RESUMEN
Here we show that central administration of pyroglutamylated arginine-phenylamine-amide peptide (QRFP/26RFa) increases both food intake and locomotor activity, without any significant effect on energy expenditure, thermogenesis or reward. Germline knock out of either of the mouse QRFP receptor orthologs, Gpr103a and Gpr103b, did not produce a metabolic phenotype. However, both receptors are required for the effect of centrally administered QRFP to increase feeding and locomotor activity. As central injection of QRFP activated orexin/hypocretin neurons in the lateral hypothalamus, we compared the action of QRFP and orexin on behaviour. Both peptides increased arousal and locomotor activity. However, while orexin increased consummatory behaviour, QRFP also affected other appetitive behaviours. Furthermore, the feeding but not the locomotor response to QRFP, was blocked by co-administration of an orexin receptor 1 antagonist. These results suggest that QRFP agonism induces both appetitive and consummatory behaviour, but only the latter is dependent on orexin/hypocretin receptor signalling.
Asunto(s)
Receptores de Orexina , Péptidos , Receptores Acoplados a Proteínas G , Animales , Ratones , Amidas , Compuestos de Anilina , Arginina , Péptidos y Proteínas de Señalización Intercelular , Locomoción , Neuropéptidos , Receptores de Orexina/metabolismo , Orexinas , Péptidos/farmacología , Receptores Acoplados a Proteínas G/metabolismo , Conducta AlimentariaRESUMEN
Growth/differentiation factor 15 (GDF15), also known as MIC-1, is a distant member of the transforming growth factor-ß (TGF-ß) superfamily and has been implicated in various biological functions, including cancer cachexia, renal and heart failure, atherosclerosis and metabolism. A connection between GDF15 and body-weight regulation was initially suggested on the basis of an observation that increasing GDF15 levels in serum correlated with weight loss in individuals with advanced prostate cancer. In animal models, overexpression of GDF15 leads to a lean phenotype, hypophagia and other improvements in metabolic parameters, suggesting that recombinant GDF15 protein could potentially be used in the treatment of obesity and type 2 diabetes. However, the signaling and mechanism of action of GDF15 are poorly understood owing to the absence of a clearly identified cognate receptor. Here we report that GDNF-family receptor α-like (GFRAL), an orphan member of the GFR-α family, is a high-affinity receptor for GDF15. GFRAL binds to GDF15 in vitro and is required for the metabolic actions of GDF15 with respect to body weight and food intake in vivo in mice. Gfral-/- mice were refractory to the effects of recombinant human GDF15 on body-weight, food-intake and glucose parameters. Blocking the interaction between GDF15 and GFRAL with a monoclonal antibody prevented the metabolic effects of GDF15 in rats. Gfral mRNA is highly expressed in the area postrema of mouse, rat and monkey, in accordance with previous reports implicating this region of the brain in the metabolic actions of GDF15 (refs. 4,5,6). Together, our data demonstrate that GFRAL is a receptor for GDF15 that mediates the metabolic effects of GDF15.
Asunto(s)
Área Postrema/metabolismo , Ingestión de Alimentos/efectos de los fármacos , Receptores del Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Factor 15 de Diferenciación de Crecimiento/farmacología , Obesidad/metabolismo , Pérdida de Peso/efectos de los fármacos , Animales , Encéfalo/metabolismo , Ingestión de Alimentos/genética , Citometría de Flujo , Receptores del Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Células HEK293 , Humanos , Immunoblotting , Macaca fascicularis , Masculino , Ratones , Ratones Noqueados , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Resonancia por Plasmón de Superficie , Pérdida de Peso/genéticaRESUMEN
STUDY DESIGN: Immunohistological study. OBJECTIVE: To elucidate the role of matrix metalloproteinases (MMPs), hypoxia-inducible factor-1α (HIF), and vascular endothelial growth factor (VEGF) in the hypertrophied ligamentum flavum (LF) obtained from patients with lumbar spinal stenosis (LSS). SUMMARY OF BACKGROUND DATA: The most common spinal disorder in the elderly is LSS, which results in part from LF hypertrophy. Although prior histologic and immunochemical studies have been performed in this area, the pathophysiology of loss of elasticity and hypertrophy is not completely understood. METHODS: LF samples of 38 patients with LSS were harvested during spinal decompression. Twelve LF samples obtained from patients with disk herniation and no visible degeneration on preoperative magnetic resonance imaging were obtained as controls. Samples were dehydrated and paraffin embedded. For immunohistochemical determination of VEGF, HIF, and MMPs 1, 3, and 9 expression, slices were stained with VEGF, HIF, and MMP antibody dilution. Neovessel density and number of elastic fibers were counted after Masson-Goldner staining. LF hypertrophy and cross-sectional area (CSA) were measured on T1-weighted magnetic resonance imaging. RESULTS: MMPs 1, 3, 9 and VEGF expression were significantly increased in the hypertrophy group (P<0.05). HIF expression was negative in both groups. Vessel density was increased in the hypertrophy group, although this was not statistically significant. The number of elastic fibres was significantly higher in the control group. In the hypertrophy group, LF thickness was significantly increased, whereas CSA was significantly decreased. There was a statistical correlation between LF thickness, CSA, MMP, and VEGF expression in the hypertrophy group (P<0.05). CONCLUSIONS: LF hypertrophy is accompanied by increased MMPs 1, 3, 9 and VEGF expression. Neovessel density is increased in hypertrophied LF. HIF is not expressed in hypertrophied LF.
Asunto(s)
Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Ligamento Amarillo/enzimología , Ligamento Amarillo/patología , Metaloproteinasas de la Matriz/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Demografía , Femenino , Humanos , Hipertrofia/patología , Desplazamiento del Disco Intervertebral , Ligamento Amarillo/diagnóstico por imagen , Vértebras Lumbares/enzimología , Vértebras Lumbares/patología , Imagen por Resonancia Magnética , Masculino , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Persona de Mediana Edad , Radiografía , Estenosis Espinal/enzimología , Estenosis Espinal/patología , Adulto JovenRESUMEN
BACKGROUND: The most common spinal disorder in the elderly is lumbar spinal stenosis (LSS), which results in part from ligamentum flavum (LF) hypertrophy. Although prior histologic and immunochemical studies have been performed in this area, the pathophysiology of loss of elasticity and hypertrophy is not completely understood. The purpose of this immunohistological study is to elucidate the role of CD44 and its splice variants CD44v5 and CD44v6 in the hypertrophied LF obtained from patients with lumbar spinal stenosis (LSS). MATERIALS AND METHODS: LF samples of 38 patients with LSS were harvested during spinal decompression. Twelve LF samples obtained from patients with disc herniation and no visible degeneration on preoperative MRI were obtained as controls. Samples were dehydrated and embedded in paraffin. For immunohistochemical determination, slices were stained with antibodies against CD44, Cd44v4, and CD44v6 stained with DAB. LF hypertrophy and cross-sectional area (CSA) were measured with T1-weighted MRI. RESULTS: CD44 and CD44v5 expression were significantly increased in the hypertrophy group (p < 0.05). CD44v6 expression was not significantly increased. The number of elastic fibers was significantly higher in the hypertrophy group. In the hypertrophy group, LF thickness was significantly increased while CSA was significantly decreased. There was a statistical correlation between LF thickness, CSA, CD44, and CD44v5 expression in the hypertrophy group (p < 0.05). CONCLUSIONS: LF hypertrophy is accompanied by increased CD44 and CD44v5 expression. CD44v6 expression is not enhanced in LF hypertrophy.
Asunto(s)
Receptores de Hialuranos/metabolismo , Desplazamiento del Disco Intervertebral/metabolismo , Desplazamiento del Disco Intervertebral/patología , Ligamento Amarillo/metabolismo , Ligamento Amarillo/patología , Anciano , Anciano de 80 o más Años , Biomarcadores/metabolismo , Femenino , Humanos , Hipertrofia/complicaciones , Hipertrofia/diagnóstico , Hipertrofia/metabolismo , Inmunohistoquímica , Desplazamiento del Disco Intervertebral/complicaciones , Vértebras Lumbares/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Radiografía , Estenosis Espinal/diagnóstico , Estenosis Espinal/etiologíaRESUMEN
Acute phencyclidine induces schizophrenia-like symptoms in healthy humans and psychotic episodes in schizophrenics. Although phencyclidine is known as a N-methyl d-aspartate receptor antagonist (NMDA-R), the molecular events underlying the behavioral symptoms remain largely unknown. Statistical analysis of oligonucleotide microarray data was used to identify phencyclidine-induced alterations in rat cortical gene expression. Acute phencyclidine produced a statistically significant change in 477 genes in rat prefrontal cortex (PFC), a brain area associated with cognitive dysfunction in schizophrenics. Real-time quantitative PCR (RTQ-PCR) confirmed a subset of these changes ranging from -59% to 255% (smallest confirmation: -19%). Subsequent time-course and dose-response studies using RTQ-PCR confirmed and extended the original microarray results. At the molecular level, genes altered by phencyclidine are related to diverse biological processes including stress, inflammatory response, growth and development, neural plasticity and signal transduction. Further analysis, aimed at assessing the relevance of our results to schizophrenia, revealed dysregulation of genes related to: (i) thalamocortical projections, (ii) neurotransmission and neuromodulation, (iii) thyroid hormone activity, (iv) oligodendrocyte linage, (v) brain lipid metabolism, (vi) sleep architecture and (viii) the velocardiofacial syndrome.