RESUMEN
The reproducibility of research using laboratory animals requires reliable management of their quality, in particular of their genetics, health and environment, all of which contribute to their phenotypes. The point at which these biological materials are transferred between researchers is particularly sensitive, as it may result in a loss of integrity of the animals and/or their documentation. Here, we describe the various aspects of laboratory animal quality that should be confirmed when sharing rodent research models. We also discuss how repositories of biological materials support the scientific community to ensure the continuity of the quality of laboratory animals. Both the concept of quality and the role of repositories themselves extend to all exchanges of biological materials and all networks that support the sharing of these reagents.
Asunto(s)
Investigadores , Animales , Humanos , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVES: Cell-based therapy has emerged as a promising strategy for the treatment of patients with heart failure. Increasing evidence supports the hypothesis that paracrine mechanisms mediated by soluble factors released by the cells play a predominate role in reparative processes. The aim of our study was to analyze which cytokines are released by CD34+ enriched cell products intended for autologous transendocardial CD34+ cell transplantation in patients with cardiomyopathy. MATERIAL AND METHODS: The peripheral blood CD34+ cells from 12 patients were mobilized with granulocyte colony-stimulating factor, collected via apheresis and enriched by immunoselection. RESULTS: In CD34+ enriched cell population, hematopoietic, but not mesenchymal or endothelial, progenitors were detected. Except for angiopoietin-1, other measured cytokines (FGF1, FGF2, VEGF, PDGF, IL-6, HGH, SDF-1α/CXCL12, NRG1) were not released by CD34+ cells. The average concentration of angiopoietin-1 released by 5×106 CD34+ cells grown in neutral DMEM medium was 213.6±130.0pg/mL (range: 74-448pg/mL). Angiopoietin-1 secretion correlated well with CD34+ cell's capacity for generating colonies derived from hematopoietic progenitors (Pearson's correlation=0.964; P<0.001). CONCLUSION: Our study presents angiopoietin-1 as an interesting candidate and suggests future studies to explore how its release by CD34+ cells might impact the success of autologous CD34+ cell transplantation.
Asunto(s)
Angiopoyetina 1/sangre , Antígenos CD34/análisis , Trasplante de Células Madre Hematopoyéticas , Células Madre Hematopoyéticas/metabolismo , Adulto , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Citocinas/análisis , Insuficiencia Cardíaca/etiología , Insuficiencia Cardíaca/terapia , Hemangioblastos/química , Hemangioblastos/citología , Hemangioblastos/metabolismo , Movilización de Célula Madre Hematopoyética , Células Madre Hematopoyéticas/química , Humanos , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/complicaciones , Neovascularización Fisiológica , Trasplante AutólogoRESUMEN
Dietary intake of methyl donors, such as folic acid and methionine, shows considerable intra-individual variation in human populations. While it is recognized that maternal departures from the optimum of dietary methyl donor intake can increase the risk for mental health issues and neurological disorders in offspring, it has not been explored whether paternal dietary methyl donor intake influences behavioral and cognitive functions in the next generation. Here, we report that elevated paternal dietary methyl donor intake in a mouse model, transiently applied prior to mating, resulted in offspring animals (methyl donor-rich diet (MD) F1 mice) with deficits in hippocampus-dependent learning and memory, impaired hippocampal synaptic plasticity and reduced hippocampal theta oscillations. Gene expression analyses revealed altered expression of the methionine adenosyltransferase Mat2a and BK channel subunit Kcnmb2, which was associated with changes in Kcnmb2 promoter methylation in MD F1 mice. Hippocampal overexpression of Kcnmb2 in MD F1 mice ameliorated altered spatial learning and memory, supporting a role of this BK channel subunit in the MD F1 behavioral phenotype. Behavioral and gene expression changes did not extend into the F2 offspring generation. Together, our data indicate that paternal dietary factors influence cognitive and neural functions in the offspring generation.
Asunto(s)
Cognición/fisiología , Suplementos Dietéticos/efectos adversos , Herencia Paterna/fisiología , Animales , Metilación de ADN , Dieta , Epigénesis Genética , Padre , Ácido Fólico/metabolismo , Hipocampo/metabolismo , Subunidades beta de los Canales de Potasio de Gran Conductancia Activados por el Calcio , Aprendizaje/efectos de los fármacos , Masculino , Memoria/efectos de los fármacos , Metionina/metabolismo , Metionina Adenosiltransferasa , Metilación , Ratones , Ratones Endogámicos C57BL , Neuronas/fisiología , Herencia Paterna/genética , Regiones Promotoras GenéticasRESUMEN
BACKGROUND/OBJECTIVES: Detailed protocols and recommendations for the assessment of energy balance have been provided to address the problems associated with different body mass and body composition as apparent for mouse models in obesity research. Here, we applied these guidelines to investigate energy balance in two inbred mouse strains with contrasting susceptibilities for diet-induced obesity (DIO). Mice of the AKR/J strain are highly susceptible, whereas the SWR/J mice are almost completely resistant. The proximate mechanisms responsible for this striking phenotypic difference are only partially understood. SUBJECTS/METHODS: Body mass and body composition, metabolizable energy, energy expenditure (EE), body temperature and spontaneous physical activity behavior were first assessed in a cohort of male AKR/J (N=29) and SWR/J (N=30) mice fed on a low-fat control diet (CD) to identify metabolic adaptations determining resistance to DIO. Thereafter, the immediate metabolic responses to high-fat diet (HFD) feeding for 3 days were investigated. Groups of weight-matched AKR/J (N=8) and SWR/J (N=8) mice were selected from the initial cohort for this intervention. RESULTS: Strain differences in body mass, fat mass and lean mass were adjusted by body mass as this was the only covariate significantly correlated with metabolizable energy and EE. On the CD, EE and fat oxidation was higher in SWR/J than in AKR/J mice, whereas no difference was found for metabolizable energy. In response to HFD feeding, both strains increased metabolizable energy intake, but also increased EE, body temperature, and fat oxidation. The catabolic adaptations to HFD feeding opposed the development of positive energy balance. Increased EE was not due to increased spontaneous physical activity. A significant strain difference was found when balancing metabolizable energy and daily energy expenditure (DEE). CONCLUSIONS: The guidelines were applicable with some limitations related to the adjustment of differences in body composition. Metabolic phenotyping revealed that metabolizable energy, DEE and metabolic fuel selection all contribute to the development of DIO. Therefore, assessing both sides of the energy balance equation is essential to identify the proximate mechanisms.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Obesidad/fisiopatología , Animales , Composición Corporal , Índice de Masa Corporal , Peso Corporal , Calorimetría Indirecta , Grasas de la Dieta , Metabolismo Energético , Ratones , Ratones Endogámicos AKR , Ratones Endogámicos , Obesidad/etiología , Condicionamiento Físico AnimalRESUMEN
OBJECTIVE: Excess lipid intake has been implicated in the pathophysiology of hepatosteatosis and hepatic insulin resistance. Lipids constitute approximately 50% of the cell membrane mass, define membrane properties, and create microenvironments for membrane-proteins. In this study we aimed to resolve temporal alterations in membrane metabolite and protein signatures during high-fat diet (HF)-mediated development of hepatic insulin resistance. METHODS: We induced hepatosteatosis by feeding C3HeB/FeJ male mice an HF enriched with long-chain polyunsaturated C18:2n6 fatty acids for 7, 14, or 21 days. Longitudinal changes in hepatic insulin sensitivity were assessed via the euglycemic-hyperinsulinemic clamp, in membrane lipids via t-metabolomics- and membrane proteins via quantitative proteomics-analyses, and in hepatocyte morphology via electron microscopy. Data were compared to those of age- and litter-matched controls maintained on a low-fat diet. RESULTS: Excess long-chain polyunsaturated C18:2n6 intake for 7 days did not compromise hepatic insulin sensitivity, however, induced hepatosteatosis and modified major membrane lipid constituent signatures in liver, e.g. increased total unsaturated, long-chain fatty acid-containing acyl-carnitine or membrane-associated diacylglycerol moieties and decreased total short-chain acyl-carnitines, glycerophosphocholines, lysophosphatidylcholines, or sphingolipids. Hepatic insulin sensitivity tended to decrease within 14 days HF-exposure. Overt hepatic insulin resistance developed until day 21 of HF-intervention and was accompanied by morphological mitochondrial abnormalities and indications for oxidative stress in liver. HF-feeding progressively decreased the abundance of protein-components of all mitochondrial respiratory chain complexes, inner and outer mitochondrial membrane substrate transporters independent from the hepatocellular mitochondrial volume in liver. CONCLUSIONS: We assume HF-induced modifications in membrane lipid- and protein-signatures prior to and during changes in hepatic insulin action in liver alter membrane properties - in particular those of mitochondria which are highly abundant in hepatocytes. In turn, a progressive decrease in the abundance of mitochondrial membrane proteins throughout HF-exposure likely impacts on mitochondrial energy metabolism, substrate exchange across mitochondrial membranes, contributes to oxidative stress, mitochondrial damage, and the development of insulin resistance in liver.
RESUMEN
Sequences encoding DUF1220 protein domains show the most extreme human lineage-specific copy number increase of any coding region in the genome and have been linked to human brain evolution. In addition, DUF1220 copy number (dosage) has been implicated in influencing brain size within the human species, both in normal populations and in individuals associated with brain size pathologies (1q21-associated microcephaly and macrocephaly). More recently, increasing dosage of a subtype of DUF1220 has been linked with increasing severity of the primary symptoms of autism. Despite these intriguing associations, a function for these domains has not been described. As a first step in addressing this question, we have developed the first transgenic model of DUF1220 function by removing the single DUF1220 domain (the ancestral form) encoded in the mouse genome. In a hypothesis generating exercise, these mice were evaluated by 197 different phenotype measurements. While resulting DUF1220-minus (KO) mice show no obvious anatomical peculiarities, they exhibit a significantly reduced fecundity (χ(2) = 19.1, df = 2, p = 7.0 × 10(-5)). Further extensive phenotypic analyses suggest hyperactivity (p < 0.05) of DUF1220 mice and changes in gene expression levels of brain associated with distinct neurological functions and disease. Other changes that met statistical significance include an increase in plasma glucose concentration (as measured by area under the curve, AUC 0-30 and AUC 30-120) in male mutants, fasting glucose levels, reduce sodium levels in male mutants, increased levels of the liver functional indicator ALAT/GPT in males, levels of alkaline phosphatase (also an indicator of liver function), mean R and SR amplitude by electrocardiography, elevated IgG3 levels, a reduced ratio of CD4:CD8 cells, and a reduced frequency of T cells; though it should be noted that many of these differences are quite small and require further examination. The linking of DUF1220 loss to a hyperactive phenotype is consistent with separate findings in which DUF1220 over expression results in a down-regulation of mitochondrial function, and potentially suggests a role in developmental metabolism. Finally, the substantially reduced fecundity we observe associated with KO mice argues that the ancestral DUF1220 domain provides an important biological functionthat is critical to survivability and reproductive success.
Asunto(s)
Evolución Biológica , Encéfalo/crecimiento & desarrollo , Fertilidad/genética , Dosificación de Gen , Ratones Transgénicos/genética , Fenotipo , Animales , Área Bajo la Curva , Glucemia/metabolismo , Calorimetría Indirecta , Cartilla de ADN/genética , Perfilación de la Expresión Génica , Técnicas de Inactivación de Genes , Hipercinesia/genética , Hígado/metabolismo , Masculino , Ratones , Tamaño de los Órganos , Estructura Terciaria de ProteínaRESUMEN
Breath gas analysis in humans proved successful in identifying disease states and assessing metabolic functions in a non-invasive way. While many studies report diagnostic capability using volatile organic compounds (VOC) in breath, the inter-individual variability even in healthy human cohorts is rather large and not completely understood in its biochemical origin. Laboratory mice are the predominant animal model system for human disorders and are analysed under highly standardized and controlled conditions. We established a novel setup to monitor VOCs as biomarkers for disease in the breath gas of non-anesthetized, non-restrained mice using a proton transfer reaction mass spectrometer with time of flight detection. In this study, we implemented breath gas analysis in a dietary intervention study in C57BL/6J mice with the aim to assess the variability in VOC signatures due to a change in the diet matrix. Mice were fed a standard laboratory chow and then exposed to four semi-purified low- or high-fat diets for four weeks. Random forest (RF++) was used to identify VOCs that specifically respond to the diet matrix change. Interestingly, we found that the change from a chow diet to semi-purified diets resulted in a considerable drop of several VOC levels. Our results suggest that the diet matrix impacts VOC signatures and the underlying metabolic functions and may be one source of variability in exhaled volatiles.
Asunto(s)
Pruebas Respiratorias/métodos , Dieta , Compuestos Orgánicos Volátiles/análisis , Acetatos/análisis , Animales , Biomarcadores/análisis , Sistemas de Computación , Dimetilsulfóxido/análisis , Espiración/fisiología , Conducta Alimentaria , Humanos , Modelos Lineales , Masculino , Espectrometría de Masas , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Propionatos/análisis , Sulfonas/análisis , Aumento de PesoRESUMEN
Vulnerability of the fetus upon maternal obesity can potentially occur during all developmental phases. We aimed at elaborating longer-term health outcomes of fetal overnutrition during the earliest stages of development. We utilized Naval Medical Research Institute (NMRI) mice to induce pre-conceptional and gestational obesity and followed offspring outcomes in the absence of any postnatal obesogenic influences. Male adult offspring developed overweight, insulin resistance, hyperleptinemia, hyperuricemia and hepatic steatosis; all these features were not observed in females. Instead, they showed impaired fasting glucose and a reduced fat mass and adipocyte size. Influences of the interaction of maternal diet∗sex concerned offspring genes involved in fatty liver disease, lipid droplet size regulation and fat mass expansion. These data suggest that a peri-conceptional obesogenic exposure is sufficient to shape offspring gene expression patterns and health outcomes in a sex- and organ-specific manner, indicating varying developmental vulnerabilities between sexes towards metabolic disease in response to maternal overnutrition.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Susceptibilidad a Enfermedades/fisiopatología , Obesidad/fisiopatología , Adipocitos/metabolismo , Adipocitos/patología , Animales , Peso Corporal/fisiología , Tamaño de la Célula , Susceptibilidad a Enfermedades/etiología , Hígado Graso/etiología , Hígado Graso/fisiopatología , Femenino , Regulación del Desarrollo de la Expresión Génica , Prueba de Tolerancia a la Glucosa , Hiperuricemia/etiología , Hiperuricemia/fisiopatología , Resistencia a la Insulina/fisiología , Leptina/sangre , Masculino , Ratones Endogámicos , Obesidad/etiología , Obesidad/genética , Sobrepeso/etiología , Sobrepeso/fisiopatología , Embarazo , Efectos Tardíos de la Exposición Prenatal/etiología , Efectos Tardíos de la Exposición Prenatal/genética , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores Sexuales , Grasa Subcutánea/metabolismo , Factores de TiempoRESUMEN
Several genetic diseases are triggered by nonsense mutations leading to the formation of truncated and defective proteins. Aminoglycosides have the capability to mediate a bypass of stop mutations during translation thus resulting in a rescue of protein expression. So far no attention has been directed to obesity-associated stop mutations as targets for nonsense suppression. Herein, we focus on the characterization of the melanocortin-4-receptor (MC4R) nonsense allele W16X identified in obese subjects. Cell culture assays revealed a loss-of-function of Mc4r(X16) characterized by impaired surface expression and defect signaling. The aminoglycoside G-418 restored Mc4r(X16) function in vitro demonstrating that Mc4r(X16) is susceptible to nonsense suppression. For the evaluation of nonsense suppression in vivo, we generated a Mc4r(X16) knock-in mouse line by gene targeting. Mc4r(X16) knock-in mice developed hyperphagia, impaired glucose tolerance, severe obesity and an increased body length demonstrating that this new mouse model resembles typical characteristics of Mc4r deficiency. In a first therapeutic trial, the aminoglycosides gentamicin and amikacin induced no amelioration of obesity. Further experiments with Mc4r(X16) knock-in mice will be instrumental to establish nonsense suppression for Mc4r as an obesity-associated target gene expressed in the central nervous system.
Asunto(s)
Codón sin Sentido , Receptor de Melanocortina Tipo 4/genética , Receptor de Melanocortina Tipo 4/metabolismo , Aminoglicósidos/genética , Aminoglicósidos/metabolismo , Animales , Composición Corporal/genética , Temperatura Corporal/genética , Peso Corporal/genética , Células COS , Línea Celular , Chlorocebus aethiops , Ingestión de Energía/genética , Expresión Génica/genética , Células HEK293 , Humanos , Hipotálamo/metabolismo , Ratones , Ratones Endogámicos C57BL , Obesidad/genética , Obesidad/metabolismoRESUMEN
Team of authors consisting of vitreoretinal surgeons and biomedical engineers developed and tested an electronic system helping the patients after some vitreoretinal surgeries to keep the recommended head position. The authors describe the principle of this system and its use in clinical practice.
Asunto(s)
Movimientos de la Cabeza , Monitoreo Ambulatorio/instrumentación , Cuidados Posoperatorios/instrumentación , Cirugía Vitreorretiniana , Electrónica Médica , HumanosRESUMEN
The Djungarian hamster is an animal that is prominent for distinct seasonal adaptations. Cued by shortening day length in autumn they spontaneously exhibit reductions in food intake, body mass (BM), fat mass and also in lean mass (LM). The mechanisms behind the seasonal regulation of body composition are only partly resolved. Although most studies focused on the participation of body fat in seasonal body weight regulation, we addressed the influence of LM, moreover of muscle mass (MM) on seasonal BM changes. Therefore, we analyzed body composition, MM and the expression of myostatin, a hormone negatively regulating muscle growth and differentiation, in Djungarian hamsters in response to naturally changing photoperiod in winter compared to long photoperiod (LP). Winter-acclimated hamsters upregulated myostatin mRNA when compared with hamsters adapted to natural and artificial LP, whereas MM remained unchanged when compared with natural LP. Moreover, in natural short photoperiod, individual myostatin expression levels were negatively correlated with MM. These results suggest that myostatin is under seasonal control in order to regulate MM and hence contributes to the overall LM and therefore BM changes in seasonal mammals.
Asunto(s)
Aclimatación/fisiología , Composición Corporal/fisiología , Músculo Esquelético/fisiología , Miostatina/fisiología , Phodopus/fisiología , Absorciometría de Fotón , Animales , Cricetinae , Miostatina/genética , Fotoperiodo , ARN/química , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estaciones del Año , Estadísticas no ParamétricasRESUMEN
The German Mouse Clinic (GMC) is a large scale phenotyping center where mouse mutant lines are analyzed in a standardized and comprehensive way. The result is an almost complete picture of the phenotype of a mouse mutant line--a systemic view. At the GMC, expert scientists from various fields of mouse research work in close cooperation with clinicians side by side at one location. The phenotype screens comprise the following areas: allergy, behavior, clinical chemistry, cardiovascular analyses, dysmorphology, bone and cartilage, energy metabolism, eye and vision, host-pathogen interactions, immunology, lung function, molecular phenotyping, neurology, nociception, steroid metabolism, and pathology. The German Mouse Clinic is an open access platform that offers a collaboration-based phenotyping to the scientific community (www.mouseclinic.de). More than 80 mutant lines have been analyzed in a primary screen for 320 parameters, and for 95% of the mutant lines we have found new or additional phenotypes that were not associated with the mouse line before. Our data contributed to the association of mutant mouse lines to the corresponding human disease. In addition, the systemic phenotype analysis accounts for pleiotropic gene functions and refines previous phenotypic characterizations. This is an important basis for the analysis of underlying disease mechanisms. We are currently setting up a platform that will include environmental challenge tests to decipher genome-environmental interactions in the areas nutrition, exercise, air, stress and infection with different standardized experiments. This will help us to identify genetic predispositions as susceptibility factors for environmental influences.
Asunto(s)
Investigación Biomédica/métodos , Modelos Animales de Enfermedad , Ratones Mutantes/genética , Fenotipo , Crianza de Animales Domésticos , Animales , Investigación Biomédica/normas , Alemania , Ratones , Ratones Mutantes/crecimiento & desarrollo , Control de CalidadRESUMEN
Obesity is associated with increased risk for developing pancreatic cancer, and it is suggested that insulin resistance provides the missing link. Here we demonstrate that under the context of genetic susceptibility, a high fat diet (HFD) predisposes mice with oncogenic K-ras activation to accelerated pancreatic intraepithelial neoplasm (PanIN) development. Tumor promotion is closely associated with increased inflammation and abrogation of TNFR1 signaling significantly blocks this process underlining a central role for TNFalpha in obesity-mediated enhancement of PanIN lesions. Interestingly, however, despite increased TNFalpha levels, mice remain insulin sensitive. We show that, while aggravating tumor promotion, a HFD exerts dramatic changes in energy metabolism through enhancement of pancreatic exocrine insufficiency, metabolic rates, and expression of genes involved in mitochondrial fatty acid (FA) beta-oxidation that collectively contribute to improved glucose tolerance in these mice. While on one hand these findings provide significant evidence that obesity is linked to tumor promotion in the pancreas, on the other it suggests alterations in inflammatory responses and bioenergetic pathways as the potential underlying cause.
Asunto(s)
Ácidos Grasos/metabolismo , Mitocondrias/metabolismo , Obesidad/metabolismo , Neoplasias Pancreáticas/metabolismo , Alimentación Animal , Animales , Progresión de la Enfermedad , Insuficiencia Pancreática Exocrina/inducido químicamente , Insuficiencia Pancreática Exocrina/metabolismo , Insuficiencia Pancreática Exocrina/patología , Ácidos Grasos/farmacología , Inflamación/genética , Inflamación/metabolismo , Resistencia a la Insulina , Ratones , Mitocondrias/efectos de los fármacos , Oxidación-Reducción , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patología , Receptores Tipo I de Factores de Necrosis Tumoral/metabolismo , Factores de Tiempo , Proteínas ras/metabolismoRESUMEN
Parkinson's disease (PD) is a progressive degenerative disorder of the central nervous system with tremor being one of its four main clinical features. Currently used methods can directly evaluate tremor amplitude and frequency but not joint movement in the affected limb. Measurement of joint movement facilitates the location of muscle groups that participate in PD tremor and this is important for treatment with local botulinum toxin injections. We developed and tested a method that measured tremor amplitude and frequency in a specific joint of the hand in PD patients. The tremor analysis method was based on force transducers adapted to record rest tremor of the wrist and metacarpophalangeal joints in two degrees of freedom for each joint. Direct measurements of joint movement in the hand can evaluate tremor amplitude and frequency and also locate the muscle groups that are most active in tremor movement, thus enabling their local treatment.
Asunto(s)
Técnicas Biosensibles/métodos , Mano/fisiopatología , Articulaciones/fisiopatología , Enfermedad de Parkinson/fisiopatología , Temblor/fisiopatología , Anciano , Fenómenos Biomecánicos , Técnicas Biosensibles/instrumentación , Humanos , Enfermedad de Parkinson/complicaciones , Temblor/diagnóstico , Temblor/etiologíaRESUMEN
A remarkable feature of the seasonal adaptation displayed by the Siberian hamster (Phodopus sungorus) is the ability to decrease food intake and body weight (by up to 40%) in response to shortening photoperiod. The regulating neuroendocrine systems involved in this adaptation and their neuroanatomical and molecular bases are poorly understood. We investigated the effect of photoperiod on the expression of prohormone convertases 1 (PC1/3) and 2 (PC2) and the endoproteolytic processing of the neuropeptide precursor pro-opiomelanocortin (POMC) within key energy balance regulating centres of the hypothalamus. We compared mRNA levels and protein distribution of PC1/3, PC2, POMC, adrenocorticotrophic hormone (ACTH), alpha-melanocyte-stimulating hormone (MSH), beta-endorphin and orexin-A in selected hypothalamic areas of long day (LD, 16:8 h light:dark), short day (SD, 8:16 h light:dark) and natural-day (ND, photoperiod depending on time of the year) acclimated Siberian hamsters. The gene expression of PC2 was significantly higher within the arcuate nucleus (ARC, P < 0.01) in SD and in ND (versus LD), and is reflected in the day length profile between October and April in the latter. PC1/3 gene expression in the ARC and lateral hypothalamus was higher in ND but not in SD compared to the respective LD controls. The immunoreactivity of PC1/3 cleaved neuropeptide ACTH in the ARC and PC1/3-colocalised orexin-A in the lateral hypothalamus were not affected by photoperiod changes. However, increased levels of PC2 mRNA and protein were associated with higher abundance of the mature neuropeptides alpha-MSH and beta-endorphin (P < 0.01) in SD. This study provides a possible explanation for previous paradoxical findings showing lower food intake in SD associated with decreased POMC mRNA levels. Our results suggest that a major part of neuroendocrine body weight control in seasonal adaptation may be effected by post-translational processing mediated by the prohormone convertases PC1/3 and PC2, in addition to regulation of gene expression of neuropeptide precursors.
Asunto(s)
Adaptación Fisiológica/genética , Fotoperiodo , Proopiomelanocortina/genética , Proproteína Convertasa 1/genética , Proproteína Convertasa 2/genética , Hormona Adrenocorticotrópica/metabolismo , Animales , Peso Corporal/fisiología , Cricetinae , Femenino , Regulación Enzimológica de la Expresión Génica/fisiología , Área Hipotalámica Lateral/fisiología , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Neuropéptidos/genética , Neuropéptidos/metabolismo , Orexinas , Phodopus , Proopiomelanocortina/metabolismo , Proproteína Convertasa 1/metabolismo , Proproteína Convertasa 2/metabolismo , Precursores de Proteínas/genética , Procesamiento Proteico-Postraduccional/fisiología , ARN Mensajero/análisis , Estaciones del Año , alfa-MSH/metabolismo , betaendorfina/metabolismoRESUMEN
Postural instability has a big impact on the quality of life of patients with Parkinson's disease (PD) as it often leads to an insecure stance and fall. We investigated if postural stability in these patients improves by decreasing rigidity with a dopaminergic agonist. In our study, we tested eight PD patients with no concomitant diseases. Their age was 61 +/- 2 years (mean +/- SE) and their Hoehn-Yahr score was 3 +/- 0.1. The patients were evaluated according to the Unified Parkinson's Disease Rating Scale for motor function (mUPDRS) and with stabilometric measurements of forward-backward and side-to-side body oscillations during free stance with eyes open. Both evaluations were performed in an "off "state and in an apomorphine-induced "on" state. As expected, the mUPDRS score was significantly decreased in the "on" state with posture being improved in six patients, gait in eight patients and postural stability in seven of eight patients. In addition, apomorphine caused a significant reduction of the relative amplitude of lower frequencies and an increase of the relative amplitude of higher frequencies of forward-backward body oscillations. The results of stabilometry and mUPDRS evaluations are in agreement with the effect of apomorphine on rigidity, indicating that postural stability of PD patients is improved by decreasing rigidity.
Asunto(s)
Rigidez Muscular/fisiopatología , Enfermedad de Parkinson/fisiopatología , Postura/fisiología , Antiparkinsonianos/uso terapéutico , Apomorfina/uso terapéutico , Humanos , Persona de Mediana Edad , Rigidez Muscular/tratamiento farmacológico , Enfermedad de Parkinson/tratamiento farmacológicoRESUMEN
The superficial regions of the left vagus nerves of a dog were selectively stimulated with 39-electrode spiral cuffs having 13 circumferential groups of three electrodes (GTE) to modulate the function of the innervated internal organs and glands. Under general anaesthesia, the cuffs were chronically implanted around the nerve in the neck in two adult Beagle dogs and remained viable for 16 months. The regions were stimulated with biphasic, rectangular current pulses (2 mA, 200 micros, 20 Hz) delivered to the group of GTE lying close to the region innervating the specific internal organs or glands. The results showed that specific electrode configurations had actions on the heart (GTE 9), lungs (GTE 4) and pressure in the urinary bladder (GTE 1). It was also shown that GTE no. 10 significantly modified the endocrine function of the pancreas. The results of this study clearly demonstrate that internal organs and glands can be selectively stimulated via the selective stimulation of innervating superficial regions of the autonomous peripheral nerve.
Asunto(s)
Corazón/fisiología , Islotes Pancreáticos/fisiología , Pulmón/fisiología , Vejiga Urinaria/fisiología , Nervio Vago/fisiología , Animales , Perros , Estimulación Eléctrica/instrumentación , Electrodos Implantados , Diseño de Equipo , PresiónRESUMEN
The purpose of the present study was to modulate the secretion of insulin and glucagon in Beagle dogs by stimulation of nerves innervating the intact and partly dysfunctional pancreas. Three 33-electrode spiral cuffs were implanted on the vagus, splanchnic and pancreatic nerves in each of two animals. Partial dysfunction of the pancreas was induced with alloxan. The nerves were stimulated using rectangular, charge-balanced, biphasic, and constant current pulses (200 micros, 1 mA, 20 Hz, with a 100-micros delay between biphasic phases). Blood samples from the femoral artery were drawn before the experiment, at the beginning of stimulation, after 5 min of stimulation, and 5 min after the end of stimulation. Radioimmunoassay data showed that in the intact pancreas stimulation of the vagal nerve increased insulin (+99.2 microU/ml) and glucagon (+18.7 pg/ml) secretion and decreased C-peptide secretion (-0.15 ng/ml). Splanchnic nerve stimulation increased insulin (+1.7 microU/ml), C-peptide (+0.01 ng/ml), and glucagon (+50 pg/ml) secretion, whereas pancreatic nerve stimulation did not cause a marked change in any of the three hormones. In the partly dysfunctional pancreas, vagus nerve stimulation increased insulin (+15.5 microU/ml), glucagon (+11 pg/ml), and C-peptide (+0.03 ng/ml) secretion. Splanchnic nerve stimulation reduced insulin secretion (-2.5 microU/ml) and increased glucagon (+58.7 pg/ml) and C-peptide (+0.39 ng/ml) secretion, and pancreatic nerve stimulation increased insulin (+0.2 microU/ml), glucagon (+5.2 pg/ml), and C-peptide (+0.08 ng/ml) secretion. It was concluded that vagal nerve stimulation can significantly increase insulin secretion for a prolonged period of time in intact and in partly dysfunctional pancreas.
Asunto(s)
Estimulación Eléctrica/métodos , Glucagón/metabolismo , Insulina/metabolismo , Páncreas/metabolismo , Animales , Diabetes Mellitus Experimental/inducido químicamente , Diabetes Mellitus Experimental/metabolismo , Perros , Electrodos Implantados , Secreción de Insulina , Páncreas/inervación , Radioinmunoensayo , Nervios Esplácnicos/fisiología , Nervio Vago/fisiologíaRESUMEN
The purpose of the present study was to modulate the secretion of insulin and glucagon in Beagle dogs by stimulation of nerves innervating the intact and partly dysfunctional pancreas. Three 33-electrode spiral cuffs were implanted on the vagus, splanchnic and pancreatic nerves in each of two animals. Partial dysfunction of the pancreas was induced with alloxan. The nerves were stimulated using rectangular, charge-balanced, biphasic, and constant current pulses (200 µs, 1 mA, 20 Hz, with a 100-µs delay between biphasic phases). Blood samples from the femoral artery were drawn before the experiment, at the beginning of stimulation, after 5 min of stimulation, and 5 min after the end of stimulation. Radioimmunoassay data showed that in the intact pancreas stimulation of the vagal nerve increased insulin (+99.2 µU/ml) and glucagon (+18.7 pg/ml) secretion and decreased C-peptide secretion (-0.15 ng/ml). Splanchnic nerve stimulation increased insulin (+1.7 µU/ml), C-peptide (+0.01 ng/ml), and glucagon (+50 pg/ml) secretion, whereas pancreatic nerve stimulation did not cause a marked change in any of the three hormones. In the partly dysfunctional pancreas, vagus nerve stimulation increased insulin (+15.5 µU/ml), glucagon (+11 pg/ml), and C-peptide (+0.03 ng/ml) secretion. Splanchnic nerve stimulation reduced insulin secretion (-2.5 µU/ml) and increased glucagon (+58.7 pg/ml) and C-peptide (+0.39 ng/ml) secretion, and pancreatic nerve stimulation increased insulin (+0.2 µU/ml), glucagon (+5.2 pg/ml), and C-peptide (+0.08 ng/ml) secretion. It was concluded that vagal nerve stimulation can significantly increase insulin secretion for a prolonged period of time in intact and in partly dysfunctional pancreas.
Asunto(s)
Animales , Perros , Estimulación Eléctrica , Glucagón , Insulina , Páncreas , Electrodos Implantados , Glucagón , Insulina , Páncreas , Radioinmunoensayo , Nervios Esplácnicos , Nervio VagoRESUMEN
Electroneurograms (ENGs) from the vagus, splanchnic and pancreatic nerves innervating the pancreas of a dog, were recorded with chronically implanted silicone multi-electrode circular cuffs in an intact pancreas and in a pancreas partly disabled with alloxan. The cuffs contained 33 platinum electrodes (0.6x1.5 mm) arranged in three parallel circular groups integrated into the inner surface of the cuff. Each circular group contained 11 electrodes at a distance of 0.5 mm apart, with 6 mm between the circular groups. The cuffs had an inner diameter of 2.5 mm and the length of 18 mm. In a 2-year study, the cuffs were implanted into two adult Beagle dogs (one female and one male). In the vagus nerve, the cuff was installed on the nerve at the neck, whilst in the splanchnic nerve, the cuff was installed on the nerve before the celiac ganglion, and in the pancreatic nerve, the cuff was installed on the nerve just before it enters the pancreas. In each of the three implanted cuffs, the electrodes of the central circular group were connected to each other and this signal provided one input to a multi-channel ENG amplifying system. The electrodes of each of the two outer spiral groups were connected to each other and then both these groups were short-circuited. This signal then provided another input to the multi-channel ENG amplifying system. The ENG amplifying system was designed to amplify the ENGs 100000 times and to pass frequencies of between 500 and 10 kHz. In our study, three recordings in each animal were conducted. Recordings in the intact pancreas were conducted 2 and 6 months after the implantation, while the recording in the partly disabled pancreas, was conducted 10 months after the implantation and 10 days after the disablement. Due to the fact that the results obtained in both animals were actually quite similar, we present the results of the recordings obtained in one animal. In both animals the cuffs were left implanted for more than 1 year and were used for pancreatic stimulation, although this is not in this paper. The results show that cuffs implanted chronically on the nerves innervating the pancreas of a dog could reliably record the ENGs. This information could be used effectively in further study of pancreatic innervation and its function. Moreover, the results suggest that cuffs could also be useful in recording the ENGs from other nerves of the autonomic nervous system that innervate various glands and internal organs.