Serotonin contracts the rat mesenteric artery by inhibiting 4-aminopyridine-sensitive Kv channels via the 5-HT2A receptor and Src tyrosine kinase

Serotonin (5-hydroxytryptamine (5-HT)) is a neurotransmitter that regulates a variety of functions in the nervous, gastrointestinal and cardiovascular systems. Despite such importance, 5-HT signaling pathways are not entirely clear. We demonstrated previously that 4-aminopyridine (4-AP)-sensitive voltage-gated K+ (Kv) channels determine the resting membrane potential of arterial smooth muscle cells and that the Kv channels are inhibited by 5-HT, which depolarizes the membranes. Therefore, we hypothesized that 5-HT contracts arteries by inhibiting Kv channels. Here we studied 5-HT signaling and the detailed role of Kv currents in rat mesenteric arteries using patch-clamp and isometric tension measurements. Our data showed that inhibiting 4-AP-sensitive Kv channels contracted arterial rings, whereas inhibiting Ca2+-activated K+, inward rectifier K+ and ATP-sensitive K+ channels had little effect on arterial contraction, indicating a central role of Kv channels in the regulation of resting arterial tone. 5-HT-induced arterial contraction decreased significantly in the presence of high KCl or the voltage-gated Ca2+ channel (VGCC) inhibitor nifedipine, indicating that membrane depolarization and the consequent activation of VGCCs mediate the 5-HT-induced vasoconstriction. The effects of 5-HT on Kv currents and arterial contraction were markedly prevented by the 5-HT2A receptor antagonists ketanserin and spiperone. Consistently, alpha-methyl 5-HT, a 5-HT2 receptor agonist, mimicked the 5-HT action on Kv channels. Pretreatment with a Src tyrosine kinase inhibitor, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, prevented both the 5-HT-mediated vasoconstriction and Kv current inhibition. Our data suggest that 4-AP-sensitive Kv channels are the primary regulator of the resting tone in rat mesenteric arteries. 5-HT constricts the arteries by inhibiting Kv channels via the 5-HT2A receptor and Src tyrosine kinase pathway.

Estudio comparativo de la cicatrización en piel de conejo entre sucralfato y tartrato de ketanserin / Comparative study of wound healing in rabbit skin between sucralfate and ketanserin tartrate

Al sanar una herida se activa un proceso de reepitelización para generar una epidermis. Este proceso de cicatrización debe ocurrir rápido y efectivamente para prevenir los ataques provenientes del medio ambiente. Existe una gran variedad de cremas regeneradoras, dentro de las cuales se encuentran sucralfato (Cicalfate ®), de utilidad en medicina humana, y tartrato de ketanserin (Vulketan ®), utilizado como tratamiento convencional en medicina veterinaria. El objetivo general de este estudio fue comparar el grado de cicatrización y reacción inflamatorio entre sucralfato y tartrato de ketanserin en heridas quirúrgicas en piel del área superior de la pared torácica de conejos (Oryctologus cuniculis), los cuales se utilizaron como modelos animales para medir parámetros cicatriciales que son complicados de evaluar en forma rutinaria en estudios clínicos en el ser humano. Se emplearon 15 conejos machos. Sucralfato y tartrato de ketonserin fueron aplicados simultáneamente en 10 animales. Sucralfato se aplicó en el lado derecho y tartrato de ketanserin, en el lado izquierdo. Cinco conejos fueron tratados con suero fisiológico como grupo control. La evaluación se realizó mediante la Escala de Vancouver durante 19 días. Los resultados revelan que en el caso de ambas cremas regeneradoras no existe diferencia significativa en torno a los signos de vascularidad. Con respecto o los signos de plegabilidad, altura, pigmentación y dolor, sí existe diferencia significativa (P < 0,05) contribuyendo mejor sucralfato, por lo que se concluye que éste presento ventaja comparativa con respecto o tartrato de ketanserin al momento de realizar un tratamiento tópico para heridas en piel.

In healing o process of reepithelialization to generate epidermis start. This process must take place quickly and effectively to prevent attacks from the environment. There is a great variety of regenerative creams available such as sucralfate (Ciclalfate®) in human medicine and ketaserin tartrate (Vulketan®) used as a conventional treatment in veterinary medicine. The main objective of this study was to compare the healing degree and inflammatory reaction between sucralfate and ketanserin tartrate in surgical wounds on skin of the upper thoracic wall of rabbits (Oryctolagus cuniculis), which were used as animal models for measuring cicatricial parameters that are complicated to evaluate routinely in clinical trials in humans. Fifteen male rabbits were used Sucralfate and ketanserin tartrate were used simultaneously in 10 animals. Sucralfate was used on right side and ketanserin trartrate on the left side. Five rabbits were treated with physiological saline solution (control group). Wound evaluation were compared with the Scale of Vancouver during 19 days. The results showed that both regenerative creams had no significant differences in vascularity. In relation to pliability height, pigmentation and pain there was a significant difference (P < 0,05) between the two products in favor of sucralfate in the topical treatment of skin injuries.

Behavioral and neuropharmacological evidence that serotonin crosses the blood-brain barrier in Coturnix japonica (Galliformes; Aves)

This study was carried out aiming to reach behavioral and neuropharmacological evidence of the permeability of the blood-brain barrier (BBB) to serotonin systemically administered in quails. Serotonin injected by a parenteral route (250-1000 æg.kg-1, sc) elicited a sequence of behavioral events concerned with a sleeping-like state. Sleeping-like behaviors began with feather bristling, rapid oral movements, blinking and finally crouching and closure of the eyes. Previous administration of 5-HT2C antagonist, LY53857 (3 mg.kg-1, sc) reduced the episodes of feather bristling and rapid oral movements significantly but without altering the frequency of blinking and closure of the eyes. Treatment with the 5-HT2A/2C antagonist, ketanserin (3 mg.kg-1, sc) did not affect any of the responses evoked by the serotonin. Quipazine (5 mg.kg-1, sc) a 5-HT2A/2C/3 agonist induced intense hypomotility, long periods of yawning-like and sleeping-like states. Previous ketanserin suppressed gaping responses and reduced hypomotility, rapid oral movements and bristling but was ineffective for remaining responses induced by quipazine. Results showed that unlike mammals, serotonin permeates the BBB and activates hypnogenic mechanisms in quails. Studies using serotoninergic agonist and antagonists have disclosed that among the actions of the serotonin, feather bristling, rapid oral movements and yawning-like state originated from activation of 5-HT2 receptors while blinking and closure of the eyes possibly require other subtypes of receptors.

Este estudo foi desenvolvido objetivando ampliar as evidências comportamentais e neurofarmacológicas da permeabilidade da barreira hematoencefálica (BHE) à serotonina administrada sistemicamente em codornas. A serotonina injetada por via parenteral (250-1000 æg.kg-1, sc) produziu uma seqüência de eventos relacionados com um estado semelhante ao sono. Comportamentos semelhantes ao sono começaram com o eriçamento das penas, movimentos orais rápidos, piscadelas e finalmente agachamento e fechamento dos olhos. A administração prévia do antagonista do receptor 5-HT2C, LY53857 (3 mg.kg-1, sc) reduziu significativamente os episódios de eriçamento das penas e movimentos orais rápidos, mas não alterou a freqüência de piscadelas e fechamento dos olhos. Tratamento com o antagonista do receptor 5-HT2A/2C, quetanserina (3 mg.kg-1, sc) não afetou nenhuma das respostas evocadas pela serotonina. A quipazina (5 mg.kg-1, sc), um agonista dos receptores 5-HT2A/2C/3, induziu intensa hipomotilidade e longos períodos de comportamentos semelhantes ao bocejo e ao sono. O tratamento prévio com quetanserina suprimiu as reações de bocejo e reduziu a hipomotilidade, os movimentos orais rápidos e as piscadelas, mas foi sem efeito para as demais respostas induzidas pela quipazina. Os resultados mostraram que, diferentemente dos mamíferos, a serotonina atravessa a BHE e ativa mecanismos hipnogênicos em codornas. Estudos com agonistas serotoninérgicos e antagonistas revelaram que, entre as ações da serotonina, o eriçamento das penas, os movimentos orais rápidos e o comportamento semelhante ao bocejo foram originados pela ativação de receptores 5-HT2, enquanto o piscar e o fechamento dos olhos possivelmente requereu outros subtipos de receptores.

Maternal aggression in Wistar rats: effect of 5-HT2A/2C receptor agonist and antagonist microinjected into the dorsal periaqueductal gray matter and medial septum

The objective of the present study was to assess the role of the 5-HT2A/2C receptor at two specific brain sites, i.e., the dorsal periaqueductal gray matter (DPAG) and the medial septal (MS) area, in maternal aggressive behavior after the microinjection of either a 5-HT2A/2C receptor agonist or antagonist. Female Wistar rats were microinjected on the 7th postpartum day with the selective agonist alpha-methyl-5-hydroxytryptamine maleate (5-HT2A/2C) or the antagonist 5-HT2A/2C, ketanserin. The agonist was injected into the DPAG at 0.2 (N = 9), 0.5 (N = 10), and 1.0 æg/0.2 æl (N = 9), and the antagonist was injected at 1.0 æg/0.2 æl (N = 9). The agonist was injected into the medial septal area (MS) at 0.2 (N = 9), 0.5 (N = 7), and 1.0 æg/0.2 æl (N = 6) and the antagonist was injected at 1.0 æg/0.2 æl (N = 5). For the control, saline was injected into the DPAG (N = 7) and the MS (N = 12). Both areas are related to aggressive behavior and contain a high density of 5-HT receptors. Non-aggressive behaviors such as horizontal locomotion (walking) and social investigation and aggressive behaviors such as lateral threat (aggressive posture), attacks (frontal and lateral), and biting the intruder were analyzed when a male intruder was placed into the female resident's cage. For each brain area studied, the frequency of the behaviors was compared among the various treatments by analysis of variance. The results showed a decrease in maternal aggressive behavior (number of bites directed at the intruder) after microinjection of the agonist at 0.2 and 1.0 æg/0.2 æl (1.6 ± 0.7 and 0.9 ± 0.3) into the DPAG compared to the saline group (5.5 ± 1.1). There was no dose-response relationship with the agonist. The present findings suggest that the 5-HT2A/2C receptor agonist has an inhibitory effect on maternal aggressive behavior when microinjected into the DPAG and no effect when microinjected into the MS. Ketanserin (1.0 æg/0.2 æl) decreased locomotion when microinjected into the DPAG and MS, but did not affect aggressive behavior. We interpret these findings as evidence for a specific role of 5-HT2A/2C receptors in the DPAG in the inhibition of female aggressive behavior, dissociated from those on motor activity.

Investigation of central mechanism of insulin induced hypoglycemic convulsions in mice.

Insulin produces seizures in healthy and diabetic animals. Amongst suggested mechanisms, the role of neuromodulators and neurotransmitters is not clear. The present study explores the mechanisms involved in insulin-induced convulsions. Convulsions were induced in Swiss male albino mice with graded doses of insulin. Blood sugar levels were measured prior to and after the first convulsion. Drugs like 5-HTP (5-HT precursor), pCPA (5-HT depletor), ondansetron (5-HT3 antagonist), ketanserin (5-HT, antagonist), ketamine (NMDA antagonist), 1-dopa (dopamine precursor) and reserpine (amine depletor) were studied for interaction with convulsive behaviour induced by insulin. Insulin in 2 IU/kg dose did not produce convulsions while 4 and 8 IU/kg doses produced convulsions in 50% and 100% of animals respectively. 5-HTP, ondansetron, ketanserin, ketamine and l-dopa significantly protected/inhibited animals from convulsions at all studied doses of insulin. On the contrary, pCPA and reserpine potentiated insulin induced convulsions. Insulin caused mortality in 40 and 100% animals with 4 and 8 IU/kg doses respectively. pCPA and reserpine treatments caused mortality at all doses of insulin, while other drugs did not influence insulin induced mortality. Blood sugar levels were reduced in all groups irrespective of the presence or absence of convulsions. A definitive link of serotonergic, dopaminergic and excitatory amino acid pathways in mediating insulin-induced hypoglycemic convulsions is suggested.

Behavioral effects of intra-amygdala injections of GABA and 5-HT acting drugs in the elevated plus-maze

The effect of drugs bilaterally injected into the basolateral/medial nuclei of the amygdala on the behavior of male Wistar rats (300-330 g) in the elevated plus-maze was measured. The benzodiazepine agonist midazolam (MDZ, 20 and 40 nmol, 0.2 µl; N = 8-14) significantly increased open-arm exploration (percent open-arm entries; control = 20.27 + or - 3.71; 40 nmol MDZ = 42.63 + or - 7.16), having thus an anxiolytic effect. On the contrary, the non-selective 5-HT2 antagonist ketanserin (KET, 1 and 10 nmol, 0.2 µl; N = 8-11) had an anxiogenic effect (percent open-arm entries: control = 35.61 + or - 6.41; 10 nmol KET = 18.65 + or - 3.89). The 5-HT1A full agonist 8-OH-DPAT (2, 4 and 8 nmol, 0.2 µl; N = 9.12) did not significantly achange rat behavior in the plus-maze. While the present anxiolytic effect of midazolam agrees with results reported by others using punished behavior, the effect of the serotonergic drugs does not. Therefore, the effect of 5-HT acting drugs injected into the amygdala may be determined by the type of experimental model of anxiety used

Influence of atropine and ketanserin on canine jejunal secretory and contractile effects of serotonin

A serotonina é um importante mediador da secreçao hidroeletrolítica e da motilidade intestinais. Diversas substâncias têm sido estudadas no sentido de controlar os efeitos hipersecretórios e motores, com conseqüênte diarréia severa, induzidos pela hiperserotoninemia. Este trabalho investiga a influência da atropina e da quetaserina sobre a secreçao hidroeletrolítia e a motilidade jejunais induzidas pela serotonina, en alças de Thiry-Vella de cao. Ambas as substâncias reduziram a secreçao de água, sódio, potássio e cloreto, bem como diminuíram a atividade motora de segmento jejunal isolado. Esses resultados sugerem uma possível açao benéfica da atropina e da quetanserina no tratamento de efeitos prejudiciais da serotonina sobre o intestino, como por exemplo a diarréia

Serotonergic modulation of footshock induced aggression in paired rats.

Footshock induced aggression (FIA) was induced in paired rats and three paradigms of aggressive behaviour were recorded, namely, latency to fight (LF), total period of physical contact (TPP) and cumulative aggression scores (CAS). The effects of increasing or decreasing central serotonergic activity, by using a number of pharmacological agents with well defined effects on rat brain serotonin, were investigated on FIA and on FIA augmented by apomorphine, a dopamine receptor agonist. The results show that centrally administered serotonin, the serotonin precursor, 5-hydroxytryptophan administered with clorgyline, a selective MAO A inhibitor, quipazine, a serotonin receptor agonist, and fluoxetine, a selective inhibitor of neuronal re-uptake of serotonin, attenuated all paradigms of FIA and apomorphine induced potentiation of FIA. On the contrary, the other re-uptake inhibitor used, citalopram, appeared to have a dual effect and decreased LF and CAS, while increasing TPP. The serotonin synthesis inhibitor, p-chlorophenylalanine and the selective serotonin receptor (5-HT2) antagonist, ketanserin, augmented all paradigms of FIA per se and apomorphine induced augmentation of FIA. However, the other serotonin receptor antagonist used, metergoline, which blocks both 5-HT1 and 5-HT2 receptor subtypes, attenuated FIA per se but decreased only CAS in apomorphine induced increase in FIA. The data confirm the inhibitory effect of the central serotonergic system on aggressive behaviour and the inverse relationship existing between it and the central dopaminergic system in the modulation of FIA, as has also been confirmed in earlier biochemical investigations from this laboratory. The data has been discussed in the light of existing knowledge on serotonin receptor subtypes and the presence of modulatory serotonergic heteroreceptors on central dopaminergic neurones.

Effect of monoamine antagonists on pretectal evoked analgesia in rat.

Mild and brief electrical stimulation of sites in the pretectal nucleus of rat produced analgesia (SPA) of long duration without significant aversion. Intracerebroventricular (icv) administration of 5-HT receptor antagonists methysergide (50 micrograms) and ketanserin (50 micrograms) and the dopaminergic antagonist haloperidol (50 micrograms) had no significant effect on pretectal SPA, but alpha and beta adrenoceptor antagonists phenoxybenzamine (50 micrograms) and sotalol (50 micrograms) on icv injection significantly antagonised the pretectal SPA. The results suggest that pretectal SPA involves activation of central adrenoceptors.