RESUMO
BACKGROUND: Cerebellar ataxias are the result of diverse disease processes that can be genetic or acquired. Establishing a diagnosis requires a methodical approach with expert clinical evaluation and investigations. We describe the causes of ataxia in 1500 patients with cerebellar ataxia. METHODS: All patients were referred to the Sheffield Ataxia Centre, UK, and underwent extensive investigations, including, where appropriate genetic testing using next-generation sequencing (NGS). Patients were followed up on a 6-monthly basis for reassessment and further investigations if indicated. RESULTS: A total of 1500 patients were assessed over 20â years. Twenty per cent had a family history, the remaining having sporadic ataxia. The commonest cause of sporadic ataxia was gluten ataxia (25%). A genetic cause was identified in 156 (13%) of sporadic cases with other causes being alcohol excess (12%) and cerebellar variant of multiple system atrophy (11%). Using NGS, positive results were obtained in 32% of 146 patients tested. The commonest ataxia identified was EA2. A genetic diagnosis was achieved in 57% of all familial ataxias. The commonest genetic ataxias were Friedreich's ataxia (22%), SCA6 (14%), EA2 (13%), SPG7 (10%) and mitochondrial disease (10%). The diagnostic yield following attendance at the Sheffield Ataxia Centre was 63%. CONCLUSIONS: Immune-mediated ataxias are common. Advances in genetic testing have significantly improved the diagnostic yield of patients suspected of having a genetic ataxia. Making a diagnosis of the cause of ataxia is essential due to potential therapeutic interventions for immune and some genetic ataxias.
Assuntos
Ataxia Cerebelar/etiologia , Adulto , Encéfalo/diagnóstico por imagem , Ataxia Cerebelar/diagnóstico , Ataxia Cerebelar/genética , Diagnóstico Diferencial , Inglaterra , Feminino , Seguimentos , Predisposição Genética para Doença/genética , Humanos , Comunicação Interdisciplinar , Colaboração Intersetorial , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Fenótipo , Estudos Prospectivos , Adulto JovemRESUMO
PURPOSE: The present report describes the distribution of breast milk and urinary free and total bisphenol A (BPA) concentrations, from 27 postpartum women and their 31 infants, and explores the influence of age, sex, and nutritional source on infant BPA urinary concentration. METHODS: Both free (unconjugated) and total (free plus conjugated) BPA concentrations from women's breast milk samples and infants' urine samples were measured by online solid-phase extraction coupled to high-performance liquid chromatography-isotope dilution tandem mass spectrometry. Descriptive statistics and nonparametric tests of group comparisons were conducted. RESULTS: Total BPA was detected in 93 % of urine samples in this healthy infant population aged 3-15 months who were without known environmental exposure to BPA [interquartile range (IQR) = 1.2-4.4 µg/L)]. Similarly, 75 % of the mothers' breast milk samples had detectable concentrations of total BPA (IQR = 0.4-1.4 µg/L). The magnitude and frequency of detection of free BPA in the children's urine and the mothers' breast milk were much lower than the total concentrations. CONCLUSIONS: Total BPA was detected in 93 % of this healthy infant population aged 3-15 months who are without known environmental exposure to BPA. Neither free nor total BPA urinary concentrations differed significantly by infant's sex or by nutritional source (breast milk and/or formula) while age group was of borderline significance. There were no significant correlations between free or total BPA concentrations in mothers' breast milk and their infants' urine.
Assuntos
Compostos Benzidrílicos/análise , Monitoramento Ambiental , Poluentes Ambientais/análise , Leite Humano/química , Fenóis/análise , Adulto , Compostos Benzidrílicos/urina , Cromatografia Líquida de Alta Pressão , Poluentes Ambientais/urina , Feminino , Humanos , Lactente , Masculino , Massachusetts , Fenóis/urina , Projetos Piloto , Extração em Fase Sólida , Espectrometria de Massas em TandemRESUMO
Although previous studies suggest nicotine protects against a 6-hydroxydopamine (6-OHDA)-induced lesion of the nigrostriatal tract in rats, it is not known whether functional motor recovery occurs or which nicotinic acetylcholine receptor (nAChR) subtypes mediate this effect. These issues were investigated by comparing the effects of the subtype-specific nAChR agonists, RJR2403 (alpha4beta2 preferring) and (R)-N-(1-azabicyclo[2.2.2.]oct-3-yl)(5-(2-pyridyl)thiopene-2-carboxamide (Compound A; alpha7-selective) and nicotine given 30 min prior to and daily for 14 days after a partial 6-OHDA lesion. In vehicle treated animals, 6-OHDA (6 microg) produced a 65 +/- 1.8% loss of striatal tyrosine hydroxylase (TH) immunoreactivity in the lesion versus intact hemisphere. This loss was reduced in animals treated with nicotine (0.6 and 0.8 mg kg(-1)), reaching significance at the higher dose (36.6 +/- 3.7% loss; P < 0.01 versus vehicle). Treatment with nicotine (0.6 and 0.8 mg kg(-1)) also significantly reduced the number of amphetamine-induced rotations compared to vehicle treatment. In contrast, treatment with RJR2403 (0.2 and 0.4 mg kg(-1)) or Compound A (10 and 20 mg kg(-1)) reduced neither the degree of amphetamine-induced rotations nor the loss of striatal TH immunoreactivity. These data suggest that whilst nicotine is neuroprotective in this partial lesion model, activation of neither the alpha4beta2 nor alpha7 subtypes alone is sufficient to provide protection.
Assuntos
Lesões Encefálicas/prevenção & controle , Feixe Prosencefálico Mediano/patologia , Nicotina/análogos & derivados , Nicotina/uso terapêutico , Agonistas Nicotínicos/uso terapêutico , Oxidopamina , Análise de Variância , Anestésicos Inalatórios/uso terapêutico , Animais , Lesões Encefálicas/induzido quimicamente , Lesões Encefálicas/patologia , Contagem de Células/métodos , Relação Dose-Resposta a Droga , Interações Medicamentosas , Éteres/uso terapêutico , Hidrocarbonetos Fluorados/uso terapêutico , Imuno-Histoquímica/métodos , Masculino , Feixe Prosencefálico Mediano/lesões , Ratos , Ratos Sprague-Dawley , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Whilst local intrastriatal infusion of nicotine consistently elicits striatal dopamine release, systemic administration often fails to do so. Since chronic nicotine administration is known to result in desensitisation-induced upregulation of nicotinic acetylcholine receptors (nAChRs), the present study investigated whether chronic pre-treatment could enhance the response to systemic nicotine and, if so, whether increases in specific nAChR subunit mRNA levels in the substantia nigra pars compacta (SNc) may underlie this effect. In vivo microdialysis studies in male Sprague-Dawley rats revealed that following 4 days pre-treatment with nicotine (0.8 mg kg(-1)s.c.), local intrastriatal nicotine infusion (3 mM) elicited significantly higher dopamine efflux compared to vehicle pre-treated controls (peak release: 1273 +/- 199% basal versus 731 +/- 113% basal), whereas systemic nicotine challenge (0.8 mg kg(-1)s.c.) elicited no response. In contrast, following 8 days pre-treatment with nicotine (0.8 mg kg(-1)s.c.), systemic nicotine challenge (0.8 mg kg(-1)s.c.) now produced significantly higher dopamine efflux than that of vehicle pre-treated controls (147 +/- 30% basal versus 91 +/- 5% basal). Eight days pre-treatment with nicotine also significantly elevated the levels of alpha6 (approximately 55%) and beta3 (approximately 43%) nAChR subunit mRNA in the SNc, suggesting that up-regulation of these nAChR subunit genes in the nigrostriatal tract may contribute to the enhanced nicotine-evoked striatal dopamine release.
Assuntos
Dopamina/metabolismo , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , RNA Mensageiro/biossíntese , Receptores Nicotínicos/metabolismo , Substância Negra/metabolismo , Animais , Autorradiografia , Dopamina/farmacologia , Hibridização In Situ , Masculino , Microdiálise , Oxidopamina , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/biossíntese , Receptores Nicotínicos/efeitos dos fármacos , Substância Negra/efeitos dos fármacos , Simpatectomia Química , SimpatolíticosRESUMO
In the last five years there has been a rapid explosion of publications reporting that neuronal nicotinic acetylcholine receptors (nAChRs) play a role in neurodegenerative disorders. Furthermore, there is a well-established loss of nAChRs in post-mortem brains from patients with Alzheimer's disease, Parkinson's disease and a range of other disorders. In the present review we discuss the evidence that nicotine and subtype selective nAChR ligands can provide neuroprotection in in vitro cell culture systems and in in vivo studies in animal models of such disorders. Whilst in vitro data pertaining to a protective effect of nicotine against nigral neurotoxins like MPTP is less robust, most studies agree that nicotine is protective against glutamate and beta-amyloid toxicity in various culture systems. This effect appears to be mediated by alpha7 subtype nAChRs since the protection is blocked by alpha-bungarotoxin and is mimicked by alpha7 selective agonists. In vivo studies indicate that alpha7 receptors play a critical role in protection from cholinergic lesions and enhancing cognitive function. The exact subtype involved in the neuroprotectant effects seen in animal models of Parkinson's disease is not clear, but in general broad spectrum nAChR agonists appear to provide protection, while alpha4beta2 receptors appear to mediate symptomatic improvements. Evidence favouring a protectant effect of nicotine against acute degenerative conditions is less strong, though some protection has been observed with nicotine pre-treatment in global ischaemia models. A variety of cellular mechanisms ranging from the production of growth factors through to inactivation of toxins and antioxidant actions of nicotine have been proposed to underlie the nAChR-mediated neuroprotection in vitro and in vivo. In summary, although the lack of subtype selective ligands has hampered progress, it is clear that in the future neuronal nAChR agonists could provide functional improvements and slow or halt the progress of several crippling degenerative diseases.
Assuntos
Doenças Neurodegenerativas/patologia , Neurônios/metabolismo , Neurônios/patologia , Receptores Nicotínicos/fisiologia , Doença Aguda , Doença de Alzheimer/patologia , Animais , Encéfalo/patologia , Lesões Encefálicas/patologia , Doença Crônica , Humanos , Doença de Parkinson/patologia , Acidente Vascular Cerebral/patologiaRESUMO
Metabotropic glutamate (mGlu) receptors in the basal ganglia motor loop may increase cell excitability (Group I) or modulate neurotransmitter release (Group I, II and III). Nigrostriatal tract degeneration in Parkinson's disease (PD) produces downstream pathological disturbances in glutamate and GABA transmission. The present study examined whether changes in mGlu receptor gene expression may either contribute to, or compensate for these pathological changes in transmission. In situ hybridisation studies examined the levels of mGlu receptor mRNA in motor loop regions in rats bearing a 6-hydroxydopamine-induced unilateral nigrostriatal tract lesion. Gene expression was reduced in the lesion compared to intact hemispheres for mGlu(1) in the substantia nigra pars compacta (SNc; 51.8+/-11.5%), mGlu(3) in the striatum and globus pallidus (11.7+/-2.8% and 18.9+/-1.4%, respectively) and mGlu(4) in the striatum and premotor cortex (13.8+/-2.7% and 15.8+/-5.5%, respectively). Loss of mGlu(1) mRNA in the SNc confirms that mGlu(1) is highly expressed on dopaminergic neurones where it may contribute to their vulnerability in PD. The down-regulation of mGlu(3) and mGlu(4) mRNA may reflect reduced transcriptional activity in response to increased levels of extracellular glutamate in these regions under parkinsonian conditions. These changes are likely to exacerbate the pathophysiological glutamate and GABA transmission within these regions in PD.
Assuntos
Gânglios da Base/metabolismo , Regulação da Expressão Gênica/fisiologia , Receptores de Glutamato Metabotrópico/biossíntese , Receptores de Glutamato Metabotrópico/genética , Substância Negra/metabolismo , Animais , Corpo Estriado/metabolismo , Masculino , Vias Neurais/metabolismo , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Receptor de Glutamato Metabotrópico 5RESUMO
Loss of striatal dopaminergic innervation in Parkinson's disease (PD) is accompanied by widespread alterations in GABAergic activity within the basal ganglia and thalamus. Accompanying changes in GABA(B) receptor binding have been noted in some basal ganglia regions in parkinsonian primates, suggesting that plasticity of this receptor may also occur in PD. However, the molecular mechanisms underlying the changes in receptor binding and the manner and extent to which different GABA(B) receptor mRNA subunits and splice-variants are affected remain unknown. This study used in situ hybridisation to examine the full profile of changes in expression of the known rat GABA(B) receptor genes and gene variants in the basal ganglia and thalamus of rats, brought about by degeneration of the nigrostriatal tract. All of the GABA(B) mRNA species examined showed unique expression patterns throughout the basal ganglia and thalamus. In addition, all exhibited a marked loss of expression (between 46 and 80%) in the substantia nigra pars compacta of animals bearing a complete 6-hydroxydopamine-induced lesion of the nigrostriatal tract, confirming the presence of these variants in dopaminergic neurones in this region. Further analysis of autoradioagrams revealed additional changes only in GABA(B(1a)) mRNA in discrete anatomical regions. Expression of the GABA(B(1a)) variant was significantly increased in the substantia nigra pars reticulata (33+/-2%), entopeduncular nucleus (26+/-1%) and the subthalamic nucleus (16+/-1%). Since these regions all receive reduced GABAergic innervation following nigrostriatal tract lesioning, it is possible that the increased expression occurs as a compensatory measure. In conclusion, these data demonstrate that GABA(B) receptor genes exhibit regional- and subunit/variant-specific plasticity at the molecular level under parkinsonian conditions.
Assuntos
Gânglios da Base/metabolismo , Expressão Gênica , Receptores de GABA-B/metabolismo , Tálamo/metabolismo , Animais , Autorradiografia , Densitometria , Inibidores da Captação de Dopamina/metabolismo , Masculino , Mazindol/metabolismo , Vias Neurais/efeitos dos fármacos , Sondas de Oligonucleotídeos , Oxidopamina/toxicidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de GABA-B/classificação , Receptores de GABA-B/genética , Substância Negra/efeitos dos fármacos , Trítio/metabolismoRESUMO
In Parkinson's disease, changes in GABAergic activity occurring downstream of the striatal dopamine loss are accompanied by reciprocal changes in GABA(A) receptor binding, the underlying molecular mechanisms for which are unknown. This study examined whether changes in expression of the genes encoding known GABA(A) receptor subunits (alpha(1-4), beta(1-3), gamma(1-3) and delta) could account for this receptor plasticity using a rodent model of Parkinson's disease with a 6-hydroxydopamine-induced nigrostriatal lesion. Analysis of autoradiograms of the basal ganglia and thalamus revealed changes in expression of only four of the 11 subunits studied. Expression of alpha1 and beta2 subunit genes was altered in a parallel manner following a 6-hydroxydopamine lesion; messenger RNA levels for both were significantly increased in the substantia nigra pars reticulata (11 +/- 4% and 17 +/- 1%, respectively), and significantly reduced in the globus pallidus (18 +/- 3% and 16 +/- 3%, respectively) and parafascicular nucleus (19 +/- 3% and 16 +/- 5%, respectively). Smaller changes in the messenger RNA levels encoding the alpha1 subunit in the lateral amygdala (8 +/- 1% decrease) and the alpha4 and gamma2 subunits in the striatum (10 +/- 2% and 6 +/- 1% increase, respectively) were also observed. No changes in expression were noted for any other subunits in any region studied. Clearly, both region- and subunit-specific regulation of GABA(A) receptor subunit gene expression occurs following a nigrostriatal tract lesion. The changes in expression of the alpha1 and beta2 subunit genes probably contribute to the documented changes in GABA(A) receptor binding following striatal dopamine depletion. Moreover, they provide a molecular basis by which the pathological changes in GABAergic activity in Parkinson's disease may be partially compensated.
Assuntos
Gânglios da Base/fisiologia , Corpo Estriado/patologia , Expressão Gênica , Receptores de GABA-A/genética , Substância Negra/patologia , Tálamo/fisiologia , Animais , Autorradiografia , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/fisiopatologia , Masculino , Mazindol/metabolismo , Vias Neurais/efeitos dos fármacos , Vias Neurais/patologia , Oxidopamina/farmacologia , Isoformas de Proteínas/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Substância Negra/efeitos dos fármacos , Substância Negra/fisiopatologia , Distribuição TecidualRESUMO
1 The effect of potassium channel opening compounds has been investigated in the smooth muscle of rabbit aorta under Ca-free conditions. Examination of the characteristics of the response has been performed using cromakalim as the prototype compound. 2 In order of potency, Ro 31-6930, cromakalim, minoxidil sulphate and pinacidil each produced a contraction in rabbit aortic strips bathed in Ca-free MOPS-buffered physiological salt solution (PSS). In contrast, forskolin, glyceryl trinitrate and nifedipine each failed to increase tension under identical conditions. Cromakalim also evoked contraction of bovine trachealis muscle bathed in Ca-free PSS. 3. The contractile response to cromakalim in rabbit aortic strips was of delayed onset (15-20 min) and reached a plateau after approximately 120 min (1.8 g maximum with 1 microM cromakalim). No cromakalim-induced tension changes were observed in either 1 mM or 2.5 mM Ca-containing PSS. 4. Raising the [KCl] of the Ca-free PSS to 65.9 mM fully inhibited the cromakalim-induced contraction in rabbit aortic strips. In addition, pretreatment of aortic strips with the sulphonylurea glibenclamide antagonized the subsequent mechanical response to cromakalim. 5. In Ca-free PSS, cromakalim (1 microM) stimulated 42K-efflux with a time-course corresponding to the contractile event. Glibenclamide (1 microM) inhibited this cromakalim-induced 42K-efflux. 6. In sharp microelectrode studies in bovine trachealis, cromakalim (10 microM) produced a sustained membrane hyperpolarization in normal PSS. In contrast, the cromakalim-induced hyperpolarization in Ca-free PSS was not sustained. The fading of the hyperpolarization was temporally correlated with the increase in tension under these experimental conditions. 7. It is concluded that the K-channel opener-induced smooth muscle contractile response revealed in Ca-free PSS is the consequence of K-channel opening. The nature of the detailed mechanism which underlies this contractile phenomenon remains to be determined.
Assuntos
Benzopiranos/farmacologia , Cálcio/metabolismo , Músculo Liso Vascular/efeitos dos fármacos , Canais de Potássio/efeitos dos fármacos , Pirróis/farmacologia , Vasoconstrição/efeitos dos fármacos , Animais , Aorta/efeitos dos fármacos , Colforsina/farmacologia , Cromakalim , Guanidinas/farmacologia , Masculino , Microeletrodos , Minoxidil/farmacologia , Contração Muscular/efeitos dos fármacos , Nifedipino/farmacologia , Nitroglicerina/farmacologia , Pinacidil , Potássio/metabolismo , Piridinas/farmacologia , CoelhosRESUMO
This study examined whether activation of 5HT(1B) receptors in the rodent globus pallidus (GP) could reduce GABA release in vitro and reverse reserpine-induced akinesia in vivo. Microdissected slices of GP from male Sprague Dawley rats (300-350 g) were preloaded with [(3)H]-GABA. During subsequent superfusion, 4 min fractions were collected for analysis of release. The effects of the 5HT(1B) receptor agonist, 3-(1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3, 2-b]pyrid-5-one (CP-93129), on 25 mM KCl-evoked release were examined using a standard dual stimulation paradigm. Male Sprague Dawley rats (270 - 290 g), stereotaxically cannulated above the GP, were rendered akinetic by injection of reserpine (5 mg kg(-1) s.c.). Eighteen hours later, the rotational behaviour induced by unilateral injection of CP-93129 was examined. CP-93129 (0.6-16.2 microM) produced a concentration-dependent inhibition of 25 mM KCl-evoked [(3)H]-GABA release reaching a maximum inhibition of 52.5+/-4.5%. The effect of a submaximal concentration of CP-93129 (5.4 microM) was fully inhibited by the 5HT(1B) receptor antagonist, isamoltane (10 microM). Following intrapallidal injection, CP-93129 (30-330 nmol in 0.5 microl) produced a dose-dependent increase in net contraversive rotations reaching a maximum of 197+/-32 rotations in 240 min at 330 nmol. Pre-treatment with isamoltane (10 nmol in 1 microl) inhibited the effects of a submaximal dose of CP-93129 (220 nmol) by 84+/-6%. These data suggest that at least some 5HT(1B) receptor function as heteroreceptors in the GP, reducing the release of GABA. Moreover, CP-93129-mediated activation of these receptors in the GP provides relief of akinesia in the reserpine-treated rat model of PD.
Assuntos
Globo Pálido/efeitos dos fármacos , Transtornos dos Movimentos/prevenção & controle , Piridinas/farmacologia , Pirróis/farmacologia , Receptores de Serotonina/efeitos dos fármacos , Agonistas do Receptor de Serotonina/farmacologia , Ácido gama-Aminobutírico/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Globo Pálido/metabolismo , Técnicas In Vitro , Masculino , Transtornos dos Movimentos/etiologia , Propanolaminas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor 5-HT1B de Serotonina , Reserpina/efeitos adversos , Antagonistas da Serotonina/farmacologia , Trítio , Ácido gama-Aminobutírico/metabolismoRESUMO
1. This study examined whether activation of group II metabotropic glutamate (mGlu) receptors in the substantia nigra pars reticulata (SNr) could reverse akinesia in a rodent model of Parkinson's disease (PD). 2. Male Sprague Dawley rats, stereotaxically cannulated above either the SNr or third ventricle, were rendered akinetic by injection of reserpine (5 mg kg-1 s.c.). Eighteen hours later, the rotational behaviour induced by unilateral injection of the group II mGlu receptor agonist, (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV), was examined. 3. Following intranigral injection, DCG-IV (0.125-0.75 nmol in 0.1 microliter) produced a dose-dependent increase in net contraversive rotations (n = 6-8 animals per dose), reaching a maximum of 395 +/- 51 rotations 60 min-1 after 0.75 nmol. The effects of DCG-IV (0.5 nmol) were inhibited by 63.0 +/- 9.0% following 30 min pre-treatment with the group II mGlu receptor antagonist, (2S)-alpha-ethylglutamic acid (EGLU; 100 nmol in 0.2 microliter; n = 6). 4. Following intraventricular injection, DCG-IV (0.125-1.5 nmol in 2 microliters) produced a dose-dependent increase in bilateral locomotor activity (n = 6-7 animals per dose), reaching a maximum of 180 +/- 21 locomotor units 30 min-1 after 0.5 nmol. Pre-treatment with EGLU (200 nmol in 2 microliters) inhibited the effects of DCG-IV (0.5 nmol) by 68.2 +/- 12.3% (n = 5). 5. These data show that activation of group II mGlu receptors in the SNr provides relief of akinesia in the reserpinized rat model of PD. The reversal seen following intraventricular administration supports the likely therapeutic benefit of systemically-active group II mGlu receptor agonists in PD.
Assuntos
Anticonvulsivantes/farmacologia , Antipsicóticos/antagonistas & inibidores , Ciclopropanos/farmacologia , Discinesia Induzida por Medicamentos/tratamento farmacológico , Glicina/análogos & derivados , Receptores de Glutamato Metabotrópico/agonistas , Reserpina/antagonistas & inibidores , Animais , Antipsicóticos/administração & dosagem , Antipsicóticos/toxicidade , Glicina/farmacologia , Injeções , Injeções Intraventriculares , Masculino , Atividade Motora/efeitos dos fármacos , Doença de Parkinson Secundária/induzido quimicamente , Doença de Parkinson Secundária/tratamento farmacológico , Ratos , Ratos Sprague-Dawley , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Reserpina/administração & dosagem , Reserpina/toxicidade , Substância NegraRESUMO
1. Experiments have been performed with the dual intent of analysing the mechanism by which AH 21-132 relaxes airways smooth muscle and determining whether the effects of this compound can be distinguished from those of theophylline. 2. AH 21-132 (0.25-8 microM) and theophylline (1-1000 microM) each caused concentration-dependent suppression of the spontaneous tone of guinea-pig isolated trachealis. The maximal effect of AH 21-132 was equivalent to that of theophylline. No evidence was obtained that the tissue became sensitized or desensitized to the action of AH 21-132. 3. Propranolol (1 microM) profoundly antagonized the tracheal relaxant action of isoprenaline but not that of AH 21-132. 4. In indomethacin (2.8 microM)-treated tissues, tone was induced by K+-rich (120 mM) Krebs solution, acetylcholine (ACh, 1 mM) or histamine (200 microM). Log concentration-relaxation curves for AH 21-132, isoprenaline and theophylline were all moved to the right in the presence of the spasmogens, the smallest rightward shift being induced by histamine and the greatest by ACh. While maximal effects of AH 21-132 and theophylline were unaffected by the spasmogens, that of isoprenaline was reduced by KCl and ACh. 5. In tissues treated with indomethacin (2.8 microM), AH 21-132 (0.1-100 microM) inhibited the spasmogenic effects of potassium chloride (KCl), ACh and histamine in a concentration-dependent manner. The inhibition was characterized by rightward shifts in the spasmogen concentration-effect curves with depression of their maxima. 6. In tissues treated with both indomethacin (2.8 microM) and ACh (1 mM), the removal of tracheal epithelium caused a small but significant leftward shift in the log concentration-relaxation curve for AH 21-132 but did not alter that for theophylline. 7. In tissues treated with indomethacin (2.8 microM) and maintained at 12 degrees C, theophylline (0.1-3.2 mM) caused concentration-dependent spasm. This effect was not shared by AH 21-132. 8. AH 21-132 (0.1-1000 microM) more potently inhibited the activity of cyclic AMP-dependent than of cyclic GMP-dependent phosphodiesterase derived from homogenates of guinea-pig trachealis. Theophylline, too, inhibited these enzymes but was less potent in each case than AH 21-132 and did not exhibit selectivity for the cyclic AMP-dependent enzyme. 9. It is concluded that AH 21-132 exerts a non-specific (i.e. effective no matter what agent is used to support tone) relaxant effect on the trachealis muscle which does not involve the activation of beta l-adrenoceptors. The profile of the relaxant action of AH 21-132 more closely resembles that of theophylline than that of isoprenaline. However, AH 21-132 can be differentiated from theophylline in that: (a) its relaxant potency is increased by epithelial removal; (b) it does not cause tracheal spasm; (c) it exhibits selectivity as an inhibitor of cyclic AMP-dependent as opposed to cyclic GMP-dependent phosphodiesterase. It is possible that the relaxant effects of AH 21-132 are related to its ability to inhibit cyclic nucleotide phosphodiesterases.
Assuntos
Músculo Liso/efeitos dos fármacos , Naftiridinas/farmacologia , Inibidores de Fosfodiesterase/farmacologia , 2',3'-Nucleotídeo Cíclico Fosfodiesterases/antagonistas & inibidores , Acetilcolina/farmacologia , Animais , Feminino , Cobaias , Histamina/farmacologia , Técnicas In Vitro , Isoproterenol/farmacologia , Masculino , Contração Muscular/efeitos dos fármacos , Relaxamento Muscular/efeitos dos fármacos , Tono Muscular/efeitos dos fármacos , Potássio/farmacologia , Teofilina/farmacologia , Traqueia/efeitos dos fármacos , Xantinas/farmacologiaRESUMO
1. The effects of cromakalim on endothelium-denuded rabbit aortic strips were compared with those of the calcium (Ca2+) entry blocking agent, nifedipine. 2. Pre-incubation with cromakalim or nifedipine had no significant effect on the initial phasic component of noradrenaline (NA)-induced responses. 3. Cromakalim (0.3-10 microM), but not nifedipine, inhibited the maintained tonic contractions produced by NA. The effects of cromakalim were antagonized by raising extracellular [K+] or by glibenclamide. 4. Nifedipine inhibited contractions produced by KCl (40 mM) whereas cromakalim had no effect. 5. In Ca2(+)-free physiological salt solution (PSS), cromakalim produced a significant inhibition of both the refilling of and the release of Ca2+ from NA-releasable Ca2+ stores, whereas nifedipine was ineffective. 6. In tissues preloaded with 42K+ cromakalim (0.3-10 microM) produced a concentration-dependent increase in the 42K+ efflux rate coefficient. NA (0.3 microM) also produced an increase in the rate of efflux of 42K+, an effect which was not antagonized by nifedipine (0.3 microM). 7. When microelectrodes were used, cromakalim (1-10 microM) produced a maintained concentration-dependent membrane hyperpolarization. However, low concentrations of cromakalim (less than 1 microM) which relaxed the aorta had no effect on membrane potential. NA had no significant effect on membrane potential. 9. It is concluded that the ability of cromakalim to relax NA-induced contractions in rabbit aorta is not exerted by the indirect closure of nifedipine-sensitive Ca2+ channels. Instead, cromakalim may exert a direct inhibitory action on Ca2+ uptake into and release from Ca2+ stores and additionally inhibit the pathway through which Ca2+ passes from the extracellular fluid to intracellular Ca2+ stores.
Assuntos
Benzopiranos/farmacologia , Nifedipino/farmacologia , Pirróis/farmacologia , Vasodilatação/efeitos dos fármacos , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/fisiologia , Cálcio/metabolismo , Cromakalim , Técnicas In Vitro , Masculino , Potenciais da Membrana/efeitos dos fármacos , Norepinefrina/farmacologia , Potássio/metabolismo , Potássio/farmacologia , Coelhos , Vasodilatação/fisiologiaRESUMO
1. The actions of diazoxide and minoxidil sulphate have been compared with those of cromakalim in rat aorta and portal vein. 2. Diazoxide and minoxidil sulphate hyperpolarized the rat portal vein in a similar manner to cromakalim. 3. Cromakalim, diazoxide and minoxidil sulphate increased 42K and 86Rb efflux from rat portal vein, although minoxidil sulphate had only a small effect on 86Rb efflux. 4. Cromakalim, diazoxide and minoxidil sulphate increased 42K efflux from rat aorta but only cromakalim and diazoxide increased 86Rb efflux from this tissue. 5. Glibenclamide inhibited the relaxant actions of cromakalim, diazoxide and minoxidil sulphate on rat aorta and the increase in 42K efflux produced by these agents in this tissue. 6. Diazoxide relaxed an 80 mM KCl-induced contraction of rat aorta, whilst cromakalim and minoxidil sulphate were without effect. 7. Cromakalim, diazoxide and minoxidil sulphate had no effect on cyclic AMP or cyclic GMP concentrations in rat aorta. 8. It is concluded that diazoxide and minoxidil sulphate like cromakalim exhibit K+ channel opening properties in vascular smooth muscle. Diazoxide exerts an additional inhibitory action not related to the production of cyclic AMP or cyclic GMP. The action of minoxidil sulphate may be primarily located at a K+ channel which is relatively impermeable to 86Rb.
Assuntos
Anti-Hipertensivos/farmacologia , Benzopiranos/farmacologia , Diazóxido/farmacologia , Minoxidil/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Pirróis/farmacologia , Animais , Aorta Torácica/efeitos dos fármacos , Cromakalim , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Glibureto/farmacologia , Técnicas In Vitro , Masculino , Potenciais da Membrana/efeitos dos fármacos , Microeletrodos , Veia Porta/fisiologia , Canais de Potássio/efeitos dos fármacos , Radioisótopos de Potássio , Coelhos , Ratos , Ratos Endogâmicos , Radioisótopos de RubídioRESUMO
(+/-) Pinacidil inhibited noradrenaline-induced contractions in rat aorta and portal vein. The spontaneous tone of guinea-pig bronchial and taenia caeci muscles was relaxed and the spontaneous mechanical activity of rat portal vein was abolished. (+/-) Pinacidil abolished contractions produced by low concentrations of KCl in rat aorta and portal vein, but had relatively little effect on responses to high KCl concentrations. The mechano-inhibitory effects of (+/-) pinacidil were antagonised by tetraethylammonium or procaine. In studies with the purified enantiomers of pinacidil, (-) pinacidil was approximately 20 times more potent that (+) pinacidil. Measurements of electrical activity showed that low concentrations of (+/-) pinacidil selectively inhibited spontaneous electrical discharges in rat portal vein. The duration of multispike electrical complexes was shortened, but spike frequency within a complex and the rate of spike rise and fall were unaffected. At higher concentrations, a dose-dependent hyperpolarisation was observed in both rat aorta and portal vein and the membrane potential approached EK. Using both 86Rb and 42K, (+/-) pinacidil produced a concentration-dependent increase in isotope exchange which correlated with those concentrations at which electrical and mechanical inhibitory effects were observed. Using radioimmunoassay, no pinacidil-induced changes in cyclic AMP or cyclic GMP concentrations were detected in rat aorta. These electrical, ion flux and biochemical measurements suggest that the in vitro mechano-inhibitory effects of pinacidil are associated with the opening of 86Rb-permeable K+ channels in smooth muscle. These effects were observed at concentrations of pinacidil similar to those found in vivo in the plasma of experimental animals and man. It is thus concluded that the hypotensive and antihypertensive effects of pinacidil are the consequence of the cessation of ongoing electrical activity and hyperpolarisation which follows the opening of K+ channels in vascular smooth muscle.
Assuntos
Anti-Hipertensivos/farmacologia , Guanidinas/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Vasodilatadores/farmacologia , Animais , Aorta Torácica/efeitos dos fármacos , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Cobaias , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Norepinefrina/farmacologia , Pinacidil , Veia Porta/efeitos dos fármacos , Potássio/metabolismo , Ratos , Rubídio/metabolismo , Traqueia/efeitos dos fármacosRESUMO
Low-frequency fatigue is a form of muscle fatigue that follows intense muscle activity and is characterized by reduced tetanic tension at low frequencies of stimulation while tetanic tension at high stimulus frequencies is close to normal. The present experiments were performed on isolated single fibers of mouse in which tension and intracellular calcium concentration ([Ca2+]i) were measured. Fatigue was produced by intermittent short tetani continued until tension had declined to 30% of control. Comparison of low- (30- and 50-Hz) and high- (100-Hz) frequency tetani under control conditions and after 30 min of recovery from fatigue showed that low-frequency fatigue was present. During low-frequency fatigue, tetanic [Ca2+]i was substantially reduced at all stimulus frequencies but there was no change in Ca2+ sensitivity or maximum Ca(2+)-activated tension. One possible cause of the reduced tetanic [Ca2+]i is failure of conduction of the action potential in the T tubule, leading to reduced [Ca2+]i in the center of the fiber. However, imaging of [Ca2+]i across the fiber during low-frequency fatigue did not show any such gradient, suggesting that Ca2+ release is uniform across the fiber. Another possible mechanism is that changes in the Ca2+ pumping ability of the sarcoplasmic reticulum might affect tetanic [Ca2+]i. Measurements of the sarcoplasmic reticulum pump function showed a small slowing of Ca2+ uptake rate during low-frequency fatigue, which is unlikely to cause the reduced tetanic [Ca2+]i. In conclusion, the immediate cause of low-frequency fatigue appears to be a reduced tetanic [Ca2+]i, which is probably a consequence of a reduced Ca2+ release from the sarcoplasmic reticulum.
Assuntos
Cálcio/metabolismo , Músculos/metabolismo , Potenciais de Ação/fisiologia , Animais , ATPases Transportadoras de Cálcio/fisiologia , Proteínas Contráteis/fisiologia , Estimulação Elétrica , Fluorescência , Corantes Fluorescentes , Masculino , Camundongos , Contração Muscular/fisiologia , Músculos/citologia , Músculos/fisiologia , Retículo Sarcoplasmático/metabolismoRESUMO
Changes in GABA(A) receptor alpha(1) subunit gene expression occur in the globus pallidus and substantia nigra pars reticulata following lesions of the nigrostriatal tract. To determine whether these changes are translated at the protein level, we performed quantitative autoradiography with the alpha(1) selective ligand, [3H]zolpidem, and the non-selective benzodiazepine site ligand, [3H]Ro 15-1788. Binding of both [3H]zolpidem and [3H]Ro 15-1788 was significantly increased in the substantia nigra pars reticulata (13. 5+/-4.1 and 26.3+/-2.9%, respectively) and significantly reduced in the globus pallidus (20.9+/-0.8 and 18.3+/-1.3%, respectively). These changes in alpha(1) subunit protein expression may help to compensate for the pathological changes in GABAergic activity that occur after striatal dopamine depletion.
Assuntos
Flumazenil/farmacologia , Agonistas GABAérgicos/farmacologia , Moduladores GABAérgicos/farmacologia , Globo Pálido/metabolismo , Piridinas/farmacologia , Substância Negra/metabolismo , Animais , Autorradiografia , Flumazenil/metabolismo , Agonistas GABAérgicos/metabolismo , Moduladores GABAérgicos/metabolismo , Globo Pálido/química , Masculino , Degeneração Neural/metabolismo , Degeneração Neural/patologia , Piridinas/metabolismo , Ensaio Radioligante , Ratos , Ratos Sprague-Dawley , Receptores de GABA-A/fisiologia , Substância Negra/química , Substância Negra/patologia , Trítio , ZolpidemRESUMO
The regional haemodynamic profiles of Ro 31-6930, cromakalim and nifedipine were compared using pulsed Doppler flowmetry in the anaesthetised rat. In order of potency, Ro 31-6930 (0.1-300 micrograms/kg), cromakalim (1-300 micrograms/kg) and nifedipine (1-1000 micrograms/kg) produced dose related falls in mean arterial pressure. The hypotensive effects of Ro 31-6930 and cromakalim were accompanied by reflex tachycardia. All three agents reduced renal vascular resistance by 30-50%. Cromakalim exerted a selective action on this vascular bed. Similar maximal reductions in mesenteric vascular resistance (37-50%) were observed; however, cromakalim was the least potent on this vascular bed. Maximal reductions in iliac vascular resistance (65-78%) were observed, with an order of potency as observed on mean arterial pressure. Qualitative differences in the regional haemodynamic profiles of Ro 31-6930, cromakalim and nifedipine are evident from this study. The different profiles of Ro 31-6930 and cromakalim may reflect structural differences between the pharmacophores of these compounds.
Assuntos
Anti-Hipertensivos/farmacologia , Benzopiranos/farmacologia , Hemodinâmica/efeitos dos fármacos , Nifedipino/farmacologia , Piridinas/farmacologia , Pirróis/farmacologia , Anestesia , Animais , Pressão Sanguínea/efeitos dos fármacos , Cromakalim , Frequência Cardíaca/efeitos dos fármacos , Artéria Ilíaca/efeitos dos fármacos , Masculino , Ratos , Fluxo Sanguíneo Regional/efeitos dos fármacos , Circulação Renal/efeitos dos fármacos , Circulação Esplâncnica/efeitos dos fármacos , Resistência Vascular/efeitos dos fármacosRESUMO
BACKGROUND AND PURPOSE: Increased firing of the glutamatergic pathway between the subthalamic nucleus and substantia nigra pars reticulata (SNpr) contributes to the abnormal firing of motor circuits and subsequent motor deficits seen in Parkinson's disease. Broad spectrum agonist-induced activation of presynaptic group III metabotropic glutamate (mGlu) receptors within the SNpr reduced glutamate release and reversed akinesia in the reserpine-treated rat model of Parkinson's disease. Here, we have sought to identify which subtypes of group III mGlu receptor in the SNpr were responsible for these beneficial effects. EXPERIMENTAL APPROACH: The ability of the mGlu(4) positive allosteric modulator, N-phenyl-7-(hydroxyminocyclopropa[b]chromen-1a-carboxamide) (PHCCC), the mGlu(7) allosteric agonist, N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride (AMN082) and the mGlu(8) -selective agonist (S)-3,4-dicarboxyphenylglycine [(S)-3,4-DCPG] to inhibit KCl-evoked [(3) H]-D-aspartate release was examined in vitro in rat nigral prisms. Reversal of akinesia in reserpine-treated rats was also assessed following intranigral injection of these agents. KEY RESULTS: PHCCC and AMN082 inhibited [(3) H]-D-aspartate release by 42% and 53%, respectively when given alongside a sub-threshold concentration of the broad spectrum group III agonist, L-2-amino-4-phosphonobutyrate (L-AP4; 1 µM). In contrast (S)-3,4-DCPG failed to inhibit [(3) H]-D-aspartate release. All three agents also reversed reserpine-induced akinesia although only the effects of PHCCC and AMN082 were inhibited by pre-treatment with the group III antagonist (RS)-α-cyclopropyl-4-phosphonophenylglycine (CPPG). CONCLUSIONS AND IMPLICATIONS: These findings reveal that targeting SNpr mGlu(4) or mGlu(7) receptors, but not mGlu(8) receptors, provided relief from akinesia in the reserpine-treated rat model of Parkinson's disease, most likely reflecting inhibition of excess glutamate release in this region.
Assuntos
Discinesia Induzida por Medicamentos/metabolismo , Receptores de Glutamato/metabolismo , Substância Negra/metabolismo , Animais , Modelos Animais de Doenças , Discinesia Induzida por Medicamentos/fisiopatologia , Agonistas de Aminoácidos Excitatórios/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Ácido Glutâmico/metabolismo , Masculino , Atividade Motora/efeitos dos fármacos , Doença de Parkinson , Ratos , Ratos Sprague-Dawley , Reserpina , Substância Negra/efeitos dos fármacosRESUMO
BACKGROUND AND PURPOSE: Increased glutamatergic innervation of the substantia nigra pars reticulata (SNpr) and pars compacta (SNpc) may contribute to the motor deficits and neurodegeneration, respectively, in Parkinson's disease (PD). This study aimed to establish whether activation of pre-synaptic group III metabotropic glutamate (mGlu) receptors reduced glutamate release in the SN, and provided symptomatic or neuroprotective relief in animal models of PD. EXPERIMENTAL APPROACH: Broad-spectrum group III mGlu receptor agonists, O-phospho-l-serine (l-SOP) and l-2-amino-4-phosphonobutyrate (l-AP4), were assessed for their ability to inhibit KCl-evoked [(3)H]-d-aspartate release in rat nigral prisms or inhibit KCl-evoked endogenous glutamate release in the SNpr in vivo using microdialysis. Reversal of akinesia in reserpine-treated rats was assessed following intranigral injection of l-SOP and l-AP4. Finally, the neuroprotective effect of 7 days' supra-nigral treatment with l-AP4 was examined in 6-hydroxydopamine (6-OHDA)-lesioned rats. KEY RESULTS: l-SOP and l-AP4 inhibited [(3)H]-d-aspartate release by 33 and 44% respectively. These effects were blocked by the selective group III mGlu antagonist (RS)-alpha-cyclopropyl-4-phosphonophenylglycine (CPPG). l-SOP also reduced glutamate release in the SNpr in vivo by 48%. Injection of l-SOP and l-AP4 into the SNpr reversed reserpine-induced akinesia. Following administration above the SNpc, l-AP4 provided neurochemical, histological and functional protection against 6-OHDA lesion of the nigrostriatal tract. Pretreatment with CPPG inhibited these effects. CONCLUSIONS AND IMPLICATIONS: These findings highlight group III mGlu receptors in the SN as potential targets for providing both symptomatic and neuroprotective relief in PD, and indicate that inhibition of glutamate release in the SN may underlie these effects.