Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 9 de 9
Filtrar
1.
Neuron ; 89(4): 842-56, 2016 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-26889812

RESUMEN

Neuromodulation of arousal states ensures that an animal appropriately responds to its environment and engages in behaviors necessary for survival. However, the molecular and circuit properties underlying neuromodulation of arousal states such as sleep and wakefulness remain unclear. To tackle this challenge in a systematic and unbiased manner, we performed a genetic overexpression screen to identify genes that affect larval zebrafish arousal. We found that the neuropeptide neuromedin U (Nmu) promotes hyperactivity and inhibits sleep in zebrafish larvae, whereas nmu mutant animals are hypoactive. We show that Nmu-induced arousal requires Nmu receptor 2 and signaling via corticotropin releasing hormone (Crh) receptor 1. In contrast to previously proposed models, we find that Nmu does not promote arousal via the hypothalamic-pituitary-adrenal axis, but rather probably acts via brainstem crh-expressing neurons. These results reveal an unexpected functional and anatomical interface between the Nmu system and brainstem arousal systems that represents a novel wake-promoting pathway.


Asunto(s)
Regulación de la Expresión Génica/genética , Neuropéptidos/genética , Neuropéptidos/metabolismo , Sueño/genética , Vigilia/genética , Factores de Edad , Compuestos de Anilina/farmacología , Animales , Tronco Encefálico/citología , Tronco Encefálico/crecimiento & desarrollo , Tronco Encefálico/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Sistema Hipotálamo-Hipofisario/metabolismo , Larva , Ratones Transgénicos , Actividad Motora/genética , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Sistema Hipófiso-Suprarrenal/metabolismo , Pirimidinas/farmacología , Receptores de Complemento 3b/metabolismo , Receptores de Neurotransmisores/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Pez Cebra , Proteínas de Pez Cebra/genética
2.
Cereb Cortex ; 25(10): 3713-22, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25316336

RESUMEN

During cortical development, synaptic competition regulates the formation and adjustment of neuronal connectivity. It is unknown whether synaptic competition remains active in the adult brain and how inhibitory neurons participate in this process. Using morphological and electrophysiological measurements, we show that expressing a dominant-negative form of the TrkB receptor (TrkB.T1) in the majority of pyramidal neurons in the adult visual cortex does not affect excitatory synapse densities. This is in stark contrast to the previously reported loss of excitatory input which occurs if the exact same transgene is expressed in sparse neurons at the same age. This indicates that synaptic competition remains active in adulthood. Additionally, we show that interneurons not expressing the TrkB.T1 transgene may have a competitive advantage and obtain more excitatory synapses when most neighboring pyramidal neurons do express the transgene. Finally, we demonstrate that inhibitory synapses onto pyramidal neurons are reduced when TrkB signaling is interfered with in most pyramidal neurons but not when few pyramidal neurons have this deficit. This adjustment of inhibitory innervation is therefore not a cell-autonomous consequence of decreased TrkB signaling but more likely a homeostatic mechanism compensating for activity changes at the population level.


Asunto(s)
Potenciales Postsinápticos Excitadores , Potenciales Postsinápticos Inhibidores , Células Piramidales/fisiología , Receptor trkB/metabolismo , Corteza Visual/fisiología , Potenciales de Acción , Animales , Espinas Dendríticas/metabolismo , Homeostasis , Interneuronas/metabolismo , Interneuronas/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Potenciales Postsinápticos Miniatura , Células Piramidales/metabolismo , Receptor trkB/genética , Receptores AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Transducción de Señal , Sinapsis/metabolismo , Sinapsis/fisiología , Corteza Visual/metabolismo
3.
Zebrafish ; 11(2): 154-66, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24428353

RESUMEN

Compounds routinely used to increase the quality of life and combat disease undergo stringent potency and biosafety tests before approval. However, based on the outcome of ongoing research, new norms need to be effected to ensure that the compounds conform to biosafety at all target levels of activity. Whereas in vitro tests used to assess biosafety lack the potency and the translational attribute of a whole animal, mammalian preclinical models are expensive and time exhaustive. Zebrafish (Danio rerio) has emerged as an attractive alternative for biosafety studies due to its small size, genetics, breeding capabilities, and most importantly, similarity at the molecular and physiological levels with humans. It has been used extensively for testing various forms of toxicity, including developmental toxicity, cardiotoxicity, nephrotoxicity, and hepatotoxicity. We review here the utility of zebrafish as a powerful, sensitive, quantitative, noninvasive, and high-throughput whole-animal assay to screen for toxicity. Different forms of toxicity will be discussed briefly before we highlight the present state of genotoxicity study in zebrafish. This review, a first in this research area, will serve as a comprehensive introduction to the field of genotoxicity assay using zebrafish, a nascent but promising field that assays compounds for DNA damage. We also discuss possible approaches that could potentially be pursued to overcome some of the shortcomings in current genotoxic studies.


Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Modelos Animales , Pruebas de Mutagenicidad/métodos , Pez Cebra/genética , Animales
4.
Nat Neurosci ; 13(5): 642-8, 2010 May.
Artículo en Inglés | MEDLINE | ID: mdl-20400960

RESUMEN

During development and aging and in amblyopia, visual acuity is far below the limitations set by the retina. Expression of brain-derived neurotrophic factor (BDNF) in the visual cortex is reduced in these situations. We asked whether neurotrophic tyrosine kinase receptor, type 2 (TrkB) regulates cortical visual acuity in adult mice. We found that genetically interfering with TrkB/BDNF signaling in pyramidal cells in the mature visual cortex reduced synaptic strength and resulted in a loss of neural responses to high spatial-frequency stimuli. Responses to low spatial-frequency stimuli were unaffected. This selective loss was not accompanied by a change in receptive field sizes or plasticity, but apparent contrast was reduced. Our results indicate that a dependence on spatial frequency in the Heeger normalization model explains this selective effect of contrast reduction on high-resolution vision and suggest that it involves contrast gain control operating in the visual cortex.


Asunto(s)
Sensibilidad de Contraste/fisiología , Receptor trkB/metabolismo , Transducción de Señal/fisiología , Sinapsis/fisiología , Agudeza Visual/fisiología , Corteza Visual/metabolismo , Factores de Edad , Animales , Biofisica , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Predominio Ocular/fisiología , Estimulación Eléctrica/métodos , Proteínas Fluorescentes Verdes/genética , Técnicas In Vitro , Ratones , Ratones Transgénicos , Modelos Neurológicos , Inhibición Neural/fisiología , Plasticidad Neuronal/fisiología , Técnicas de Placa-Clamp/métodos , Estimulación Luminosa/métodos , Valor Predictivo de las Pruebas , Células Piramidales/metabolismo , Tiempo de Reacción/fisiología , Potenciales Sinápticos/genética , Corteza Visual/citología
5.
Behav Neurosci ; 123(6): 1197-217, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20001104

RESUMEN

We have demonstrated in our previous studies that ventral subicular lesion induces neurodegeneration of the hippocampus and produces cognitive impairment in rats. In the present study, the efficacy of transplanted green fluorescent protein (GFP)-labeled hippocampal cell line (H3-GFP) cells in establishing functional recovery in ventral subicular lesioned rats has been evaluated. The survival of H3-GFP transplants and their ability to express trophic factors in vivo were also investigated. Adult male Wistar rats were subjected to selective lesioning of ventral subiculum and were transplanted with H3-GFP cells into the cornu ammonis 1 (CA1) hippocampus. The transplants settled mainly in the dentate gyrus and expressed neurotrophic factors, brain-derived neurotrophic factor (BDNF), and basic fibroblast growth factor (bFGF). The ventral subicular lesioned (VSL) rats with H3-GFP transplants showed enhanced expression of BDNF in the hippocampus and performed well in eight-arm radial maze and Morris water maze tasks. The VSL rats without hippocampal transplants continued to show cognitive impairment in task learning. The present study demonstrated the H3-GFP transplants mediated recovery of cognitive functions in VSL rats. Our study supports the notion of graft meditated host regeneration and functional recovery through trophic support, although these mechanisms require further investigation.


Asunto(s)
Trasplante de Células , Hipocampo/citología , Hipocampo/metabolismo , Aprendizaje por Laberinto/fisiología , Análisis de Varianza , Animales , Conducta Animal/fisiología , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Recuento de Células , Línea Celular , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Técnica del Anticuerpo Fluorescente , Proteínas Fluorescentes Verdes/metabolismo , Hipocampo/patología , Procesamiento de Imagen Asistido por Computador , Masculino , Microscopía Confocal , Degeneración Nerviosa/metabolismo , Degeneración Nerviosa/patología , Neuronas/metabolismo , Neuronas/patología , Ratas , Ratas Wistar , Recuperación de la Función/fisiología , Conducta Espacial/fisiología , Factores de Tiempo
6.
Mol Cell Endocrinol ; 311(1-2): 77-86, 2009 Nov 13.
Artículo en Inglés | MEDLINE | ID: mdl-19643163

RESUMEN

'AT(4) receptors' through which Angiotensin IV (Ang IV) improves memory acquisition, were recently identified as insulin regulated aminopeptidase (IRAP). Radioligand binding studies have hitherto been performed with iodinated Ang IV in the presence of divalent cation chelators EDTA and 1,10-phenanthrolin. Hence, they referred to the apo-form of IRAP. Presently, binding of [(3)H]Ang IV and [(3)H]AL-11, a stable Ang IV analog, was compared on Chinese hamster ovary (CHO-K1) and mouse hippocampal (P40H1) cell membranes. With chelators, their high affinity sites showed the same pharmacological profile as for [(125)I]Ang IV binding. Without chelators, only high affinity binding was perceived for [(3)H]AL-11. The same pharmacological profile was recorded in both membrane preparations; it was different from the one in the presence of chelators and corresponded to catalytically active IRAP (despite the concurrent presence of aminopeptidase N (APN) in P40H1 cell membranes). This confirms that the active and apo-forms of IRAP have a distinct pharmacological profile.


Asunto(s)
Angiotensina II/análogos & derivados , Cistinil Aminopeptidasa/metabolismo , Receptores de Angiotensina/metabolismo , Coloración y Etiquetado , Angiotensina II/metabolismo , Animales , Unión Competitiva , Bioensayo , Células CHO , Cricetinae , Cricetulus , Cistinil Aminopeptidasa/antagonistas & inhibidores , Humanos , Cinética , Ligandos , Ratones , Tritio
7.
PLoS One ; 3(8): e3059, 2008 Aug 26.
Artículo en Inglés | MEDLINE | ID: mdl-18725976

RESUMEN

BACKGROUND: Transgenic mice with mosaic, Golgi-staining-like expression of enhanced green fluorescent protein (EGFP) have been very useful in studying the dynamics of neuronal structure and function. In order to further investigate the molecular events regulating structural plasticity, it would be useful to express multiple proteins in the same sparse neurons, allowing co-expression of functional proteins or co-labeling of subcellular compartments with other fluorescent proteins. However, it has been difficult to obtain reproducible expression in the same subset of neurons for direct comparison of neurons expressing different functional proteins. PRINCIPAL FINDINGS: Here we describe a Cre-transgenic line that allows reproducible expression of transgenic proteins of choice in a small number of neurons of the adult cortex, hippocampus, striatum, olfactory bulb, subiculum, hypothalamus, superior colliculus and amygdala. We show that using these Cre-transgenic mice, multiple Cre-dependent transgenes can be expressed together in the same isolated neurons. We also describe a Cre-dependent transgenic line expressing a membrane associated EGFP (EGFP-F). Crossed with the Cre-transgenic line, EGFP-F expression starts in the adolescent forebrain, is present in dendrites, dendritic protrusions, axons and boutons and is strong enough for acute or chronic in vivo imaging. SIGNIFICANCE: This triple transgenic approach will aid the morphological and functional characterization of neurons in various Cre-dependent transgenic mice.


Asunto(s)
Neuronas/fisiología , Prosencéfalo/fisiología , Adulto , Animales , Encéfalo/fisiología , Expresión Génica , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Humanos , Integrasas/genética , Integrasas/metabolismo , Ratones , Ratones Transgénicos , Mosaicismo
8.
Proc Natl Acad Sci U S A ; 103(4): 1071-6, 2006 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-16418274

RESUMEN

In adult primary visual cortex (V1), dendritic spines are more persistent than during development. Brain-derived neurotrophic factor (BDNF) increases synaptic strength, and its levels rise during cortical development. We therefore asked whether postsynaptic BDNF signaling through its receptor TrkB regulates spine persistence in adult V1. This question has been difficult to address because most methods used to alter TrkB signaling in vivo affect cortical development or cannot distinguish between pre- and postsynaptic mechanisms. We circumvented these problems by employing transgenic mice expressing a dominant negative TrkB-EGFP fusion protein in sparse pyramidal neurons of the adult neocortex and hippocampus, producing a Golgi-staining-like pattern. In adult V1, expression of dominant negative TrkB-EGFP resulted in reduced mushroom spine maintenance and synaptic efficacy, accompanied by an increase in long and thin spines and filopodia. In contrast, mushroom spine maintenance was unaffected in CA1, indicating that TrkB plays fundamentally different roles in structural plasticity in these brain areas.


Asunto(s)
Hipocampo/metabolismo , Receptor trkB/metabolismo , Corteza Visual/metabolismo , Animales , Encéfalo/metabolismo , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Fenómenos Fisiológicos Celulares , ADN/metabolismo , Células Dendríticas/citología , Espinas Dendríticas , Electrofisiología , Genes Dominantes , Aparato de Golgi/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Confocal , Microscopía Fluorescente , Plasticidad Neuronal , Neuronas/metabolismo , Recombinación Genética , Transducción de Señal , Sinapsis/metabolismo , Transmisión Sináptica , Factores de Tiempo
9.
BMC Neurosci ; 5: 4, 2004 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-15005815

RESUMEN

BACKGROUND: Inactivating genes in vivo is an important technique for establishing their function in the adult nervous system. Unfortunately, conventional knockout mice may suffer from several limitations including embryonic or perinatal lethality and the compensatory regulation of other genes. One approach to producing conditional activation or inactivation of genes involves the use of Cre recombinase to remove loxP-flanked segments of DNA. We have studied the effects of delivering Cre to the hippocampus and neocortex of adult mice by injecting replication-deficient adeno-associated virus (AAV) and lentiviral (LV) vectors into discrete regions of the forebrain. RESULTS: Recombinant AAV-Cre, AAV-GFP (green fluorescent protein) and LV-Cre-EGFP (enhanced GFP) were made with the transgene controlled by the cytomegalovirus promoter. Infecting 293T cells in vitro with AAV-Cre and LV-Cre-EGFP resulted in transduction of most cells as shown by GFP fluorescence and Cre immunoreactivity. Injections of submicrolitre quantities of LV-Cre-EGFP and mixtures of AAV-Cre with AAV-GFP into the neocortex and hippocampus of adult Rosa26 reporter mice resulted in strong Cre and GFP expression in the dentate gyrus and moderate to strong labelling in specific regions of the hippocampus and in the neocortex, mainly in neurons. The pattern of expression of Cre and GFP obtained with AAV and LV vectors was very similar. X-gal staining showed that Cre-mediated recombination had occurred in neurons in the same regions of the brain, starting at 3 days post-injection. No obvious toxic effects of Cre expression were detected even after four weeks post-injection. CONCLUSION: AAV and LV vectors are capable of delivering Cre to neurons in discrete regions of the adult mouse brain and producing recombination.


Asunto(s)
Dependovirus/genética , Técnicas de Transferencia de Gen , Integrasas/administración & dosificación , Integrasas/genética , Lentivirus/genética , Neuronas/metabolismo , Proteínas Virales/administración & dosificación , Proteínas Virales/genética , Animales , Línea Celular , Expresión Génica , Genes Reporteros , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes , Hipocampo/metabolismo , Hipocampo/virología , Integrasas/metabolismo , Proteínas Luminiscentes/biosíntesis , Proteínas Luminiscentes/genética , Ratones , Ratones Transgénicos , Neocórtex/metabolismo , Neocórtex/virología , Neuronas/virología , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Recombinación Genética/efectos de los fármacos , Recombinación Genética/genética , Transgenes , Proteínas Virales/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA