Indicadores de cálcio e de voltagem codificados geneticamente na detecção de potenciais de ação e inputs sinápticos em cultura de neurônios hipocampais

Recently, genetically encoded optical indicators have emerged as noninvasive tools of high spatial and temporal resolution utilized to monitor the activity of individual neurons and specific neuronal populations. The increasing number of new optogenetic indicators, together with the absence of comparisons under identical conditions, has generated difficulty in choosing the most appropriate protein, depending on the experimental design. Therefore, the purpose of our study was to compare three recently developed reporter proteins: the calcium indicators GCaMP3 and R-GECO1, and the voltage indicator VSFP butterfly1.2. These probes were expressed in hippocampal neurons in culture, which were subjected to patchclamp recordings and optical imaging. The three groups (each one expressing a protein) exhibited similar values of membrane potential (in mV, GCaMP3: -56 ±8.0, R-GECO1: -57 ±2.5; VSFP: -60 ±3.9, p = 0.86); however, the group of neurons expressing VSFP showed a lower average of input resistance than the other groups (in Mohms, GCaMP3: 161 ±18.3; GECO1-R: 128 ±15.3; VSFP: 94 ±14.0, p = 0.02). Each neuron was submitted to current injections at different frequencies (10 Hz, 5 Hz, 3 Hz, 1.5 Hz, and 0.7 Hz) and their fluorescence responses were recorded in time. In our study, only 26.7% (4/15) of the neurons expressing VSFP showed detectable fluorescence signal in response to action potentials (APs). The average signal-to-noise ratio (SNR) obtained in response to five spikes (at 10 Hz) was small (1.3 ± 0.21), however the rapid kinetics of the VSFP allowed discrimination of APs as individual peaks, with detection of 53% of the evoked APs. Frequencies below 5 Hz and subthreshold signals were undetectable due to high noise. On the other hand, calcium indicators showed the greatest change in fluorescence following the same protocol (five APs at 10 Hz). Among the GCaMP3 expressing neurons, 80% (8/10) exhibited signal, with an average SNR value of 21 ±6.69 (soma), while for the R-GECO1 neurons, 50% (2/4) of the neurons had signal, with a mean SNR value of 52 ±19.7 (soma). For protocols at 10 Hz, 54% of the evoked APs were detected with GCaMP3 and 85% with R-GECO1. APs were detectable in all the analyzed frequencies and fluorescence signals were detected from subthreshold depolarizations as well. Because GCaMP3 is the most likely to yield fluorescence signal and with high SNR, some experiments were performed only with this probe. We demonstrate that GCaMP3 is effective in detecting synaptic inputs (involving Ca2+ influx), with high spatial and temporal resolution. Differences were also observed between the SNR values resulting from evoked APs, compared to spontaneous APs. In recordings of groups of cells, GCaMP3 showed clear discrimination between activated and silent cells, and reveals itself as a potential tool in studies of neuronal synchronization. Thus, our results indicate that the presently available calcium indicators allow detailed studies on neuronal communication, ranging from individual dendritic spines to the investigation of events of synchrony in neuronal networks genetically defined. In contrast, studies employing VSFPs represent a promising technology for monitoring neural activity and, although still to be improved, they may become more appropriate than calcium indicators, since neurons work on a time scale faster than events of calcium may foresee

Evolução morfodinâmica da região de influência estuarina do Rio Curimataú/RN, com ênfase nas alternativas do ambiente deposicional de manguezal e a integração de geodados em Sig

The estuaries are important investigation zones of the actual morphodynamic and of depositional facies of recent geological history. They are constituted in important receptor means of the coastal area sediments, where the evolutionary processes occur quickly. They are also attractive means for the development of anthropic activities, which in a disordered way interfere in the active processes in the sedimentary balance of the coastal areas. Among the human interventions, the alterations of the depositional environment of mangroves in areas of tropical estuary is deserving relevance, whose implications for the environment estuarine and the coastal adjacent, they are still far to be known. Due to the interest of the sedimentologic component in the comprehension of the processes linked to the evolution of the environments estuarine and coastal adjacent, this work, aimed at the understanding of the morphodynamic coastal phenomena that comprise the region of estuarine influence of the River Curimataú / RN. It was also evaluated in the morphodynamic context the implications due to alterations of the depositional environment of mangrove by anthropic activity. The Curimataú Estuary, located in the south portion of the oriental coast of Rio Grande do Norte, in the last decades has been objective of the overwhelming occupation of the shrimp farm in areas of mangroves, which were implanted with perspectives of development in a short to medium period. On the other hand, the estuary and its region of coastal influence lacks enough information to subsidize the planning and reorganization more effective of the surrounding activities. Thus, it was intended with this work to give a contribution target tothe maintainable use of the coastal resources of this region. A series of studies using data of orbital and acoustic remote sensing, as of sediments sampling, were executed in the gutter of the estuary. The obtained results starting from the interpretation of bathymetric maps, echo sounder graphics and of distribution of sediments made possible the location of the estuary based in morpho-sedimentar criteria. The estuarine tidal flat was dissected in environments of intertidal mangroves, supratidal mangroves and apicuns with base in the integration of data of sensor optic and of radar following by the field control. The adjacent coast that is influenced by the Curimataú estuary, was segmented according to their geomorphologic characteristics, where each segment had a point of observation of the beach morphodynamic, during the period from january/2001 to february/2002. Once every month, beaches profiles, collections of sediments in the beach zones, as measurement of hydrodynamic parameters were executed. The results of the observations of the tidal environment showed that the area of estuarine influence of the Curimataú begins to suffer negative sedimentary taxes, where in some beaches, the erosive processes are already observed. The granulometric characteristics of the beach sediments start to tend for the increase of thin sand in the erosive periods. The destruction of the depositional environments of mangroves of the Curimataú estuary, to the construction of shrimp farms, can be providing the diminution of the tidal prism of the estuary, enlarging the effects of the local increasing of the sea level, through the smaller supplying of sediments to the adjacent coast. Besides this, it was verified the possibility of the sanding of the tidal channel in the margins of the destroyed areas of mangroves, where very high taxes of sedimentation of thin materials were estimated in case of these areas were preserve

A erosão costeira e sua influência sobre a atividade petrolífera: alternativas sustentáveis na mitigação de impactos ambientais

The study area consist in high sensitivity environments located on the northern coast of Rio Grande do Norte, Northeast Brazil. The barrier island are the main geomorphological features on the coastal landscape, being naturally instable and surrounded by industrial activities like oil fields, salt industry, shrimp farms and urban areas sometimes installed parallel to the coast, combined with coast engineering interventions. High energy hydrodynamic process are responsible for the morphological instability of the coast. The study was based on remote sensing data obtained between 1954 and 2007 which consist in orbital images from Landsat, CBERS and Ikonos satellites and aerial photos. With all data integrated on GIS environment it was possible to update thematic maps of geology, geomorphology, vegetation, soil and landuse and development of multitemporal maps pointing areas with erosion and depositions of sediments, defining the critical erosion process on this region. The bigger morphological changes are related to changes on wind patterns during the year, terrestrial and sea breezes during the day, with spits and barrier island migration, opening and closing of channels like the one parallel to the coast on the area of Serra and Macau oil fields. These factors combined with the significant reduction on sediment budgets due to the loss of natural spaces to sediment reworking contribute to the low resilience which tends to be growing on the area of Serra and Macau oil fields. In front of such scenery a detailed monitoring was done in order to find technological possibilities for coastal restoration. A pilot area was defined to start the project of mangrove restore together with beach nourishment in order to minimize the effect of the erosion caused by the channel parallel to the coast, contributing to stabilize the northeast channel as the main one. It s expected that such methodology will aid the coastal environments restoration and the balance between industrial activities and coastal erosion

Análise morfométrica e ultra-estrutural do nervo óptico de ratos induzidos a ingestão de álcool

OBJETIVO: Analisar os efeitos da ingestão crônica de álcool sobre o nervo óptico em um modelo murino adulto. MÉTODOS: Doze ratos machos da raça Wistar, de 30 dias de idade, foram divididos por sorteio em 2 grupos experimentais: tratado (TG), com 8 animais, alimentados com ração-padrão para roedores de laboratório e uma mistura de água de torneira e etanol ad libitum; controle (CG), com 4 animais, alimentados com a mesma ração e água de torneira pura ad libitum. Após 40 semanas todos os ratos foram sacrificados, sendo os nervos ópticos de ambos os olhos preparados para microscopia óptica e eletrônica. A área de secção transversal de cada nervo a aumento de 500´, assim como número de fibras axonais dentro de 5 campos aleatoriamente selecionados a aumento de 2000´ foram medidos com auxílio de digitalizador de imagens acoplado ao microscópio óptico. Foram realizadas fotomicrografias de 10 campos aleatoriamente selecionados de cada nervo (5 centrais e 5 periféricos) a aumento de 4200´ em microscópio eletrônico de transmissão. RESULTADOS: A análise morfométrica não mostrou diferenças estatisticamente significativas entre os 2 grupos estudados. em contraste com o CG, o exame ultra-estrutural dos nervos ópticos do TG mostrou um intenso desarranjo das bainhas de mielina, que se tornaram espessadas, com separação de suas lamelas, apresentando, por vezes, degenerações interlamelares elétron-densas, além da presença de muitas organelas degeneradas. CONCLUSÃO: Os achados desse estudo mostram alterações ultra-estruturais no nervo óptico de ratos adultos após ingestão crônica de álcool, sem modificações morfométricas significativas.

Inclusão social do deficiente visual: experiência e resultados de Assis

OBJETIVOS: Traçar o perfil do usuário da Sala de Recursos para Deficiente Visual na cidade de Assis - SP, avaliar a porcentagem de deficientes visuais empregados e comparar o emprego com idade, gênero, tipo de deficiência e doença causadora da deficiência, entre dois períodos: de 1984 a 1996 e de 1997 a 2009. MÉTODOS: Foi realizado estudo retrospectivo dos prontuários médicos e da ficha escolar dos portadores de deficiência visual que frequentaram a Sala de Recursos para Deficiente Visual, no período de 1984 a 2009 na cidade de Assis - SP, divididos em dois períodos de 1984 a 1996 (G1) e de 1997 a 2009 (G2). Analisaram-se dados demográficos, a doença que provocou a baixa visão, o tipo de deficiência visual (cegueira ou baixa visão), escolaridade, recursos ópticos, frequência à Sala de Recursos para Deficiente Visual e taxa de emprego. Foi feita associação da taxa de emprego com: idade, gênero, raça/cor, tipo de deficiência e doença nos dois diferentes períodos. RESULTADOS: Foram encontrados 149 deficientes visuais sendo: 61,07% homens, 38,9% mulheres, 82,5% brancos e 17,4% não brancos e a mediana da idade foi de 18 anos. Dos 149 deficientes visuais, 63,75% eram portadores de baixa visão e 36,24% portadores de cegueira. As principais doenças que levaram à deficiência visual dos 149 pacientes foram em ordem decrescente: retinocoroidite por toxoplasmose (17,40%), atrofia óptica congênita (12,10%), alta hipermetropia (8,72%), retinose pigmentar e alta miopia (6,71% cada uma) e glaucoma congênito e catarata congênita (6,04% cada uma). A frequência à Sala de Recursos para Deficiente Visual foi boa em mais de 50% dos pacientes. Estavam trabalhando regularmente 44,7% e 12,3% dos maiores de 14 anos respectivamente nos períodos de 1984 - 1996 e de 1997 - 2009. Não houve diferença entre os dois períodos quanto às características demográficas, tipo de deficiência e doenças, havendo correlação da taxa de emprego apenas com a idade (pacientes com média de idade maior apresentavam maior porcentagem de emprego). CONCLUSÕES: Os deficientes visuais eram na maioria homens, brancos, portadores de baixa visão e portadores de toxoplasmose ocular. Quase metade (44,7%) dos pacientes com mais de 14 anos encontrava-se trabalhando regularmente até 1996 sugerindo que esforços conjuntos de oftalmologistas e educadores auxilia na inclusão social destes pacientes. A taxa de emprego caiu no período de 1997 a 2009 e a de aposentados aumentou.

Síntese e caracterização de esferas monodispersas de látex e sua utilização no preparo de filmes de cristais coloidais tridimensionais

Esferas de poliestireno foram obtidas através da polimerização do estireno sem a presença de emulsificante. A suspensão foi caracterizada por Espalhamento de Luz e Microscopia Eletrônica de Transmissão apresentando monodispersidade com tamanho em torno de 463 nm. O método de deposição vertical foi utilizado para preparar filmes em substratos de vidro a partir da suspensão de esferas. Filmes de opalas de alta qualidade foram obtidos e caracterizados por Espectroscopia de Reflectância, Microscopia Eletrônica de Varredura e Microscopia Óptica.

Sintering of tin oxide and its applications in electronics and processing of high purity optical glasses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Visible emission from Er-doped SnO2 thin films deposited by sol-gel

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Imagens multitemporais ALOS/PALSAR baseada em orientação a objeto na discriminação da cobertura da terra

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influência da injunção da base na fototriangulação de imagens obtidas com uma unidade de mapeamento móvel

Pós-graduação em Ciências Cartográficas - FCT

Análise teórica de uma nova técnica de processamento de sinais interferométricos baseada na modulação triangular da fase óptica

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sensor eletro-óptico de tensões elevadas e sua viabilidade para implementação de TP óptico

Pós-graduação em Engenharia Elétrica - FEIS

Identificação do teor de nitrogênio foliar em cana-de-açúcar no estágio inicial de crescimento utilizando infravermelho próximo e sistemas inteligentes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Sensor em fibra óptica aplicado à caracterização de atuadores piezoelétricos flextensionais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos de doses subletais do imidaclopride no cérebro, ventrículo e túbulo de Malpighi de Apis melifera africanizada

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)