RESUMO
Lysosomes are cell organelles that degrade macromolecules to recycle their components. If lysosomal degradative function is impaired, e.g., due to mutations in lysosomal enzymes or membrane proteins, lysosomal storage diseases (LSDs) can develop. LSDs manifest often with neurodegenerative symptoms, typically starting in early childhood, and going along with a strongly reduced life expectancy and quality of life. We show here that small molecule activation of the Ca2+ -permeable endolysosomal two-pore channel 2 (TPC2) results in an amelioration of cellular phenotypes associated with LSDs such as cholesterol or lipofuscin accumulation, or the formation of abnormal vacuoles seen by electron microscopy. Rescue effects by TPC2 activation, which promotes lysosomal exocytosis and autophagy, were assessed in mucolipidosis type IV (MLIV), Niemann-Pick type C1, and Batten disease patient fibroblasts, and in neurons derived from newly generated isogenic human iPSC models for MLIV and Batten disease. For in vivo proof of concept, we tested TPC2 activation in the MLIV mouse model. In sum, our data suggest that TPC2 is a promising target for the treatment of different types of LSDs, both in vitro and in-vivo.
Assuntos
Doenças por Armazenamento dos Lisossomos , Mucolipidoses , Lipofuscinoses Ceroides Neuronais , Animais , Pré-Escolar , Humanos , Lisossomos/metabolismo , Camundongos , Mucolipidoses/genética , Mucolipidoses/metabolismo , Lipofuscinoses Ceroides Neuronais/metabolismo , Qualidade de VidaRESUMO
Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are dually gated channels that are operated by voltage and by neurotransmitters via the cAMP system. cAMP-dependent HCN regulation has been proposed to play a key role in regulating circuit behavior in the thalamus. By analyzing a knockin mouse model (HCN2EA), in which binding of cAMP to HCN2 was abolished by 2 amino acid exchanges (R591E, T592A), we found that cAMP gating of HCN2 is essential for regulating the transition between the burst and tonic modes of firing in thalamic dorsal-lateral geniculate (dLGN) and ventrobasal (VB) nuclei. HCN2EA mice display impaired visual learning, generalized seizures of thalamic origin, and altered NREM sleep properties. VB-specific deletion of HCN2, but not of HCN4, also induced these generalized seizures of the absence type, corroborating a key role of HCN2 in this particular nucleus for controlling consciousness. Together, our data define distinct pathological phenotypes resulting from the loss of cAMP-mediated gating of a neuronal HCN channel.
Assuntos
AMP Cíclico/metabolismo , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/genética , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/metabolismo , Convulsões/metabolismo , Animais , Comportamento Animal , Epilepsia/metabolismo , Células HEK293 , Humanos , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Moleculares , Neurônios/metabolismo , Canais de Potássio , Tálamo/metabolismo , TranscriptomaRESUMO
Hyperpolarization-activated cyclic nucleotide-gated channels (HCNs) in the nervous system are implicated in a variety of neuronal functions including learning and memory, regulation of vigilance states and pain. Dysfunctions or genetic loss of these channels have been shown to cause human diseases such as epilepsy, depression, schizophrenia, and Parkinson's disease. The physiological functions of HCN1 and HCN2 channels in the nervous system have been analyzed using genetic knockout mouse models. By contrast, there are no such genetic studies for HCN3 channels so far. Here, we use a HCN3-deficient (HCN3-/-) mouse line, which has been previously generated in our group to examine the expression and function of this channel in the CNS. Specifically, we investigate the role of HCN3 channels for the regulation of circadian rhythm and for the determination of behavior. Contrary to previous suggestions we find that HCN3-/- mice show normal visual, photic, and non-photic circadian function. In addition, HCN3-/- mice are impaired in processing contextual information, which is characterized by attenuated long-term extinction of contextual fear and increased fear to a neutral context upon repeated exposure.
RESUMO
Numerous signal pathways are epigenetically controlled during brain development and ageing. Thereby, both 5-methylcytosine (5mC) and the newly described 5-hydroxymethylcytosine (5hmC) are highly exhibited in the brain. As there is an uneven distribution of 5hmC in the brain depending on age and region, there is the need to investigate the underlying mechanisms being responsible for 5hmC generation and decline. The aim of this study was to quantify expression levels of genes that are associated with DNA methylation/demethylation in different brain regions and at different ages. Therefore, we investigated frontal cortex and cerebellum of 40 mice (strain C57BL/6), each eight mice sacrificed at day 0, 7, 15, 30 and 120 after birth. We performed expression profiling of methylation/demethylation genes depending on age and brain region. Interestingly, we see significant expression differences of genes being responsible for methylation/demethylation with a significant reduction of expression levels during ageing. Validating selected expression data on protein level using immunohistochemistry verified the expression data. In conclusion, our findings demonstrate that the regulation of methylation/demethylation pathways is highly controlled depending on brain region and age. Thus our data will help to better understand the complexity and plasticity of the brain epigenome.
Assuntos
Envelhecimento/fisiologia , Encéfalo/crescimento & desenvolvimento , Metilação de DNA/fisiologia , 5-Metilcitosina/metabolismo , Animais , Encéfalo/metabolismo , Citosina/análogos & derivados , Citosina/metabolismo , Epigênese Genética/fisiologia , Feminino , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase , TranscriptomaRESUMO
Continuous time-series estimates of net ecosystem carbon exchange (NEE) are routinely made using eddy covariance techniques. Identifying and compensating for errors in the NEE time series can be automated using a signal processing filter like the ensemble Kalman filter (EnKF). The EnKF compares each measurement in the time series to a model prediction and updates the NEE estimate by weighting the measurement and model prediction relative to a specified measurement error estimate and an estimate of the model-prediction error that is continuously updated based on model predictions of earlier measurements in the time series. Because of the covariance among model variables, the EnKF can also update estimates of variables for which there is no direct measurement. The resulting estimates evolve through time, enabling the EnKF to be used to estimate dynamic variables like changes in leaf phenology. The evolving estimates can also serve as a means to test the embedded model and reconcile persistent deviations between observations and model predictions. We embedded a simple arctic NEE model into the EnKF and filtered data from an eddy covariance tower located in tussock tundra on the northern foothills of the Brooks Range in northern Alaska, USA. The model predicts NEE based only on leaf area, irradiance, and temperature and has been well corroborated for all the major vegetation types in the Low Arctic using chamber-based data. This is the first application of the model to eddy covariance data. We modified the EnKF by adding an adaptive noise estimator that provides a feedback between persistent model data deviations and the noise added to the ensemble of Monte Carlo simulations in the EnKF. We also ran the EnKF with both a specified leaf-area trajectory and with the EnKF sequentially recalibrating leaf-area estimates to compensate for persistent model-data deviations. When used together, adaptive noise estimation and sequential recalibration substantially improved filter performance, but it did not improve performance when used individually. The EnKF estimates of leaf area followed the expected springtime canopy phenology. However, there were also diel fluctuations in the leaf-area estimates; these are a clear indication of a model deficiency possibly related to vapor pressure effects on canopy conductance.
Assuntos
Carbono/química , Modelos Teóricos , Regiões ÁrticasRESUMO
Feedbacks between water use, biomass and infiltration capacity in semiarid ecosystems have been shown to lead to the spontaneous formation of vegetation patterns in a simple model. The formation of patterns permits the maintenance of larger overall biomass at low rainfall rates compared with homogeneous vegetation. This results in a bias of models run at larger scales neglecting subgrid-scale variability. In the present study, we investigate the question whether subgrid-scale heterogeneity can be parameterized as the outcome of optimal partitioning between bare soil and vegetated area. We find that a two-box model reproduces the time-averaged biomass of the patterns emerging in a 100 x 100 grid model if the vegetated fraction is optimized for maximum entropy production (MEP). This suggests that the proposed optimality-based representation of subgrid-scale heterogeneity may be generally applicable to different systems and at different scales. The implications for our understanding of self-organized behaviour and its modelling are discussed.
Assuntos
Clima Desértico , Ecossistema , Entropia , Modelos Teóricos , Desenvolvimento Vegetal , Chuva , Retroalimentação , TermodinâmicaRESUMO
This study explores the relationship between the normalized difference vegetation index (NDVI) and aboveground plant biomass for tussock tundra vegetation and compares it to a previously established NDVI-biomass relationship for wet sedge tundra vegetation. In addition, we explore inter-annual variation in NDVI in both these contrasting vegetation communities. All measurements were taken across long-term experimental treatments in wet sedge and tussock tundra communities at the Toolik Lake Long Term Ecological Research (LTER) site, in northern Alaska. Over 15 years (for wet sedge tundra) and 14 years (for tussock tundra), N and P were applied in factorial experiments (N, P and N+P), air temperature was increased using greenhouses with and without N+P fertilizer, and light intensity was reduced by 50% using shade cloth. during the peak growing seasons of 2001, 2002, and 2003, NDVI measurements were made in both the wet sedge and tussock tundra experimental treatment plots, creating a 3-year time series of inter-annual variation in NDVI. We found that: (1) across all tussock experimental tundra treatments, NDVI is correlated with aboveground plant biomass (r2 = 0.59); (2) NDVI-biomass relationships for tussock and wet sedge tundra communities are community specific, and; (3) NDVI values for tussock tundra communities are typically, but not always, greater than for wet sedge tundra communities across all experimental treatments. We suggest that differences between the response of wet sedge and tussock tundra communities in the same experimental treatments result from the contrasting degree of heterogeneity in species and functional types that characterize each of these Arctic tundra vegetation communities.
Assuntos
Ecossistema , Meio Ambiente , Fenômenos Fisiológicos Vegetais , Temperatura , Alaska , Análise de Variância , Biomassa , Estudos Longitudinais , Nitrogênio , FósforoRESUMO
We show that the spatial-temporal variability of human West Nile (WN) cases and the transmission of West Nile virus (WNV) to sentinel chickens are associated with the spatial-temporal variability of drought and wetting in southern Florida. Land surface wetness conditions at 52 sites in 31 counties in southern Florida for 2001-2003 were simulated and compared with the occurrence of human WN cases and the transmission of WNV to sentinel chickens within these counties. Both WNV transmission to sentinel chickens and the occurrence of human WN cases were associated with drought 2-6 mo prior and land surface wetting 0.5-1.5 mo prior. These dynamics are similar to the amplification and transmission patterns found in southern Florida for the closely related St. Louis encephalitis virus. Drought brings avian hosts and vector mosquitoes into close contact and facilitates the epizootic cycling and amplification of the arboviruses within these populations. Southern Florida has not recorded a severe, widespread drought since the introduction of WNV into the state in 2001. Our results indicate that widespread drought in the spring followed by wetting during summer greatly increase the probability of a WNV epidemic in southern Florida.
Assuntos
Desastres , Febre do Nilo Ocidental/epidemiologia , Febre do Nilo Ocidental/transmissão , Vírus do Nilo Ocidental , Animais , Galinhas , Florida , Humanos , Modelos Logísticos , Doenças das Aves Domésticas/transmissão , Doenças das Aves Domésticas/virologia , Febre do Nilo Ocidental/veterináriaRESUMO
Using a dynamic hydrology model, we simulated land surface wetness conditions at 42 sites in 28 counties in southcentral Florida from 1990 to 1998 and compared these simulations with the incidence of human cases of St. Louis encephalitis (SLE) within these counties. Within counties, drought four months prior and wetting one-half month prior were significantly associated with human cases of SLE. Simulated land surface wetness conditions resolved transmission loci in both space and time, and May drought was significantly associated with the subsequent occurrence of human SLE cases. These findings are consistent with previous results associating simulated land surface wetness conditions with the transmission of SLE virus as measured in sentinel chickens, and support our working hypothesis that springtime drought facilitates SLE virus amplification in mosquito and wild bird populations.
Assuntos
Desastres/estatística & dados numéricos , Surtos de Doenças/estatística & dados numéricos , Encefalite de St. Louis/epidemiologia , Demografia , Florida/epidemiologia , Humanos , Modelos Teóricos , Chuva , Estudos Retrospectivos , Fatores de Tempo , Tempo (Meteorologia)RESUMO
Disease transmission forecasts can help minimize human and domestic animal health risks by indicating where disease control and prevention efforts should be focused. For disease systems in which weather-related variables affect pathogen proliferation, dispersal, or transmission, the potential for disease forecasting exists. We present a seasonal forecast of St. Louis encephalitis virus transmission in Indian River County, Florida. We derive an empiric relationship between modeled land surface wetness and levels of SLEV transmission in humans. We then use these data to forecast SLEV transmission with a seasonal lead. Forecast skill is demonstrated, and a real-time seasonal forecast of epidemic SLEV transmission is presented. This study demonstrates how weather and climate forecast skill-verification analyses may be applied to test the predictability of an empiric disease forecast model.
Assuntos
Vírus da Encefalite de St. Louis/isolamento & purificação , Encefalite de St. Louis/transmissão , Previsões , Modelos Biológicos , Estações do Ano , Animais , Galinhas/virologia , Clima , Culex/virologia , Encefalite de St. Louis/epidemiologia , Encefalite de St. Louis/virologia , Feminino , Florida/epidemiologia , Humanos , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/transmissão , Doenças das Aves Domésticas/virologia , Vigilância de Evento Sentinela , Tempo (Meteorologia)RESUMO
We analyzed the prevalence of hemagglutination inhibition (HI) antibodies to St. Louis encephalitis (SLE) virus in wild birds during the 1990 SLE epidemic in Indian River County. The initial presence of SLE HI antibody was associated significantly with modeled drought 15 wk prior, wetting conditions 1 wk prior, and the emergence of the Florida SLE virus vector, Culex nigripalpus, 5 wk prior. Our findings indicated that three factors conspired to create the 1990 epidemic: (1) a large population of susceptible wild birds; (2) severe springtime drought, which facilitated amplification of the SLE virus among the Cx. nigripalpus and a portion of the wild bird population; and (3) continued rainfall and wetting of the land surface in the summer and early fall, which sustained a large, host-seeking Cx. nigripalpus population. The continued biting and reproductive activity of Cx. nigripalpus maintained epizootic transmission throughout the summer and early fall in Indian River County. The high level of SLE virus amplification resulted in spillover transmission to humans. We hypothesize that without the continued reproductive activity of the vector mosquito, brought about by excessive summer and fall wetness, the unprecedented SLE virus amplification and consequent transmission to humans would not have been realized in 1990.
Assuntos
Aves/virologia , Culex/crescimento & desenvolvimento , Culex/virologia , Vírus da Encefalite de St. Louis/isolamento & purificação , Encefalite de St. Louis/epidemiologia , Animais , Animais Selvagens/virologia , Anticorpos Antivirais/sangue , Clima , Desastres , Vírus da Encefalite de St. Louis/patogenicidade , Feminino , Florida/epidemiologia , Densidade Demográfica , Chuva , Estações do AnoRESUMO
This study explores the relationship between the normalized difference vegetation index (NDVI), aboveground plant biomass, and ecosystem C fluxes including gross ecosystem production (GEP), ecosystem respiration (ER) and net ecosystem production. We measured NDVI across long-term experimental treatments in wet sedge tundra at the Toolik Lake LTER site, in northern Alaska. Over 13 years, N and P were applied in factorial experiments (N, P and N + P), air temperature was increased using greenhouses with and without N + P fertilizer, and light intensity (photosynthetically active photon flux density) was reduced by 50% using shade cloth. Within each treatment plot, NDVI, aboveground biomass and whole-system CO(2) flux measurements were made at the same sampling points during the peak-growing season of 2001. We found that across all treatments, NDVI is correlated with aboveground biomass ( r(2)=0.84), GEP ( r(2)=0.75) and ER ( r(2)=0.71), providing a basis for linking remotely sensed NDVI to aboveground biomass and ecosystem carbon flux.
Assuntos
Dióxido de Carbono/análise , Dióxido de Carbono/metabolismo , Fotossíntese , Biomassa , Ecossistema , Monitoramento Ambiental , Gases , Nitrogênio/metabolismo , Fósforo/metabolismo , Fenômenos Fisiológicos Vegetais , AstronaveRESUMO
We used a dynamic hydrology model to simulate water table depth (WTD) and quantify the relationship between Saint Louis encephalitis virus (SLEV) transmission and hydrologic conditions in Indian River County, Florida, from 1986 through 1991, a period with an SLEV epidemic. Virus transmission followed periods of modeled drought (specifically low WTDs 12 to 17 weeks before virus transmission, followed by a rising of the water table 1 to 2 weeks before virus transmission). Further evidence from collections of Culex nigripalpus (the major mosquito vector of SLEV in Florida) suggests that during extended spring droughts vector mosquitoes and nestling, juvenile, and adult wild birds congregate in selected refuges, facilitating epizootic amplification of SLEV. When the drought ends and habitat availability increases, the SLEV-infected Cx. nigripalpus and wild birds disperse, initiating an SLEV transmission cycle. These findings demonstrate a mechanism by which drought facilitates the amplification of SLEV and its subsequent transmission to humans.
Assuntos
Galinhas/virologia , Desastres , Vírus da Encefalite de St. Louis/isolamento & purificação , Encefalite de St. Louis/epidemiologia , Modelos Teóricos , Microbiologia da Água , Animais , Anticorpos Antivirais/sangue , Culex/virologia , Encefalite de St. Louis/transmissão , Encefalite de St. Louis/virologia , Doenças Endêmicas , Feminino , Florida/epidemiologia , Testes de Inibição da Hemaglutinação , Modelos Logísticos , MasculinoRESUMO
We modeled surface wetness at high resolution, using a dynamic hydrology model, to predict flood and swamp water mosquito abundances. Historical meteorologic data, as well as topographic, soil, and vegetation data, were used to model surface wetness and identify potential fresh and swamp water breeding habitats in two northern New Jersey watersheds. Surface wetness was positively associated with the subsequent abundance of the dominant floodwater mosquito species, Aedes vexans, and the swamp water species, Anopheles walkeri. The subsequent abundance of Culex pipiens, a species that breeds in polluted, eutrophic waters, was negatively correlated with local modeled surface wetness. These associations permit real-time monitoring and forecasting of these floodwater and nonfloodwater species at high spatial and temporal resolution. These predictions will enable public health agencies to institute control measures before the mosquitoes emerge as adults, when their role as transmitters of disease comes into play.