Name: Padronização de um novo software semiautomático para medição de crescimento de neuritos (Vídeo) | JoVE Journal
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Description: Aprenda a medir crescimento de neuritos com software semiautomático em modelos in vitro de dor neuropática. Este método padronizado analisa dinâmica de importin alpha-3 em camundongos após lesão espinhal.

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<meta charset="utf8"></head><body>Técnicas avançadas de imagem ao vivo para medição precisa do crescimento de neuritos

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<li>Ackerman, H. D., Gerhard, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bile+acids+induce+neurite+outgrowth+in+NSC-34+cells+via+TGR5+and+a+distinct+transcriptional+profile%5BTitle%5D)+AND+%22Pharmaceuticals%22%5BJournal%5D)">Bile acids induce neurite outgrowth in NSC-34 cells via TGR5 and a distinct transcriptional profile.</a> Pharmaceuticals. 16, 174(2023).</li>
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</ol>

<table><tbody><tr><td>Colagenase A</td><td>Merck / Roche</td><td>10103586001</td><td></td></tr><tr><td>Dispase II (protease neutra, grau II)</td> <td>Merck / Roche</td><td>4942078001</td><td></td></tr><tr><td>meio de águia modificado de Dulbecco</td><td>Merck / Sigma</td><td>D5796</td><td></td></tr><tr><td>Soro fetal de bovin&nbsp;</td><td>Merck / Sigma</td><td>F7524</td><td></td></tr><tr><td>Mistura de Nutrientes F-12 da Ham (1X)</td><td>ThermoFisher Scientific</td><td>21765029</td><td></td></tr><tr><td>F12 da Ham's com L-Glutamina</td><td>Euroclone</td><td>ECM0135L</td><td></td></tr><tr><td>Hanks' Solução Salar Balanceada</td><td>Euroclone</td><td>ECM0507L</td><td></td></tr><tr><td>HBSS (10X), sem cálcio, sem magnésio, sem vermelho de fenol</td><td>ThermoFisher Scientific</td><td>14185045</td><td></td></tr><tr><td>HyClone caracterizou soro fetal bovino (EUA)</td><td>Cytiva</td><td>SH30071.03</td><td></td></tr><tr><td>Incucyte, Software de Análise Neurotrack&nbsp;</td><td>Sartorius</td><td>9600-0010</td><td></td></tr><tr><td>L-15 Medium (Leibovitz)</td><td>Millipore/Sigma</td><td>L5520</td><td></td></tr><tr><td>Laminin Protein Rat, Natural</td><td>ThermoFisher Scientific</td><td>23017015</td><td></td></tr><tr><td>L-cisteína</td><td>Merck / Sigma</td><td>C7352</td><td></td></tr><tr><td>Leibovitz's L-15 medium s/o L-glutamine</td><td>Euroclone</td><td>ECB0020L</td><td></td></tr><tr><td>mouse NGF 2.5S (>95%)</td><td>Alomone Labs</td><td>N-100</td><td></td></tr><tr><td>Meio Neurobasal [-] Glutamina</td><td>ThermoFisher Scientific</td><td>21103049</td><td></td></tr><tr><td>NSC-34</td><td>CELLutions Biosystems Inc (Ontário, Canadá)</td><td>CLU140</td><td></td></tr><tr><td>Papaína do látex de mamão</td><td>Sigma</td><td>P4762</td><td></td></tr><tr><td>Penicilina-Estreptomicina (5.000 U/mL)</td><td>ThermoFisher Scientific</td><td>15070063</td><td></td></tr><tr><td>Percoll ( Densidade 1,130 g / mL)</td> <td>Cytiva</td><td>17089101</td><td></td></tr><tr><td>Solução de Poli-D-Lisina (1mg / mL)</td> <td>EMD Millipore / Merck</td><td>A-003-E</td><td></td></tr><tr><td>Solução de Poli-L-Lisina (0-01%)</td><td>Sigma</td><td>P4832</td><td></td></tr><tr><td>Humano Recombinante NT-3</td><td>PeproTech</td><td>450-03</td><td></td></tr><tr><td>Ácido Retinóico</td><td>Merck / Sigma</td><td>R2625</td><td></td></tr><tr><td>Solução de tripsina-EDTA</td><td>Sigma</td><td>T3924</td><td></td></tr><tr><td>&beta;-Tubulina III (Tuj1) anticorpo</td><td>Merck / Sigma</td><td>T8660</td><td></td></tr></tbody></table>

padronização de um novo software semiautomático para medição de crescimento de neuritos 

Standardization of a Novel Semi-Automatic Software for Neurite Outgrowth Measurement

Padronização de um novo software semiautomático para medição de crescimento de neuritos 

Effective live-imaging techniques are crucial to assess neuronal morphology in order to measure neurite outgrowth in real time. The proper measurement of ...

standardization-novel-semi-automatic-software-for-neurite-outgrowth

Research

JoVE Journal

Neuroscience

Neurociência

A abundância de receptores de neurotransmissores nas sinapses cluster influencia fortemente a força sináptica. Este método quantifica receptores de neurotransmissores fluorescente-etiquetados em três dimensões com um único sinapse resolução...

Quantificação automatizada de fluorescência em Synaptic<em&gt; C. elegans</em

Este vídeo demonstra o ensaio de neuroplasticidade in vitro para estudar as alterações morfológicas nos gânglios da raiz dorsal e nos neurônios do plexo mioentérico expostos a extratos de tecido pancreático humano saudável e canceroso. Este ensaio nos permite estudar o crescimento de neuritos, o padrão de ramificação e o tamanho neuronal.

Ensaio de neuroplasticidade pancreática in vitro: estudando alterações nos neurônios do trato gastrointestinal expostos a extratos de tecido pancreático humano saudável e canceroso

Plasticidade neuronal é uma característica cada vez mais reconhecida, mas não suficientemente entendido do gastrointestinal (GI). Aqui, no exemplo de desordens pancreáticas humanas, apresentamos um ensaio neuroplasticidade in vitro para o estudo da plasticidade neuronal no tracto gastrointestinal, a nível tanto morfológica e funcional.

Simulando pâncreas Neuroplasticidade:<em&gt; In Vitro</em&gt; Dual-neurônio Plasticidade Assay

Com a intenção de caracterizar mudanças em miRNAs em células-tronco neurais humanas diferenciadas (hNSCs), descrevemos a diferenciação de hNSC em um sistema tridimensional, a avaliação de mudanças na expressão de microRNAs por matriz de PCR de miRNA e análise computacional para predição de alvos de miRNAs e sua validação por ensaio de luciferase dupla.

Diferenciação de uma linha de células-tronco neurais em Três Culturas dimensionais, os genes Análise de microRNA e putativo alvo

Neste relatório, nós descrevemos um protocolo simples para estudar o conseqüência do neurite em neurônios corticais do rato embrionário cotransfecting com EGFP e a proteína do interesse.

Um ensaio com base em proteínas de fluorescência verde reforçada para estudar o crescimento do neurite em neurônios primários

O protocolo apresentado descreve um método para um ensaio de crescimento de neurite e avaliação neurotoxicidade de compostos de moléculas pequenas.

Um ensaio de crescimento de neurita e avaliação de neurotoxicidade com neurônios derivados de células progenitoras neugenitoras humanas

Aqui, apresentamos um protocolo detalhado para estudar o acúmulo neuronal de α-sinucleína em neurônios primários de dopamina de camundongos. Os agregados fosforilados de α-sinucleína em neurônios são induzidos com fibrilas pré-formadas de α-sinucleína. Imagens automatizadas de células imunofluorescentamente rotuladas e análise de imagem imparcial tornam este protocolo robusto adequado para a triagem de rendimento médio a alto de drogas que inibem o acúmulo de α-sinucleína.

Estudando o acúmulo de α-sinucleína induzida por fibril pré-formado em neurônios de dopamina de cérebro médio do camundongo embrionário primário

Mostramos a automação das culturas de células-tronco pluripotentes induzidas pelo homem (hiPSC) e diferenciações neuronais compatíveis com imagens e análises automatizadas.

Produção automatizada de neurônios cortical e dopaminérgico derivados de células-tronco pluripotentes induzidos por humanos com monitoramento integrado de células vivas

Aqui apresentamos um método automatizado para determinação semi-quantitativa do número de neurônio dopaminérgico na substantia nigra pars compacta.

Determinação semi-quantitativa da densidade do neurônio dopaminérgico na Substantia Nigra dos Modelos de Roedores usando análise automatizada de imagem

Apresentado é uma ferramenta computacional que permite a medição automática simples e direta das orientações de ramos dendríticas neuronais a partir de imagens de fluorescência 2D.

Identificação Automática de Ramos Dendráticos e sua Orientação

As espinhas dendríticas são compartimentos pós-sinápticos da maioria das sinapses excitatórias. Alterações na morfologia da coluna dendrítica ocorrem durante o neurodesenvolvimento, envelhecimento, aprendizado e muitos distúrbios neurológicos e psiquiátricos, ressaltando a importância de uma análise confiável da coluna dendrítica. Este protocolo descreve a quantificação da morfologia da coluna dendrítica de forma precisa e reprodutível usando um software automático de reconstrução...

Quantificação da coluna dendrítica usando um software automático de reconstrução tridimensional de neurônios

Aprenda a medir crescimento de neuritos com software semiautomático em modelos in vitro de dor neuropática. Este método padronizado analisa dinâmica de importin alpha-3 em camundongos após lesão espinhal.

Padronização de um novo software semiautomático para medição de crescimento de neuritos

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