RESUMO
Locus coeruleus degeneration and reduced central noradrenaline content is an early feature of Alzheimer's disease. In transgenic mouse models of Alzheimer's disease-like pathology, lesioning the locus coeruleus exacerbates ß-amyloid (Aß) pathology, neuroinflammation and memory deficits. Here we aimed to determine whether chronic treatment with the α(2)-adrenoceptor antagonist fluparoxan, that enhances noradrenaline release, can prevent the onset of Alzheimer's-like pathology and memory deficits in APP/PS1 transgenic mice (TASTPM). Fluparoxan (1mg/kg/day) was administered to TASTPM and wild type mice from 4 to 8 months of age. Memory was assessed at 4 and 8 months of age using the Morris water maze and contextual fear conditioning and at monthly intervals during the duration of treatment using the object recognition and spontaneous alternation task. Aß plaque load and astrocytosis were measured at 4 and 8 months of age by immunohistochemistry. Fluparoxan treatment prevented age-related spatial working memory deficits in the spontaneous alternation task but not spatial reference memory deficits in the Morris water maze. Aß plaque load and astrocytosis were unaltered by fluparoxan treatment in TASTPM mice. The findings suggest that fluparoxan treatment selectively prevent the decline of forms of memory where noradrenaline plays an integral role and that this beneficial effect is not due to altered Aß plaque pathology or astrocytosis.
Assuntos
Antagonistas de Receptores Adrenérgicos alfa 2/administração & dosagem , Envelhecimento , Peptídeos beta-Amiloides/biossíntese , Gliose/tratamento farmacológico , Transtornos da Memória/prevenção & controle , Piperoxano/análogos & derivados , Placa Amiloide/tratamento farmacológico , Pirróis/administração & dosagem , Envelhecimento/genética , Envelhecimento/metabolismo , Envelhecimento/patologia , Peptídeos beta-Amiloides/genética , Precursor de Proteína beta-Amiloide/biossíntese , Precursor de Proteína beta-Amiloide/genética , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Astrócitos/patologia , Gliose/genética , Gliose/metabolismo , Humanos , Estudos Longitudinais , Transtornos da Memória/genética , Transtornos da Memória/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Piperoxano/administração & dosagem , Placa Amiloide/genética , Placa Amiloide/metabolismo , Presenilina-1/biossíntese , Presenilina-1/genética , Receptores Adrenérgicos alfa 2/fisiologia , Resultado do TratamentoRESUMO
Environmental stress produces adverse affects on memory in humans and rodents. Increased noradrenergic neurotransmission is a major component of the response to stress and noradrenaline (NA) plays an important role in modulating processes involved in learning and memory. The present study investigated the effect of NA depletion on stress-induced changes on memory performance in the mouse. Central NA depletion was induced using the selective neurotoxin N-(2-chloroethyl)-N-ethyl-2 bromobenzylamine (DSP-4) and verified by high performance liquid chromatography (HPLC). A novel cage stress procedure involving exposure to a new clean cage for 1 h per day, 4 days per week for 4 weeks, was used to produce stress-induced memory deficits measured using the object recognition task. 50 mg/kg DSP-4 produced large and sustained reductions in NA levels in the frontal cortex and hippocampus measured 24 h, 1 week and 5 weeks after treatment. Four weeks of exposure to novel cage stress induced a memory deficit in the object recognition task which was prevented by DSP-4 pre-treatment (50 mg/kg 1 week before the commencement of stress).These findings indicate that chronic environmental stress adversely affects recognition memory and that this effect is, in part, mediated by the noradrenergic stress response. The implication of these findings is that drugs targeting the noradrenergic system to reduce over-activity may be beneficial in the treatment of stress-related mental disorders such as post-traumatic stress disorder or anxiety in which memory is affected.
Assuntos
Adrenérgicos/farmacologia , Benzilaminas/farmacologia , Transtornos da Memória/tratamento farmacológico , Transtornos da Memória/etiologia , Norepinefrina/metabolismo , Estresse Psicológico/complicações , Animais , Meio Ambiente , Lobo Frontal/efeitos dos fármacos , Lobo Frontal/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Ácido Hidroxi-Indolacético/metabolismo , Masculino , Transtornos da Memória/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Testes Neuropsicológicos , Distribuição Aleatória , Reconhecimento Psicológico/efeitos dos fármacos , Reconhecimento Psicológico/fisiologia , Serotonina/metabolismo , Estresse Psicológico/metabolismo , Fatores de TempoRESUMO
Orexin A, synthesised in the posterolateral hypothalamus, has widespread distribution including the paraventricular nucleus (PVN), which is rich in thyrotropin-releasing hormone (TRH) neurones. Nerve fibres in the PVN synapse on neurones that send polysynaptic projections to brown adipose tissue (BAT), which is important in thermogenesis. A number of observations suggests orexin A may be involved in regulation of metabolism and thermogenesis. We investigated the effect of orexin A injected intracerebroventricularly (ICV) on thyroid-stimulating hormone (TSH) and thyroid hormones in male rats. We then examined the effect of chronic iPVN injections of orexin A on plasma TSH and uncoupling protein-1 (UCP-1) protein in BAT. Orexin A (3 nmol) administered ICV significantly suppressed plasma TSH at 10 and 90 min. Orexin A (0.3 nmol) administered into the PVN twice daily for 3 days significantly increased day-time 2-h food intake, but did not significantly alter nocturnal food intake. Though chronic iPVN orexin A altered diurnal food intake, there was no effect on 24-h food intake or body weight. Furthermore, orexin A administered chronically into the PVN did not alter UCP-1 level in BAT, or plasma hormones relative to saline injected animals. Chronic iPVN orexin A does not appear to influence thermogenesis through activation of UCP-1 or the thyroid axis.
Assuntos
Tecido Adiposo Marrom/efeitos dos fármacos , Proteínas de Transporte/metabolismo , Proteínas de Transporte/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas de Membrana/metabolismo , Neuropeptídeos/farmacologia , Glândula Tireoide/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Proteínas de Transporte/administração & dosagem , Injeções Intraventriculares/métodos , Canais Iônicos , Masculino , Proteínas Mitocondriais , Neuropeptídeos/administração & dosagem , Orexinas , Hormônios Hipofisários/sangue , Ratos , Ratos Wistar , Tireotropina/sangue , Tiroxina/sangue , Tri-Iodotironina/sangue , Proteína Desacopladora 1RESUMO
Neuropeptide FF (NPFF) is a mammalian peptide that is found in high concentrations in the central nervous system (CNS) and has also been detected in plasma. Various functions have been attributed to this peptide although its main action in the CNS remains unclear. In this study we observed that intracerebroventricular (ICV) injection of human NPFF, at early light phase in fasted rats, acutely reduced food intake and caused a large increase in water intake compared with saline injected controls. This effect was independently observed in two separate studies yielding similar results. Thus the central effects of NPFF to decrease food intake may be largely attributable to increased water intake.
Assuntos
Ingestão de Líquidos/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Proteínas Imediatamente Precoces , Antagonistas de Entorpecentes/administração & dosagem , Oligopeptídeos/administração & dosagem , Animais , Comportamento Animal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Proteínas de Ligação a DNA/metabolismo , Relação Dose-Resposta a Droga , Proteína 1 de Resposta de Crescimento Precoce , Humanos , Injeções Intraventriculares , Masculino , Atividade Motora/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Tempo de Reação , Fatores de Tempo , Fatores de Transcrição/metabolismoRESUMO
The orexins are neuropeptides originally reported to be involved in the stimulation of food intake. However, analysis of orexin immunoreactive fibres have revealed the densest innervation in brain sites involved in arousal and sleep-wake control, notably the noradrenergic locus coeruleus, an area that also expresses orexin receptor 1 (OX1R) messenger RNA (mRNA). We report here that, in the rat, a single intracerebroventricular injection of orexin A (1 and 3 nmol) or orexin B (3 nmol), during the early light phase, did not increase food intake over the first 4 h postinjection. However, the frequency of active behaviors such as grooming, rearing, burrowing and locomotion increased. Feeding behavior and food intake subsequently decreased over the following 20 h (4-24 h postinjection period) in the orexin A 3 nmol injected group whilst the frequency of inactive behavior (still or asleep) in this group increased. Using riboprobes, we performed in situ hybridization histochemistry to map the distribution of orexin receptor 2 (OX2R) mRNA within the rat brainstem. We report here, for the first time, the presence of OX2R mRNA in the nucleus of the solitary tract and the lateral reticular field (LRt). The LRt is a brainstem site that, amongst other functions, is implicated in attention and wakefulness. This distribution of OX2R and the effects on behavior support recent reports that the orexins might modulate central nervous system arousal and sleep-wake mechanisms rather than exclusively being involved in the control of food intake.
Assuntos
Apetite/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Tronco Encefálico/efeitos dos fármacos , Proteínas de Transporte/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas do Tecido Nervoso/genética , Neuropeptídeos/farmacologia , RNA Mensageiro/análise , Receptores de Neuropeptídeos/genética , Animais , Apetite/fisiologia , Atenção/efeitos dos fármacos , Atenção/fisiologia , Mapeamento Encefálico , Tronco Encefálico/química , Tronco Encefálico/fisiologia , Proteínas de Transporte/administração & dosagem , Ritmo Circadiano/efeitos dos fármacos , Ingestão de Alimentos/fisiologia , Regulação da Expressão Gênica/efeitos da radiação , Asseio Animal/efeitos dos fármacos , Hipotálamo/fisiologia , Hibridização In Situ , Injeções Intraventriculares , Luz , Masculino , Atividade Motora/efeitos dos fármacos , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/efeitos dos fármacos , Proteínas do Tecido Nervoso/fisiologia , Neuropeptídeos/administração & dosagem , Receptores de Orexina , Orexinas , Especificidade de Órgãos , Ratos , Ratos Wistar , Receptores Acoplados a Proteínas G , Receptores de Neuropeptídeos/biossíntese , Receptores de Neuropeptídeos/efeitos dos fármacos , Receptores de Neuropeptídeos/fisiologia , Sono/efeitos dos fármacos , Vigília/efeitos dos fármacos , Vigília/fisiologiaRESUMO
Cocaine- and amphetamine-regulated transcript (CART) and CART peptide are abundant in hypothalamic nuclei controlling anterior pituitary function. Intracerebroventricular (ICV) injection of CART peptide results in neuronal activation in the paraventricular nucleus (PVN), rich in corticotrophin-releasing factor (CRH) and thyrotrophin-releasing factor (TRH) immunoreactive neurons. The aims of this study were three-fold. Firstly, to examine the effects of CART peptide on hypothalamic releasing factors in vitro, secondly, to examine the effect of ICV injection of CART peptide on plasma pituitary hormones and finally to examine the effect of PVN injection of CART peptide on food intake and circulating pituitary hormones. CART(55-102) (100 nM) peptide significantly stimulated the release of CRH, TRH and neuropeptide Y from hypothalamic explants but significantly reduced alpha melanocyte stimulating hormone release in vitro. Following ICV injection of 0.2 nmol CART(55-102), a dose which significantly reduces food intake, plasma prolactin (PRL), growth hormone (GH) and adrenocorticotrophin hormone (ACTH) and corticosterone increased significantly. Following PVN injection of CART(55-102), food intake was significantly reduced only at 0.2 and 0.6 nmol. However, PVN injection of 0.02 nmol CART(55-102) produced a significant increase in plasma ACTH. ICV injection of CART peptide significantly reduces food intake. Unlike many anorexigenic peptides, there is no increased sensitivity to PVN injection of CART(55-102). In contrast, both ICV and PVN injection of CART(55-102) significantly increased plasma ACTH and release of hypothalamic CRH is significantly increased by CART peptide in vitro. This suggests that CART peptide may play a role in the control of pituitary function and in particular the hypothalamo-pituitary adrenal axis.
Assuntos
Regulação do Apetite/efeitos dos fármacos , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Proteínas do Tecido Nervoso/administração & dosagem , Animais , Hormônio Liberador da Corticotropina/metabolismo , Glucose/administração & dosagem , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Técnicas In Vitro , Injeções Intraventriculares , Masculino , Microinjeções , Proteínas do Tecido Nervoso/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neuropeptídeo Y/metabolismo , Neuropeptídeos/metabolismo , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Fragmentos de Peptídeos/administração & dosagem , Hormônios Hipofisários/sangue , Ratos , Ratos Wistar , Proteínas Recombinantes/administração & dosagem , Organismos Livres de Patógenos Específicos , Hormônio Liberador de Tireotropina/metabolismoRESUMO
The melanocortin-4 receptor (MC4R) in the hypothalamus is thought to be important in physiological regulation of food intake. We investigated which hypothalamic areas known to express MC4R are involved in the regulation of feeding by using alpha-melanocyte-stimulating hormone (alpha-MSH), an endogenous MC4R agonist, and agouti-related peptide (Agrp), an endogenous MC4R antagonist. Cannulae were inserted into the rat hypothalamic paraventricular (PVN), arcuate (Arc), dorsomedial (DMN), and ventromedial (VMN) nuclei; the medial preoptic (MPO), anterior hypothalamic (AHA), and lateral hypothalamic (LHA) areas; and the extrahypothalamic central nucleus of the amygdala (CeA). Agrp (83-132) (0.1 nmol) and [Nle4, D-Phe7]alpha(-MSH (NDP-MSH) (0.1 nmol), a stable alpha-MSH analog, were administered to fed and fasted rats, respectively. The PVN, DMN, and MPO were the areas with the greatest response to Agrp and NDP-MSH. At 8 h postinjection, Agrp increased feeding in the PVN by 218 +/- 23% (P < 0.005), in the DMN by 268 +/- 42% (P < 0.005), and in the MPO by 236 +/- 31% (P < 0.01) compared with a saline control group for each nucleus. NDP-MSH decreased food intake in the PVN by 52 +/- 6% (P < 0.005), in the DMN by 44 +/- 6% (P < 0.0001), and in the MPO by 55 +/- 6% (P < 0.0001) at 1 h postinjection. Injection into the AHA and CeA resulted in smaller alterations in food intake. No changes in feeding were seen after the administration of Agrp into the Arc, LHA, or VMN, but NDP-MSH suppressed food intake in the Arc and LHA. This study indicates that the hypothalamic nuclei expressing MC4R vary in their sensitivity to Agrp and alpha-MSH with regard to their effect on feeding.
Assuntos
Ingestão de Alimentos/fisiologia , Hipotálamo/fisiologia , Proteínas/fisiologia , alfa-MSH/fisiologia , Proteína Relacionada com Agouti , Animais , Mapeamento Encefálico , Ingestão de Alimentos/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Injeções Intraventriculares , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Fragmentos de Peptídeos/farmacologia , Proteínas/química , Proteínas/farmacologia , Ratos , Ratos Wistar , alfa-MSH/análogos & derivados , alfa-MSH/farmacologiaRESUMO
The melanocortin-4 receptor (MC4-R) appears to be an important downstream mediator of the action of leptin. We examined to what extent the anorectic effects of cocaine- and amphetamine-regulated transcript (CART), glucagon-like peptide-1 (GLP-1) and corticotrophin releasing factor (CRF) might be mediated via MC4-R. alpha-Melanocyte stimulating hormone (alpha-MSH), the MC4-R agonist, administered intracerebroventricularly (ICV) at a dose of 1 nmol reduced food intake by approximately half. Agouti-related protein (Agrp) (83-132), a biologically active fragment of the endogenous MC4-R antagonist, administered ICV at a dose of 1 nmol completely blocked the anorectic effect of 1 nmol alpha-MSH. CART (55-102) (0.2 nmol), GLP-1 (3 nmol) and CRF (0.3 nmol) produced a reduction in feeding of approximately the same magnitude as 1 nmol alpha-MSH. Agrp (83-132) (1 nmol) administered ICV did not block the anorectic effects of CART (55-102) (1 h food intake, 0.2 nmol CART (55-102), 2.7+/-0.8 g vs. CART (55-102)+Agrp (83-132), 2.6+/-0.6 g, P=0.87; saline control 5.4+/-0.3 g, P<0.001 vs. both groups). Agrp (83-132) also did not block the anorectic effects of GLP-1 or CRF (1 h food intake, 0.3 nmol CRF, 0.7+/-0.3 g vs. CRF+Agrp (83-132), 0.7+/-0.3 g, P=0.91; 3 nmol GLP-1, 1.9+/-0.4 g vs. GLP-1+Agrp (83-132), 1.1+/-0. 5 g, P=0.23; saline control 5.0+/-0.6 g, P<0.001 vs. all four groups). Thus, as previous data suggests, GLP-1 and CRF do not appear to reduce food intake predominantly via MC4-R, we here demonstrate for the first time that CART, in addition to GLP-1 and CRF primarily acts via Agrp independent pathways.
Assuntos
Hormônio Liberador da Corticotropina/metabolismo , Ingestão de Alimentos/efeitos dos fármacos , Glucagon/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Fragmentos de Peptídeos/metabolismo , Precursores de Proteínas/metabolismo , Proteínas/metabolismo , Receptores de Peptídeos/efeitos dos fármacos , Proteína Relacionada com Agouti , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Hormônio Liberador da Corticotropina/farmacologia , Ingestão de Alimentos/fisiologia , Glucagon/farmacologia , Peptídeo 1 Semelhante ao Glucagon , Peptídeos e Proteínas de Sinalização Intercelular , Leptina/metabolismo , Masculino , Proteínas do Tecido Nervoso/farmacologia , Fragmentos de Peptídeos/farmacologia , Precursores de Proteínas/farmacologia , Proteínas/farmacologia , Ratos , Ratos Wistar , Receptor Tipo 4 de Melanocortina , Receptores de Peptídeos/antagonistas & inibidores , Receptores de Peptídeos/metabolismo , alfa-MSH/farmacologiaRESUMO
Prolonged fasting is associated with a downregulation of the hypothalamo-pituitary thyroid (H-P-T) axis, which is reversed by administration of leptin. The hypothalamic melanocortin system regulates energy balance and mediates a number of central effects of leptin. In this study, we show that hypothalamic melanocortins can stimulate the thyroid axis and that their antagonist, agouti-related peptide (Agrp), can inhibit it. Intracerebroventricular (ICV) administration of Agrp (83-132) decreased plasma thyroid stimulating hormone (TSH) in fed male rats. Intraparaventricular nuclear administration of Agrp (83-132) produced a long-lasting suppression of plasma TSH, and plasma T4. ICV administration of a stable alpha-MSH analogue increased plasma TSH in 24-hour-fasted rats. In vitro, alpha-MSH increased thyrotropin releasing hormone (TRH) release from hypothalamic explants. Agrp (83-132) alone caused no change in TRH release but antagonized the effect of alpha-MSH on TRH release. Leptin increased TRH release from hypothalami harvested from 48-hour-fasted rats. Agrp (83-132) blocked this effect. These data suggest a role for the hypothalamic melanocortin system in the fasting-induced suppression of the H-P-T axis.
Assuntos
Hipotálamo/metabolismo , Leptina/metabolismo , Hipófise/metabolismo , Receptores da Corticotropina/metabolismo , Glândula Tireoide/metabolismo , Tireotropina/sangue , Proteína Relacionada com Agouti , Animais , Jejum , Hipotálamo/efeitos dos fármacos , Injeções , Peptídeos e Proteínas de Sinalização Intercelular , Leptina/farmacologia , Masculino , Proteínas/administração & dosagem , Proteínas/metabolismo , Ratos , Ratos Wistar , Receptores da Corticotropina/antagonistas & inibidores , Receptores de Melanocortina , alfa-MSH/administração & dosagem , alfa-MSH/análogos & derivados , alfa-MSH/metabolismoRESUMO
Orexins are a family of neuropeptides originally believed to be important mediators of food intake. The wide distribution of orexins and their receptors, however, has suggested other regulatory functions for these peptides including involvement in sleep and arousal mechanisms. In this study, we have demonstrated diurnal variation in orexin A immunoreactivity in the pons, from where locus coeruleus noradrenergic neurones innervate other brain areas to stimulate arousal, and in the preoptic/anterior hypothalamic region, an area implicated in the regulation of sleep and circadian rhythms. Orexin A immunoreactivity decreased by 50% in the preoptic/anterior hypothalamus from 09:00 to 21:00 h (P < 0.0001), whilst in the pons, it increased by over 30% from 09:00 to 01:00 h (P = 0.02). Prepro-orexin mRNA also displayed diurnal variation. This further suggests that orexins are involved in the regulation of the sleep/wake cycle.
Assuntos
Encéfalo/metabolismo , Proteínas de Transporte/metabolismo , Ritmo Circadiano/fisiologia , Regulação da Expressão Gênica/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular , Neuropeptídeos/metabolismo , Precursores de Proteínas/genética , Transcrição Gênica , Animais , Nível de Alerta , Proteínas de Transporte/análise , Hipotálamo Anterior/metabolismo , Locus Cerúleo/metabolismo , Masculino , Neuropeptídeos/análise , Orexinas , Ponte/metabolismo , Área Pré-Óptica/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/genética , Ratos , Ratos Wistar , SonoRESUMO
Orexin-A and orexin-B (the hypocretins) are recently described neuropeptides suggested to have a physiological role in the regulation of food intake in the rat. We compared the orexigenic effect of the orexins administered intracerebroventricular (ICV) with other known stimulants of food intake, one strong, neuropeptide Y (NPY), and two weaker, melanin-concentrating hormone (MCH) and galanin. Orexin-A consistently stimulated food intake, but orexin-B only on occasions. Both peptides stimulated food intake significantly less than NPY, but to a similar extent to MCH (2 h food intake: NPY 3 nmol, 7.2+/-0.9 g vs saline, 1.5+/-0.2 g, P<0.001, MCH 3 nmol, 3.2+/-0.8 g vs saline, P<0.01, orexin-B 30 nmol, 2. 6+/-0.5 g vs saline, P=0.11) and to galanin (1 h food intake: galanin 3 nmol, 2.0+/-0.4 g vs saline, 0.8+/-0.2 g, P<0.05, orexin-A 3 nmol 2.2+/-0.4 g vs saline, P<0.01; 2 hour food intake: orexin-B 3 nmol, 2.4+/-0.3 g vs saline, 1.3+/-0.2 g, P<0.05). Following ICV orexin-A, hypothalamic c-fos, a maker of neuronal activation, was highly expressed in the paraventricular nucleus (PVN), and the arcuate nucleus (P<0.005 for both). IntraPVN injection of orexin-A stimulated 2 h food intake by one gram (orexin-A 0.03 nmol, 1.6+/-0. 3 g vs saline, 0.5+/-0.3 g, P<0.005). These findings support the suggestion that the orexins stimulate food intake. However, this effect is weak and may cast doubt upon their physiological importance in appetite regulation in the rat.
Assuntos
Ingestão de Alimentos/efeitos dos fármacos , Hipotálamo/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Neuropeptídeos/farmacologia , Animais , Proteínas de Transporte/farmacologia , Galanina/farmacologia , Expressão Gênica/efeitos dos fármacos , Genes fos , Hormônios Hipotalâmicos/farmacologia , Masculino , Melaninas/farmacologia , Neuropeptídeo Y/farmacologia , Orexinas , Hormônios Hipofisários/farmacologia , Ratos , Ratos WistarRESUMO
Central nervous system glucagon-like peptide-1-(7-36) amide (GLP-1) administration has been reported to acutely reduce food intake in the rat. We here report that repeated intracerebroventricular (i.c.v.) injection of GLP-1 or the GLP-1 receptor antagonist, exendin-(9-39), affects food intake and body weight. Daily i.c.v. injection of 3 nmol GLP-1 to schedule-fed rats for 6 days caused a reduction in food intake and a decrease in body weight of 16 +/- 5 g (P < 0.02 compared with saline-injected controls). Daily i.c.v. administration of 30 nmol exendin-(9-39) to schedule-fed rats for 3 days caused an increase in food intake and increased body weight by 7 +/- 2 g (P < 0.02 compared with saline-injected controls). Twice daily i.c.v. injections of 30 nmol exendin-(9-39) with 2.4 nmol neuropeptide Y to ad libitum-fed rats for 8 days increased food intake and increased body weight by 28 +/- 4 g compared with 14 +/- 3 g in neuropeptide Y-injected controls (P < 0.02). There was no evidence of tachyphylaxis in response to i.c.v. GLP-1 or exendin-(9-39). GLP-1 may thus be involved in the regulation of body weight in the rat.
Assuntos
Peso Corporal/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Animais , Ingestão de Energia/efeitos dos fármacos , Glucagon , Peptídeo 1 Semelhante ao Glucagon , Peptídeos Semelhantes ao Glucagon , Injeções Intraventriculares , Masculino , Neurotransmissores/administração & dosagem , Neurotransmissores/farmacologia , Fragmentos de Peptídeos/administração & dosagem , Ratos , Ratos WistarRESUMO
Agouti-related protein (Agrp) is present in rat and human hypothalamus and is structurally related to agouti protein. Overexpression of either of these proteins results in obesity. However the effect of exogenous Agrp and its in vivo interaction with alpha-melanocyte stimulating hormone (alphaMSH), the likely endogenous melanocortin 3 and 4 receptor (MC3-R and MC4-R) agonist, have not been demonstrated. We report that 1 nmol of Agrp(83-132), a C-terminal fragment of Agrp, when administered intracerebroventricularly (ICV) into rats, increased food intake over a 24-h period (23.0+/-1.4 g saline vs 32.9+/-2.3 g Agrp, p<0.05). The hyperphagia was similar to that seen when 1 nmol of the synthetic MC3-R and MC4-R antagonist SHU9119 was given i.c.v. (19.6+/-1.8 g saline vs 32.5+/-1.7 g SHU9119, p<0.001). Both Agrp(83-132) and SHU9119 blocked the reduction in 1-h food intake of i.c.v. alphaMSH at the beginning of the dark phase. This effect occurred independently of whether the antagonists were administered simultaneously, or nine hours prior, to the alphaMSH. We have also shown Agrp(83-132) is an antagonist at the MC3-R and MC4-R, with similar inhibition of cAMP activation to that previously reported for the full length peptide. In conclusion, Agrp(83-132) administered i.c.v. increases feeding with long lasting effects and is able to inhibit the action of alphaMSH. This interaction may be mediated by the MC3-R and/or MC4-R.
Assuntos
Ingestão de Alimentos/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular , Fragmentos de Peptídeos/farmacologia , Proteínas/farmacologia , alfa-MSH/antagonistas & inibidores , Proteína Agouti Sinalizadora , Proteína Relacionada com Agouti , Animais , Relação Dose-Resposta a Droga , Injeções Intraventriculares , Masculino , Ratos , Ratos Wistar , Receptor Tipo 3 de Melanocortina , Receptores da Corticotropina/efeitos dos fármacosRESUMO
The title compound, (+)-MCPG [(+)-alpha-(4-carboxyphenyl)-alpha-methylglycine, C10H11NO4], is an antagonist at certain subtypes of metabotropic glutamate (mGlu) receptors. (+)-MCPG has gained widespread acceptance as a tool for probing the physiological role of mGlu receptors in the central nervous system. As a result, mGlu receptors are now known to be involved in processes connected with learning and memory, modulation of synaptic transmission and the transmission of pain responses. (+)-MCPG crystallized in its zwitterionic form. Its absolute configuration was assigned as S from X-ray diffraction data collected at 150 K. The refined Flack parameter is consistent with this assignment, although the large e.s.d. associated with it introduces some ambiguity.
Assuntos
Benzoatos/química , Antagonistas de Aminoácidos Excitatórios/química , Glicina/análogos & derivados , Cristalização , Glicina/química , Difração de Raios XRESUMO
All four isomers of 3-(4-chlorophenyl)glutamic acid (5-8) were prepared by diastereoselective synthesis. Addition of (6S)-(+)-bis-lactim ether 15 to cis-4-chlorocinnamate 12 gave a mixture comprising mainly the (2R,3S)- and (2R,3R)-isomers 5 and 6, respectively (in a ratio of 56:40), while addition of (6R)-(-)-bis-lactim ether 16 to 4-chlorocinnamate 12 gave a mixture comprising mainly the (2S,3R)- and (2S,3S)-isomers 8 and 7, respectively (in a ratio of 56:42). The four stereoisomers (5-8) were therefore conveniently prepared by addition of either 3-lithio-(6S)- or -(6R)-bis-lactim ether (15 or 16, respectively) to 4-chlorocinnamate 12 and separation of the resultant mixtures of diastereoisomers (23-26) by flash silica gel chromatography. The absolute configurations of 6 and 7 were confirmed by X-ray crystallography. Both the (2S,3S)- and (2S,3R)-isomers (7 and 8, respectively) at a concentration of 100 microM significantly potentiated depolarizations induced by 10 microM L-homocysteic acid (L-HCA) (% control +/- sem: 130.4 +/- 3.6, n = 20 and 114.5 +/- 2.4, n = 11, respectively) while the (2R,3S)-isomer 5 significantly reduced L-HCA response amplitude (94.2 +/- 1.4, n = 9) and the (2R,3R)-isomer 6 was inactive. Experiments designed to compare the agonist-potentiating actions of 7 and 8 in the neonatal rat spinal cord with L-trans-pyrrolidine-2,4-dicarboxylic acid, the well-known L-Glu uptake inhibitor, provided additional evidence for the selective enhancement of depolarizations due to L-HCA and not those due to L-Glu. This selective action supports the existence of multiple excitatory amino acid uptake sites.
Assuntos
Glutamatos/síntese química , Glutamatos/farmacologia , Ácido Glutâmico/análogos & derivados , Homocisteína/análogos & derivados , Animais , Cristalografia por Raios X , Eletrofisiologia , Ácido Glutâmico/metabolismo , Homocisteína/farmacologia , Ligação de Hidrogênio , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Neurônios/efeitos dos fármacos , Neurotransmissores/química , Neurotransmissores/metabolismo , Ratos , Medula Espinal , EstereoisomerismoRESUMO
The four recently synthesized stereoisomers of 3-(4-chlorophenyl) glutamic acid (chlorpheg) were individually examined for their abilities to potentiate depolarizations of neonatal rat motoneurones evoked by L-homocysteic acid (L-HCA, 10 microM). This property had previously been observed using the racemate and is believed to be mediated by uptake inhibition. Both the (2S,3S)- and (2S,3R)- isomers were selective potentiators of L-HCA- (vs L-Glu) induced depolarizations although the (2S,3S)- isomer was more effective. The (2R,3S)- isomer had a slight but significant depressant action which could be attributed to N-methyl-D-aspartate (NMDA) receptor antagonism. Comparison of the potentiating properties of (2S,3S)- and (2S,3R)-chlorpheg with those of L-trans-pyrrolidine-2,4-dicarboxylic acid (tPDC, a L-Glu uptake inhibitor) upon L-HCA- and L-Glu-evoked responses revealed that both chlorpheg isomers (500 microM each) selectively potentiated responses evoked by L-HCA (10 microM) but had no significant effect upon those evoked by L-Glu (50 microM). On the other hand, use of tPDC at the same concentration significantly enhanced the depolarizations evoked by both amino acids, although its action on L-Glu-evoked responses was greater. It is concluded that (i) the (2S,3S)- isomer and to a lesser extent, the (2S,3R)- isomer of chlorpheg are responsible for the potentiating actions seen with the chlorpheg racemate used in previous studies and (ii) (2R,3S)-chlorpheg is a weak NMDA antagonist. The apparently selective action of (2S,3S)- and (2S,3R)-chlorpheg upon L-HCA-relative to L-Glu-induced depolarizations supports the existence of multiple excitatory amino acid uptake sites, some of which may yet be unidentified.
Assuntos
Ácidos Dicarboxílicos/farmacologia , Glutamatos/farmacologia , Neurônios Motores/efeitos dos fármacos , Pirrolidinas/farmacologia , Medula Espinal/efeitos dos fármacos , Animais , Interações Medicamentosas , Ácido Glutâmico , Homocisteína/análogos & derivados , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Ratos , EstereoisomerismoRESUMO
1. This study examined the binding of the new radioligand (S)-[3H]-5-fluorowillardiine to rat brain synaptic membranes. Specific binding represented greater than 80% of the total binding and was increased by 10% in the presence of 100mM potassium thiocyanate (KSCN). 2. In the absence of KSCN, (S)-[3H]-5-fluorowillardiine identified two binding sites with KD1=22.5 nM, Bmax1=1.4 pmol mg(-1) protein and KD2=1.5 microM, Bmax2=10.8 pmol mg(-1) protein. In the presence of 100 mM KSCN the affinities of both the binding sites were increased, yielding values of KD1=6.9 nM and KD2=0.4 microM KSCN was without effect on the Bmax values. 3. (S)-[3H]-5-fluorowillardiine binding was displaced by non-NMDA receptor ligands with the rank order of potency: 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo(F)quinoxaline (NBQX) > domoate > (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionice acid (AMPA) = L-glutamate > 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) > kainate >> (R)-5-fluorowellardiine. In contrast, both N-methyl-D-aspartate (NMDA) and the metabotropic glutamate receptor agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) were inactive. 4. By use of quantitative autoradiography the regional distribution of (S)-[3H]-5-fluorowillardiine binding in rat brain was assessed. The highest levels of binding were in the dentate gyrus and the CA1 region of the hippocampus. Lower levels of binding were detected in the cerebral cortex, olfactory system, lateral septum, caudate putamen and nucleus accumbens. 5. We conclude that the pharmacological profile and regional distribution of (S)-[3H]-5-fluorowillardiine binding is consistent with its specific interaction with AMPA receptors.
Assuntos
Alanina/análogos & derivados , Encéfalo/efeitos dos fármacos , Agonistas de Aminoácidos Excitatórios/farmacologia , Pirimidinas/farmacologia , Receptores de AMPA/efeitos dos fármacos , Alanina/metabolismo , Alanina/farmacologia , Animais , Autorradiografia , Encéfalo/metabolismo , Agonistas de Aminoácidos Excitatórios/metabolismo , Técnicas In Vitro , Masculino , Pirimidinas/metabolismo , Ensaio Radioligante , Ratos , Ratos Wistar , Membranas Sinápticas/efeitos dos fármacos , Membranas Sinápticas/metabolismo , Distribuição TecidualRESUMO
The depression of the monosynaptic excitation of neonatal rat motoneurones produced by the metabotropic glutamate receptor (mGluR) agonists (1S,3S)-1-aminocyclopentane-1, 3-dicarboxylate (ACPD) or L-2-amino-4-phosphonobutyrate (L-AP4) was antagonized by three novel phenylglycine analogues: (RS)-alpha-methyl-4-sulphonophenylglycine (MSPG), (RS)-alpha-methyl-4-phosphonophenylglycine (MPPG) and (RS)-alpha-methyl-4-tetrazolylphenylglycine (MTPG). The potencies of all the new compounds were greater than that of the previously reported (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG). For L-AP4-sensitive presynaptic mGluRs, the order of antagonist potency found was MPPG > MSPG > MTPG > MCPG. In contrast, the order of antagonist potency found for (1S,3S)-ACPD-sensitive presynaptic mGluRs was MTPG > MPPG > MSPG > MCPG. To date, MPPG (KD 9.2 microM) is the most potent L-AP4-sensitive receptor antagonist yet tested on the neonatal rat spinal cord. In addition, MTPG (KD 77 microM) is the most potent antagonist yet tested for (1S,3S)-ACPD-sensitive receptors in this preparation.
Assuntos
Antagonistas de Aminoácidos Excitatórios/síntese química , Antagonistas de Aminoácidos Excitatórios/farmacologia , Glicina/análogos & derivados , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Receptores Pré-Sinápticos/antagonistas & inibidores , Medula Espinal/efeitos dos fármacos , Aminobutiratos/farmacologia , Animais , Animais Recém-Nascidos , Cicloleucina/análogos & derivados , Cicloleucina/farmacologia , Glicina/síntese química , Glicina/farmacologia , Neurônios Motores/efeitos dos fármacos , Ratos , Tetrodotoxina/farmacologiaRESUMO
This study examined the binding of (S)-[3H]AMPA, the radiolabelled active isomer of AMPA, to rat brain synaptic membranes. Under non-chaotropic conditions specific binding of 10 nM (S)-[3H]AMPA represented 33 +/- 2% of the total; this increased to 74 +/- 1% in the presence of 100 mM KSCN. (S)-[3H]AMPA binding was inhibited by non-NMDA receptor agonists and the antagonists NBQX and CNQX, with the following rank order of potency: NBQX > (S)-AMPA > or = quisqualate > CNQX > L-glutamate > domoate > or = kainate > (R)-AMPA. NMDA, and the metabotropic glutamate receptor agonist (1S,3R)-ACPD, up to 100 microM, did not inhibit (S)-[3H]AMPA binding. A number of willardiine analogues all effectively inhibited (S)-[3H]AMPA binding with the rank order of potency: (S)-5-fluorowillardiine > (S)-5-nitrowillardiine > (S)-5-trifluoromethylwillardiine > (S)-5-bromowillardiine approximately (S)-5-chlorowillardiine > (S)-5-cyanowillardiine > (S)-willardiine > (S)-5-iodowillardiine > (S)-6-methylwillardiine > (S)-5-methylwillardiine. This rank order closely reflects data from equilibrium measurements made, under voltage clamp, on cultured hippocampal neurons. In contrast the respective (R)-enantiomers and the racemate mixtures of (R,S)-3, 5 and 6-isowillardiine were relatively inactive. Similar IC50 values and thus rank orders of potency for the willardiines were observed in the presence of 100 mM KSCN.
Assuntos
Alanina/análogos & derivados , Encéfalo/metabolismo , Membranas Sinápticas/metabolismo , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiônico/metabolismo , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Alanina/farmacologia , Animais , Encéfalo/ultraestrutura , Cicloleucina/análogos & derivados , Cicloleucina/farmacologia , Masculino , Pirimidinonas , Quinoxalinas/farmacologia , Ensaio Radioligante , Ratos , Ratos Wistar , Receptores de AMPA/efeitos dos fármacos , Estereoisomerismo , Trítio , UracilaRESUMO
Brief exposure of rat hippocampal slices to quisqualic acid (QUIS) sensitizes neurons to depolarization by the alpha-amino-omega-phosphonate excitatory amino acid (EAA) analogues AP4, AP5 and AP6. These phosphonates interact with a novel QUIS-sensitized site. Whereas L-AP4 and D-AP5 cross-react with other EAA receptors, DL-AP6 has been shown to be relatively selective for the QUIS-sensitized site. This specificity of DL-AP6, in conjunction with the apparent preference of this site for L-isomers, suggested that the hitherto unavailable L-isomer of AP6 would be a potent and specific agonist. We report the resolution of the D- and L-enantiomers of AP6 by fractional crystallization of the L-lysine salt of DL-AP6. We also report the pharmacological responses of kainate/AMPA, NMDA, lateral perforant path L-AP4 receptors and the CA1 QUIS-sensitized site to D- and L-AP6, and compare these responses to the D- and L-isomers of AP3, AP4, AP5 and AP7. The D-isomers of AP4, AP5 and AP6 were 5-, 3- and 14-fold less potent for the QUIS-sensitized site than their respective L-isomers. While L-AP4 and L-AP5 cross-reacted with NMDA and L-AP4 receptors, L-AP6 was found to be highly potent and specific for the QUIS-sensitized site (IC50 = 40 microM). Its IC50 values for kainate/AMPA, NMDA and L-AP4 receptors were > 10, 3 and 0.8 mM, respectively. As with AP4 and AP5, sensitization to L-AP6 was reversed by L-alpha-aminoadipate.