with 10% carbon dioxide in air, when blood pH is lowered and blood P_CO2 is raised, ephedrine enhances the bronchoconstrictor responses to all three drugs. Pretreatment with monoamine oxidase inhibitors did not affect the bronchodilator actions of ephedrine, but significantly increased its action in enhancing bronchoconstrictor responses during ventilation with 10% carbon dioxide in air. It is suggested that ephedrine sensitizes bronchial smooth muscle to constrictor drugs and this effect is manifested when the bronchodilator action of ephedrine is abolished. The findings suggest a potentially hazardous effect of ephedrine in the treatment of severe asthmatic attacks.     

Interaction between 5-hydroxytryptamine and other vasoconstrictor substances in the isolated femoral artery of the rabbit; effect of ketanserin (R 41 468)

Experiments were designed to determine whether or not ketanserin (R 41 468) antagonizes the augmentation by 5-hydroxytryptamine of contractions evoked in isolated arteries by non-adregergic vasoconstrictor substances. Rings of rabbit femoral arteries were studied under isometric conditions in organ chambers filled with Krebs-Henseleit solution (37°C). Ketanserin, unlike methysergide and LSD, was devoid of agonistic properties. It competitively antagonized contractile responses to 5-hydroxytryptamine and, at higher concentrations, to histamine. 5-Hydroxytryptamine amplified the contractions evoked by threshold concentrations of histamine, angiotensin II and prostaglandin F_2α; in all three cases, the amplification was antagonized by comparable concentrations of ketanserin. These experiments indicate that the interaction of 5-hydroxytrptamine with S₂-receptors of the vascular smooth muscle cells is essential to allow the expression of the monoamine-induced amplification of the response to other vasoconstrictor substances. The inhibition by ketanserin of the amplifying effect of 5-hydroxytryptamine on vascular responses may help explain the antihypertensive properties of the compound.     

Inhibition by 5-hydroxytryptamine of anaphylactic histamine release from bovine granulocytes.

A suspension of granulocytes from calves sensitised to horse plasma released histamine when exposed to the antigen. Histamine release was strongly inhibited by 5-hydroxytryptamine in a dose-related fashion. 5-HT is known to be released from bovine lungs in vitro, and may regulate the release of histamine in vivo during anaphylaxis.     

Activity of 2-(5-methyl-4-imidazolyl)-ethylamine (5-methylhistamine) on the gastric secretion of different laboratory animals and of man

The compound 2-(5-methyl-4-imidazolyl)-ethylamine, namely 5-methylhistamine3(5-MeH), was compared to histamine for its stimulant activity on gastric secretion in Heidenhain pouch dogs, cats, rats and also on gastric secretion of human volunteers.In dogs and cats 5-MeH had approximately the same activity as histamine, both as regards threshold doses and peak secretory effects, when given by subcutaneous injection or by intravenous infusion.In the rat 5-MeH was 60% less active than histamine when given by either i.v. injection or infusion.In the preliminary experiments carried out in man, 5-MeH and histamine given at the same doses had approximately the same effect as regards volume, acidity of the gastric juice and also persistence of effect. 5-MeH was tolerated in all the subjects whereas histamine was not tolerated in 1 out of 5 subjects.The results obtained suggest that this new methyl derivative could reprent a good alternative to thmmon stimulants used in the clinical gastric secretory tests.     

The effects of intra-arterial bradykinin, histamine, acetylcholine and prostaglandin E1 on nociceptive and non-nociceptive dorsal horn neurones of the cat.

The effects of peripherally administered algesic agents were investigated on the firing of cat dorsal horn interneurones classified as nociceptive or non-nociceptive according to the peripheral stimuli that excited them. A small amount of bradykinin injected into the blood supplying the receptive fields of cells was a potent specific stimulus causing activation of nociceptive cells and slowly conducting nerve fibres. Larger amounts of bradykinin and large amounts of histamine, 5-hydroxytryptamine and acetylcholine activated both nociceptive and non-nociceptive cells. Prostaglandin E1 enhanced the effects of bradykinin and histamine on nociceptive cells. Prostaglandin E1 also increased the response of these cells to the application of noxious heat whilst aspirin reduced this response. These results support a chemosensitive theory of nociceptor activation and show bradykinin to be the most potent and specific of the suggested endogenous algesic agents in causing activation of CNS nociceptive pathways.     

Age-dependent suppression of hippocampal epileptic afterdischarges by metabotropic glutamate receptor 5 antagonist MTEP

Action of an antagonist of metabotropic glutamate receptors subtype 5 MTEP was studied in a model of complex partial seizures. Dorsal hippocampus of rat pups 12, 18 and 25 days old was stimulated six times with 10-min intervals. MTEP (20 or 40 mg/kg) was injected after the first afterdischarge and duration of afterdischarges was measured.MTEP exhibited marked anticonvulsant action in 12-day-old-rats, the similar effect in 18-day-old rats was observed only with the second stimulation. No anticonvulsant action was seen in 25-day-old animals.Our results may qualify antagonists of mGluR5 as potential antiepileptic drugs for some types of childhood epilepsies.     

The effect of drugs influencing amine synthesis on the analgesic action of tremorine

The effect of reserpine, -methyl-dl-m-tyrosine (-MT), diethyl-dithiocarbamate (DDC) and p-chlorophenylalanine (p-Clphe) pretreatment on tremorine induced analgesia has been studied in mice by various methods. The ED 50 of tremorine was significantly increased by all the methods when the animals were pretreated with reserpine, DDC and p-Clphe. -MT had no significant effect on tremorine induced analgesia. The results are discussed with respect to the role of brain amines in the analgesic action of tremorine.     

Development of tolerance to the stimulatory effect of morphine in dog intestine

Intestinal motor responses to morphine and 5-hydroxytryptamine (5-HT) were measured in dogs injected daily for 16 days with morphine (stabilized at 30 mg/kg i.v.) and compared to responses obtained in control animals. Intestinal contractions produced by 10 and 20 μg/kg morphine were reduced in the morphine-treated animals. Responses to 5-HT did not differ between the two groups. The contractor dose-response curve for morphine, but not for 5-HT or bethanechol, was shifted markedly to the right in intestinal segments isolated from morphine-treated dogs. It is concluded that tolerance to the intestinal stimulatory effect of morphine occurs in the dog.     

Further observations on the effect of repeated electroconvulsive shock on the behavioural resposes of rats produced by increases in the functional activity of brain 5-hydroxytryptamine and dopamine

Following repeated electroconvulsive shocks (ECS) (once daily for 10 days), rats display enhanced hyperactivity responses to tranylcypromine and l-tryptophan, a procedure which increases brain 5-hydroxytryptamine (5-HT) concentrations, or to the suggested 5-HT agonist quipazine. The enhanced responses last for about 6 days following the last shock. Repeated sub-convulsive shocks did not produce this behavioural enhancement.Administration of indomethacin (2 mg/kg) 25 min before the ECS did not prevent the enhanced 5-HT response suggesting that the enhanced response is not the result of the reported rise in prostaglandins F following ECS.Repeated ECS shortened the time to loss of righting following pentobarbital (50 mg/kg) but did not alter the total sleeping time.Repeated ECS enhances locomotor activity produced by methamphetamine. It also enhances circling produced by methamphetamine and apomorphine in unilateral nigrostriatal lesioned rats, suggesting an enhanced postsynaptic response.No evidence was found for ECS altering the response of striatal adenylate cyclase to dopamine nor for any alteration of striatal cyclic AMP concentration.These data taken with our previous study reinforce the suggestion that electroconvulsive therapy (ECT) produces increased responses to 5-hydroxytryptamine and dopamine receptor stimulation.     

Phospholipase A2 and mediation of the activation of short-circuit current in the rat colonic mucosa

Summary Melittin (0.5–2 g ml^–1) increased the short-circuit current (I_sc) in mucosa-submucosa and mucosa preparations of the rat colon descendens in a dose-dependent manner. In the preparation with the submucosal plexus, quinacrine and indomethacin completely blocked the effect of melittin, indicating activation of phospholipase A₂ and production of prostaglandins induced by the drug. The melittin response was also partially sensitive to the lipoxygenase inhibitor, nordihydroguaiaretic acid. Complete inhibition by tetrodotoxin and atropine gives evidence for the involvement of cholinergic neurons in the mediation of the response induced by melittin. In contrast, in the preparation without the submucosal plexus the effect of melittin was only partially inhibited by quinacrine, indomethacin, or by neuronal blockers, suggesting direct interactions of melittin with the epithelium in addition.The effect of melittin resembles to the action of bradykinin, which is neuronally mediated and quinacrine-sensitive in the mucosa-submucosa preparation, and quinacrine-resistant and not neuronally mediated in the mucosa preparation. In the mucosa-submucosa preparation, the melittin response is even partially sensitive to the bradykinin receptor antagonist [D-Phe⁷]-bradykinin. The results provide evidence for the presence of a quinacrine-sensitive phospholipase A₂ in the preparation with and that without the submucosa. Send offprint requests to: M. Diener at the above address     

Development of tolerance to the antinociceptive effect of metergoline

Metergoline given IP reduced the response to noxious stimulation in the mouse formalin test. Tolerance to this effect developed after a chronic treatment schedule consisting of ten daily injections of 5 mg/kg. Twenty four hours after the last injection a test dose of metergoline (2.5 mg/kg) reduced the licking time in the formalin test by 28% in the chronic metergoline group, compared to 68% reduction in the vehicle-treated animals. In addition, the antinociceptive effect of the 5-hydroxytryptamine releasing compoundp-chloroamphetamine (PCA) was reduced following chronic treatment with metergoline. The reduced effect of PCA may have been caused by down-regulation of 5-HT₂ receptors. However, this finding is also compatible with the contention that metergoline may act as an agonist at postsynaptic serotonergic receptors.     

Effect of sympathomimetic amines on the synaptosomal transport of noradrenaline,dopamine and 5-hydroxytryptamine

The interaction of sympathomimetic amines with the transport of ³H-noradrenaline (³H-NE), ³H-dopamine (³H-DA) and ³H-5-hydroxytriptamine (³H-5-HT) were investigated in rat hypothalamic (³H-NE) and striatal (³H-DA and ³H-5-HT) synaptosomes. Modifications in the phenylethylamine structure led to changes in activity towards biogenic amine uptake and release: (a) the introduction of a β-OH group led to compounds less active in inhibiting uptake and stimulating release of ³H-NE, ³H-DA and ³H-5-HT, with the exception of ³H-NE release which was stimulated more by unlabeled 1-NE than by DA; (b) the introduction of phenolic -OH groups always led to compounds which were stronger uptake inhibitors and releasers of the three biogenic amines; (c) the α-methylation increased the potency towards uptake inhibition and release stimulation, with the exception of ³H-NE release: in fact, the releasing activity of phenylethylamine was suppressed by α-methylation; (d) the introduction of a -Cl group in the para position selectively potentiated the effects on ³H-5-HT uptake and release and generally depressed those on catecholamine transport.     

Neurotoxic effect of maneb in rats as studied by neurochemical and immunohistochemical parameters

Epidemiological investigations document that workers in agriculture, horticulture and people living near areas with frequent use of pesticides have increased risk of developing symptoms of Parkinson's disease. This study investigated the neurotoxic effect of the fungicide maneb by morphological, immunohistochemical and neurochemical methods applying young Sprague–Dawley male rat as the model. Intraperitoneal dosing (7.5, 15 or 30 mg maneb/kg bodyweight/week for 12 weeks) demonstrated dose-related increased manganese concentration in corpus striatum. The striatal concentration of 5-hydroxytryptamine (5-HT) increased in a dose-related manner, as did the 5-HT concentrations in the rest of the brain indicating early sign of neurotoxicity. Striatal acetylcholinesterase activity was not affected. The concentrations of noradrenaline, dopamine, neurotransmitter amino acids and the levels of the proteins -synuclein and synaptophysin in corpus striatum and the rest of the brain were not changed. No histological parameter was affected when studied in corpus striatum and substantia nigra.     

A test of possible cognitive and environmental influences on the mood lowering effect of tryptophan depletion in normal males

In a previous study we found that a tryptophan-deficient amino acid mixture, designed to lower tissue tryptophan and thus brain 5-hydroxytryptamine (5HT) levels, caused a rapid (5 h) lowering of mood in normal males. Because of the importance of this evidence indicating a direct causal connection between low 5HT and low mood, we have now investigated other possible explanations for the mood lowering effect. Research strongly supports the involvement of environmental setting and cognition in the production and experience of emotions. Therefore we investigated how these factors might influence the mood-lowering effects of tryptophan depletion. In an instructional manipulation subjects were either supplied or not supplied with information designed to account for any possible peripheral sensations that might be related to depressive affect. In an environmental manipulation subjects were exposed either to a supportive and comfortable atmosphere (positive environment), or an unrewarding and unstimulating environment (negative environment). In the control group, which received a balanced amino acid mixture, the positive and negative environments had the expected effects on the scores of the Multiple Affect Adjective Checklist, thus indicating the effectiveness of these procedures. In the tryptophan depletion group neither the instructional nor the environmental manipulation had any influence on the mood lowering effect. It may be that tryptophan depletion lowers mood in normal males because low 5HT influences mood directly rather than via cognitive processes. Our data strongly support the idea that 5HT exerts an effect on mood and that low 5HT may, in some patients, be an important factor contributing to the etiology of clinical depression.     

Characterization of 5-hydroxytryptamine (5-HT) receptor subtypes influencing colonic motility in conscious dogs

We examined the effects of exogenous 5-hydroxytryptamine (5-HT) and selective 5-HT receptor agonists and antagonists on proximal, middle and distal colonic motility in conscious fasted dogs with extraluminal force transducers implanted chronically. 5-HT (0.003–0.1 mg/kg i.v.) dose-dependently enhanced motility along the entire length of the colon. The 5-HT (0.03 mg/kg i.v.)-induced response was inhibited by 0.1–1.0 mg/kg i.v. methysergide, a 5-HT_/12 antagonist, at all recording sites and by 0.1–1.0 mg/kg i.v. ketanserin, a 5-HT_2A antagonist, at the middle and distal sites only. At 1 mg/kg i.v., YM060, a 5-HT₃ antagonist, reduced the amplitude of the initial transient high-amplitude contractions induced by 5-HT, but did not affect the tonic contraction induced by 5-HT. At doses up to 3 mg/kg i.v.,2-methoxy-4-amino-5-chlorobenzoic acid 2-(diethylamino) ethyl ester (SDZ205-557), a 5-HT₄ antagonist, and hexamethonium (up to 10 mg/kg i.v.) did not affect 5-HT-induced responses at any recording site. Renzapride, a 5-HT₄ agonist, also stimulated motility along the entire length of the colon at 0.3 mg/kg i.v.. The renzapride-induced response was inhibited by 1 mg/kg i.v. SDZ205-557 or 3 mg/kg i.v. hexamethouium. m-Chlorophenylbiguanide (m-CPBG), a 5-HT₃ agonist, (1 mg/kg i.v.) produced a transient high-amplitude contraction at all recording sites and this contraction was eliminated by pretreatment with 0.03 mg/kg i.v. YM060. The contraction produced by m-CPBG declined rapidly, so the increase in the motility index by m-CPBG was not significant at any recording site. Of the antagonists tested, 0.1–1 mg/kg i.v. methysergide produced a delayed and prolonged contractile response at the middle and distal sites. The onset of the response was delayed about 20 min after application and the response was maintained over the subsequent 60-min observation period. The methysergide (1 mg/kg i.v.)-induced response was inhibited by 3 mg/kg i.v. hexamethonium. The other antagonists, ketanserin, YM060 and SDZ205-557, had no contractile effect at any recording site.These results indicate that exogenous 5-HT stimulates motility along the entire length of the fasted canine colon and that 5-HT-induced responses in the proximal colon are mediated mainly by 5-HT₁, whereas those in the middle and distal colon are mediated by both 5-HT₁ and 5-HT₂ receptors. Renzapride and methysergide also stimulate colonic motility via additional mechanisms. The activation of 5-HT₄ receptors and the blockade of endogenous 5-HT inhibitory regulation via 5-HT₁ receptors may be involved in the action of renzapride and methysergide respectively.     

Reduced effect of morphine in midbrain raphe lesioned rats

The reaction of painful stimulation was studied in rats by means of three different techniques.

Lesions of the midbrain raphe in rats, which reduced the levels of brain 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) decreased the analgesic activity of morphine. In some tests, an increase in sensitivity to painful stimulation was noted in lesioned animals.

The results obtained support the view that a decrease of brain 5-HT concentration antagonizes morphine analgesia. The possible role of the other brain monoamines in the analgesic activity of morphine is also discussed.     

Evidence for two types of 5-hydroxytryptamine receptor on secretomotor neurons of the guinea-pig ileum

Summary Flat sheet preparations of the mucosa plus submucosa from the guinea-pig ileum were placed in Ussing chambers so that short circuit currrent (I _sc), an index of electrolyte movement across the mucosa, could be measured. In these preparations, 5-hydroxytryptamine (5-HT) increasesI _sc indirectly by stimulating both cholinergic and non-cholinergic secretomotor neurons. The 5-HT₃ receptor antagonist, ICS 205–930 (10^–13–10^–5 M), substantially depressed the secretory response due to 5-HT (10^–6 M), but not that produced by direct activation of muscarinic receptors on the mucosal epithelium with carbachol (10^–6 M), or by stimulation of secretomotor neurons with substance P (10^–8 M) or 1,1-dimethyl-4-phenylpiperazinium (10^–5 M). The residual response to 5-HT, after the addition of a maximally effective concentration of ICS 205–930 (10^–6 M), was further reduced by hyoscine (10^–7M). When that part of the 5-HT response attributable to the release of acetylcholine was blocked by hyoscine (10^–7M), ICS 205–930 did not further modify the response to 5-HT. The hyoscine-resistant component was, however, sustantially depressed by tetrodotoxin (3.5 × 10^–7 M). The response remaining after ICS 205–930 and hyoscine was not affected by methysergide (2 × 10^{– 5} M) or cyproheptadine (10^–7 M). We conclude that there are ICS 205–930 sensitive 5-HT receptors on cholinergic secretomotor neurons, and ICS 205–930, methysergide, and cyproheptadine insensitive 5-HT receptors on non-cholinergic secretomotor neurons.     

Effect of central 5-hydroxytryptamine depletion on performance in the free-operant psychophysical procedure: facilitation of switching, but no effect on temporal differentiation of responding

This experiment examined the effect of destroying the ascending 5-hydroxytryptaminergic (5-HTergic) pathways on timing and switching behaviour in the free-operant psychophysical procedure. Rats received injections of 5,7-dihydroxytryptamine into the dorsal and median raphe nuclei or sham lesions. They were trained to press levers for sucrose reinforcement; sessions consisted of fifty 50-s trials in which reinforcers were available on a variable-interval 30-s schedule. In the first 25 s, of each trial, reinforcement was only available for responses on lever A; in the last 25 s, it was available only for responses on lever B. In phase 1 (70 sessions) repetitive switching between the levers was prevented by withdrawal of lever A after the first response on lever B in each trial; in phase 2 (40 sessions) this constraint on switching was removed; in phase 3 (40 sessions) the constraint was reinstated. Data were collected from probe trials (four per session) in which no reinforcers were delivered, during the last ten sessions of each phase. In all phases, both groups showed declining response rates on lever A and increasing response rates on lever B as a function of time from the onset of the trial. Response rate on lever B, expressed as percentage of overall response rate, could be described by a two-parameter logistic function. Removal of the constraint on switching reduced the slope of the function without changing the indifference point (time corresponding to 50% responding on lever B). The parameters of the timing function did not differ between the groups in any of the phases. However, the lesioned group showed a greater enhancement of switching rate during phase 2 than the control group. The levels of 5-HT and 5-hydroxyindoleacetic acid were reduced in the brains of the lesioned rats, but the levels of noradrenaline and dopamine were not altered. The results provide further evidence for the involvement of the ascending 5-HTergic pathways in switching between response alternatives, but cast doubt on our previous suggestion that the effects of 5-HT depletion on temporal differentiation of behaviour are mediated by facilitated switching. Received: 12 July 1998/Final version: 9 October 1998