RESUMO
BACKGROUND AND PURPOSE: Hypothalamic neuropeptides centrally modulate sexual arousal. However, the role of neuropeptides in peripheral arousal has been ignored. Vascular and non-vascular smooth muscle relaxation in the vagina is important for female sexual arousal. To date, in vitro studies have focused on vaginal strips with no studies on vaginal arteries. The aim of this study was to compare the effects of sexual hypothalamic neuropeptides on rabbit vaginal wall strips and arteries. EXPERIMENTAL APPROACH: Tissue bath and wire myography techniques were used to measure isometric tension from strips and arteries, respectively. KEY RESULTS: Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) relaxed both preparations, effects that were only antagonized by the VIP/PACAP antagonist VIP6-28 (10 nM) and the PAC(1) antagonist PACAP 6-38 (1 microM). The melanocortin agonist alpha-melanocortin-stimulating hormone (1 microM), but not bremelanotide (1 microM), also relaxed both preparations. Oxytocin and vasopressin contracted vaginal preparations, which could be antagonized by the V(1A) antagonist SR 49059. Neuropeptide Y (NPY) and the NPY Y(1) agonist Leu(31), Pro(34) NPY only contracted arteries, which was antagonized by the NPY Y(1) receptor antagonist BIBP 3226. Melanin-concentrating hormone (MCH; 1 microM) contracted arteries. CONCLUSION AND IMPLICATIONS: Hypothalamic neuropeptides can exert contractile and relaxant effects on vaginal strips and arteries. NPY Y(1), V(1A), MCH(1) antagonists as well as VIP/PAC(1) agonists may have therapeutic potential in both central and peripheral female sexual arousal. Differences in effect of neuropeptides between preparations raise the question of which preparation is important for female sexual arousal.
Assuntos
Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Neuropeptídeos/farmacologia , Vagina/anatomia & histologia , Animais , Feminino , Modelos Moleculares , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Músculo Liso Vascular/fisiologia , Neuropeptídeo Y/farmacologia , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/farmacologia , Conformação Proteica , Coelhos , Peptídeo Intestinal Vasoativo/farmacologiaRESUMO
BACKGROUND AND PURPOSE: Maintained penile erection depends on the absence of alpha-adrenoceptor (alpha-AR) activation and so can be facilitated by alpha-blockers. This study seeks the alpha(1)-AR subtypes involved in order to inform the pro-erectile consequences of subtype selective blockade. EXPERIMENTAL APPROACH: Wire myography was used with dorsal (nutritional supply) and cavernous (erectile inflow) penile arteries; standard alpha-AR-selective agonists and antagonists were employed to classify responses. KEY RESULTS: In both penile arteries noradrenaline (NA) and phenylephrine (PE, alpha(1)-AR agonist) caused concentration-dependent contractions. Sensitivity to NA was increased by NA uptake blockers, cocaine (3 microM) and corticosterone (30 microM). PE responses were antagonised by phentolamine (non-selective alpha-AR: dorsal pK(B) 8.00, cavernous 8.33), prazosin (non-subtype-selective alpha(1)-AR: dorsal 8.60, cavernous 8.41) and RS100329 (alpha(1A)-AR selective: dorsal 9.03, cavernous 8.80) but not by BMY7378 (alpha(1D)-AR selective: no effect at 1-100 nM) or Rec15/2615 (alpha(1B)-AR selective: no effect at 1-100 nM). Schild analysis was straightforward in cavernous artery, indicating that PE activates only alpha(1A)-AR. In dorsal artery Schild slopes were low, though alpha(1A)-AR was still indicated. Analysis using UK 14,304 and rauwolscine indicated an alpha(2)-AR component in dorsal artery that may account for low slopes to alpha(1)-AR antagonists. CONCLUSIONS AND IMPLICATIONS: Penile arteries have a predominant, functional alpha(1A)-AR population with little evidence of other alpha(1)-AR subtypes. Dorsal arteries (nutritional supply) also have alpha(2)-ARs. Thus, alpha-AR blockers with affinity for alpha(1A)-AR or alpha(2)-AR would potentially have pro-erectile properties; the combination of these perhaps being most effective. This should inform the design of drugs to assist/avoid penile erection.
Assuntos
Artérias/efeitos dos fármacos , Pênis/irrigação sanguínea , Receptores Adrenérgicos alfa 1/fisiologia , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Artérias/fisiologia , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Fenilefrina , CoelhosRESUMO
BACKGROUND AND PURPOSE: Ghrelin increases growth hormone secretion, gastric acid secretion, gastric motility and hunger but decreases glucose-dependent insulin secretion and insulin sensitivity in humans. Antagonizing the ghrelin receptor has potential as a therapeutic approach in the treatment of obesity and type 2 diabetes. Therefore, the aim was to pharmacologically characterize the novel small-molecule antagonist PF-05190457 and assess translational pharmacology ex vivo. EXPERIMENTAL APPROACH: Radioligand binding in filter and scintillation proximity assay formats were used to evaluate affinity, and europium-labelled GTP to assess functional activity. Rat vagal afferent firing and calcium imaging in dispersed islets were used as native tissues underlying food intake and insulin secretion respectively. KEY RESULTS: PF-05190457 was a potent and selective inverse agonist on constitutively active ghrelin receptors and acted as a competitive antagonist of ghrelin action, with a human Kd of 3 nM requiring 4 h to achieve equilibrium. Potency of PF-05190457 was similar across different species. PF-05190457 increased intracellular calcium within dispersed islets and increased vagal afferent firing in a concentration-dependent manner with similar potency but was threefold less potent as compared with the in vitro Ki in recombinant overexpressing cells. The effect of PF-05190457 on rodent islets was comparable with glibenclamide, but glucose-dependent and additive with the insulin secretagogue glucagon-like peptide-1. CONCLUSIONS AND IMPLICATIONS: Together, these data provide the pharmacological in vitro and ex vivo characterization of the first ghrelin receptor inverse agonist, which has advanced into clinical trials to evaluate the therapeutic potential of blocking ghrelin receptors in obesity and type 2 diabetes.
Assuntos
Azetidinas/farmacologia , Agonismo Inverso de Drogas , Glucose/metabolismo , Insulina/metabolismo , Receptores de Grelina/antagonistas & inibidores , Compostos de Espiro/farmacologia , Nervo Vago/efeitos dos fármacos , Animais , Azetidinas/química , Cálcio/metabolismo , Relação Dose-Resposta a Droga , Células HEK293 , Humanos , Secreção de Insulina , Masculino , Ratos , Ratos Sprague-Dawley , Compostos de Espiro/química , Relação Estrutura-Atividade , Nervo Vago/metabolismo , Nervo Vago/fisiologiaRESUMO
While nicotine is known to act at neuronal nicotinic acetylcholine receptors (nAChRs) to facilitate neurotransmitter release, the mechanisms underlying this action are poorly understood. Some of its effects are known to be mediated by presynaptic receptors. In the mouse vas deferens nicotine (10-30 microM) transiently increased the force of neurogenic contraction by 135+/-25%, increased the amplitude of excitatory junction potentials by 74+/-6% and increased the frequency of spontaneous excitatory junction potentials in four out of six preparations. Confocal microscopy and the calcium indicator Oregon Green 488 BAPTA-1 dextran were used to measure calcium concentration changes in the nerve terminals. Nicotine did not affect the action potential-evoked calcium transient but instead triggered small, random fluctuations ("calcium spikes") in intra-varicosity calcium concentrations at an average frequency of 0.09+/-0.02 Hz. These were insensitive to tetrodotoxin at a concentration that blocked action-potential evoked calcium transients (300 nM). They were abolished by the nAChR blocker hexamethonium (100 microM) and by both ryanodine (100 microM) and caffeine (3 mM), agents that modify calcium release from intracellular stores. We propose a novel mechanism whereby nicotine's action at nAChRs triggers calcium-induced calcium release from a ryanodine-sensitive calcium store in nerve terminals. This primes neurotransmitter release mechanisms and enhances both spontaneous and action potential-evoked neurotransmitter release.
Assuntos
Sinalização do Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Líquido Intracelular/efeitos dos fármacos , Junção Neuromuscular/efeitos dos fármacos , Nicotina/farmacologia , Terminações Pré-Sinápticas/efeitos dos fármacos , Receptores Nicotínicos/efeitos dos fármacos , Antagonistas Adrenérgicos alfa/farmacologia , Animais , Cafeína/farmacologia , Canais de Cálcio/efeitos dos fármacos , Canais de Cálcio/metabolismo , Sinalização do Cálcio/fisiologia , Estimulação Elétrica , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Corantes Fluorescentes/farmacocinética , Hexametônio/farmacologia , Líquido Intracelular/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/inervação , Músculo Liso/fisiologia , Junção Neuromuscular/metabolismo , Junção Neuromuscular/ultraestrutura , Antagonistas Nicotínicos/farmacologia , Compostos Orgânicos , Prazosina/farmacologia , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Receptores Nicotínicos/metabolismo , Rianodina/farmacologia , Fibras Simpáticas Pós-Ganglionares/citologia , Fibras Simpáticas Pós-Ganglionares/efeitos dos fármacos , Fibras Simpáticas Pós-Ganglionares/metabolismo , Tetrodotoxina/farmacologia , Ducto Deferente/efeitos dos fármacos , Ducto Deferente/inervação , Ducto Deferente/fisiologiaRESUMO
PURPOSE: In nongenital arteries a sex difference has been postulated in the dominant endothelium-derived relaxant factor(s), eg nitric oxide, prostacyclin or endothelial-derived hyperpolarizing factor. Knowledge of endothelium-derived relaxant factor mechanisms in genital tissues could influence the development of novel treatments for sexual dysfunction. We compared nitric oxide and endothelial-derived hyperpolarizing factor contributions to acetylcholine induced relaxation in the genital arteries of the 2 sexes. MATERIALS AND METHODS: Male dorsal and cavernous penile arteries, and female extravaginal and intravaginal arteries from New Zealand White rabbits were studied. Acetylcholine concentration-vasodilator response curves were constructed in the presence of the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester, K(+) channel blockers (apamin and charybdotoxin) or a combination. Indomethacin was present throughout to exclude prostacyclins. RESULTS: Extravaginal artery relaxation was predominantly endothelial-derived hyperpolarizing factor induced. Apamin plus charybdotoxin decreased maximal relaxations from a mean +/- SEM of 77% +/- 4% to 23% +/- 3% in 6 preparations (p <0.01). However, nitric oxide and endothelial-derived hyperpolarizing factor contributed to overall function. Dorsal artery relaxation was largely nitric oxide induced. Nomega-nitro-L-arginine methyl ester decreased maximal relaxations from 90% +/- 3% to 41% +/- 9% (p <0.001) with no endothelial-derived hyperpolarizing factor involvement (p >0.05). In cavernous and intravaginal arteries nitric oxide and endothelial-derived hyperpolarizing factor contributed to acetylcholine induced relaxation, while nitric oxide predominated. Blocking nitric oxide synthase or K(+) channels indicated that myogenic tone and constitutive activity of endothelium-derived relaxant factors were present. Vasodilator nerve mediated responses were influenced by each with the former more effective. CONCLUSIONS: Vaginal inflow arteries showed a dominance of endothelial-derived hyperpolarizing factor, contrasting with nitric oxide in penile arteries. Penile arteries followed the trend that endothelial-derived hyperpolarizing factor involvement increased with decreasing vessel caliber, while the reverse was demonstrated in female arteries.
Assuntos
Fatores Biológicos/fisiologia , Endotélio Vascular/fisiologia , Óxido Nítrico/fisiologia , Pênis/irrigação sanguínea , Pênis/fisiologia , Vagina/irrigação sanguínea , Vagina/fisiologia , Animais , Feminino , Masculino , Relaxamento Muscular , Coelhos , Caracteres Sexuais , Resistência VascularRESUMO
The course of twin pregnancy is most amenable to positive intervention during the antepartal period. However, once a laboring woman arrives on the labor floor, there is still opportunity to influence outcome. The management of multiple gestation is best accomplished through use of a multidisciplinary team. Intrapartal events are handled with greater confidence when each member of the team is aware not only of his or her specific role, but also of all the tasks that need to be accomplished. Rehearsal prior to the delivery and communication during the delivery are essential. Each team member can positively affect the outcome in twin delivery through the use of unique skills. The preceding review of the literature regarding multiple gestation during labor portrays the dilemmas and controversies surrounding management. It is incumbent on all maternity staff to become acquainted with the issues and acquire the skills necessary to render safe care. Every complication of labor and delivery, including preterm labor, uterine dysfunction, abnormal presentation, prolapse of the umbilical cord, premature separation of the placenta, and immediate postpartum hemorrhage, occurs with greater frequency with multiple gestation. Therefore, the conduct of labor and delivery with twins is an excellent challenge to the skills of the team that provides care for the patient and her fetuses.
Assuntos
Parto Obstétrico , Gravidez Múltipla , Gêmeos , Feminino , Humanos , Recém-Nascido , Admissão do Paciente , Equipe de Assistência ao Paciente , Gravidez , Cuidado Pré-NatalRESUMO
In 1969, Paton and Vizi described the inhibitory actions of noradrenaline on acetylcholine release from the innervation of the guinea-pig ileum longitudinal muscle. They concluded "that acetylcholine output by the nervous networks of the longitudinal strip is under the normal control of the sympathetic by a species of presynaptic inhibition mediated by <==> receptors". This work was carried out in the Pharmacology Department at Oxford University. Clearly, a period in the 'Dreaming Spires' of Oxford sufficiently inspired Sylvester to take up a life long career in scientific research. He has published more than 300 papers on a wide range of topics but clearly has a strong interest in neurotransmitter release mechanisms and recently, non-synaptic interactions between neurons. It seems fitting therefore to write a brief review on the continuing studies on neurotransmitter release mechanisms in sympathetic neurons in a volume honoring the now distinguished Professor Vizi.
Assuntos
Neurônios/fisiologia , Neurotransmissores/fisiologia , Sistema Nervoso Simpático/fisiologia , Animais , Potenciais Pós-Sinápticos Excitadores , Masculino , Neurotransmissores/metabolismo , Sistema Nervoso Simpático/citologia , Sistema Nervoso Simpático/metabolismo , Ducto Deferente/inervaçãoRESUMO
1. Action potential-evoked Ca(2+) transients in postganglionic sympathetic axon bundles in mouse vas deferens have been characterized using confocal microscopy and Ca(2+) imaging. 2. Axonal Ca(2+) transients were tetrodotoxin sensitive. The amplitude depended on both the frequency of stimulation and the number of stimuli in a train. 3. Removal of extracellular Ca(2+) abolished the Ca(2+) transient. Cd(2+)(100 microM) inhibited the Ca(2+) transient by 78 +/- 10 %. The N-type Ca(2+) channel blocker omega-conotoxin GVIA (0.1 microM) reduced the amplitude by -35 +/-4 %, whereas nifedipine (10 microM; L-type) and omega-conotoxin MVIIC (0.1 microM; P/Q type) were ineffective. 4. Caffeine (10 mM), ryanodine (10 microM), cyclopiazonic acid (30 microM) or CCCP (10 microM) had no detectable effects. 5. Blockade of large and small conductance Ca(2+)-dependent K+ channels with iberiotoxin (0.1 microM) and apamin (1 microM), respectively, or Ca(2+)-dependent Cl(-) channels by niflumic acid (100 microM) did not alter Ca(2+) transients. 6. In contrast, the non-specific K+ channel blockers tetraethylammonium (10 mM) and 4-aminopyridine (10 mM) markedly increased the amplitude of the Ca(2+) transient. Blockade of delayed rectifiers and A-like K+ channels, by tityustoxin-K (alpha) (0.1 microM) and pandinustoxin-K (alpha) (10 nM), respectively, also increased the Ca(2+) transient amplitude. 7. Thus, Ca(2+) transients are evoked by Na(+)-dependent action potentials in axons. These transients originate mainly from Ca(2+) entry through voltage-dependent Ca(2+) channels (80 % Cd(2+) sensitive of which 40 % was attributable to N-type). Twenty per cent of the Ca(2+) transient was not due to Ca(2+) entry through voltage-gated Ca(2+) channels. Intracellular stores and mitochondria were not involved in the generation of the transient. Ca(2+) transients are modulated by A-like K+ channels and delayed rectifiers (possibly K(V)1.2) but not by Ca(2+)-activated ion channels.
Assuntos
Axônios/fisiologia , Cálcio/fisiologia , Gânglios/fisiologia , Sistema Nervoso Simpático/fisiologia , Potenciais de Ação/fisiologia , Animais , Canais de Cálcio/metabolismo , Canais de Cloreto/antagonistas & inibidores , Estimulação Elétrica/métodos , Eletrofisiologia , Membranas Intracelulares/metabolismo , Ativação do Canal Iônico , Ionomicina/farmacologia , Ionóforos/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Bloqueadores dos Canais de Potássio/farmacologia , Canais de Potássio/fisiologia , Tetrodotoxina/farmacologia , Fatores de TempoRESUMO
1. Development of the pacemaker system in the small intestine depends upon signalling via tyrosine kinase (Kit) receptors. The downstream pathways initiated by Kit in interstitial cells of Cajal (ICC) have not been investigated. Wortmannin and 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY 294002), inhibitors of phosphatidylinositol 3'-kinase (PI3-kinase), were used to test the involvement of this pathway in the development and maintenance of ICC and electrical rhythmicity in the murine small intestine. 2. ICC and electrical slow waves were present in the murine jejunum at birth. ICC and electrical rhythmicity continued to develop in neonates such that adult activity was recorded after 1 week. Development of ICC and rhythmicity were maintained in organ culture. 3. Wortmannin or LY 294002 inhibited the development of slow waves and blocked rhythmicity within 2-4 days. Loss of slow waves was preceded by disappearance of Kit-positive cells from the myenteric (IC-MY) and deep muscular plexus (IC-DMP) regions. Wortmannin or LY 294002 had no acute effect on slow waves. 4. Muscles from older animals (day 10-day 30) developed resistance to wortmannin treatment, but when the exposure to wortmannin was increased to 35 days, damage to ICC networks and electrical dysrhythmias were observed. 5. PI3-kinase appears to be a critical downstream signalling element linking Kit receptors to ICC development and maintenance of phenotype. ICC are more sensitive to Kit or PI3-kinase blockade at birth, but the importance of the PI3-kinase signalling in the maintenance of ICC persists into adulthood. Interference with PI3-kinase signalling in immature or adult animals could result in disruption of ICC and gastrointestinal dysrhythmias.