Name: measuring near plasma membrane and global intracellular calcium dynamics in astrocytes
Uploaded: 2009-04-26T23:27:00
Description: Watch this Scientific Journal Video about Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes at JoVE.com

Este trabalho foi financiado pela Fundação Memorial Uehara do Japão (a ES), bem como a Fundação Whitehall, do Instituto Nacional de Distúrbios Neurológicos e Derrame e um Stein-Oppenheimer Award Endowment (a BSK).

Procedimentos experimentais O procedimento experimental consiste de duas partes principais que são descritas em uma maneira passo sábio abaixo. Parte 1: PREPARAÇÃO cultura de astrócitos hipocampais Brevemente, misturado astrócitos do hipocampo neurônio-culturas foram preparadas usando um protocolo bem estabelecido 1,2,3. Nós otimizamos o procedimento para produzir saudável astrócitos em cultura. Todos os procedimentos...

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É bem estabelecido que os astrócitos mostrar elevações de cálcio intracelular. Estas ocorrem de forma espontânea, pode ser desencadeada pela atividade neuronal ou por aplicação de agonistas para ativar receptores na superfície de astrócitos 11. Uma questão importante e controversa é se astrócitos elevações de cálcio intracelular pode desencadear a liberação de moléculas de sinalização que ativam os receptores nos neurônios 11, 12. Este é controverso, porque tem havido evidências a favor e contra...

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<td>Invitrogen</td>
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<td>Immersion Oil TYPE DF</td>
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measuring near plasma membrane and global intracellular calcium dynamics in astrocytes

O cérebro contém células da glia. Astrócitos, um tipo de célula glial, tem sido conhecido por fornecer um papel passivo de apoio aos neurônios. No entanto, evidências crescentes sugerem que os astrócitos também podem participar ativamente na função cerebral por meio de interações funcionais com os neurônios. No entanto, muitos aspectos fundamentais da biologia astrócitos permanecem controversas, pouco clara e / ou experimentalmente inexplorado. Uma questão importante é a dinâmica dos transientes de cálcio intracelular em astrócitos. Isso é relevante porque o cálcio é bem estabelecido como um importante mensageiro segundo e porque tem sido proposto que a elevação de cálcio astrócitos pode desencadear a liberação de transmissores de astrócitos. No entanto, não houve qualquer descrição detalhada ou satisfatória de cálcio perto da membrana plasmática de sinalização em astrócitos. Reflexão interna total de fluorescência microscopia (TIRF) é uma ferramenta poderosa para analisar fisiologicamente relevantes eventos de sinalização dentro de cerca de 100 nm da membrana plasmática de células vivas. Aqui, usamos a microscopia TIRF e descrever como monitor perto de membrana plasmática e global a dinâmica do cálcio intracelular quase simultaneamente. O aperfeiçoamento e aplicação sistemática desta abordagem tem o potencial de informar sobre os detalhes precisos de sinalização de cálcio astrócitos. Um entendimento detalhado da dinâmica de cálcio astrócitos pode fornecer uma base para entender se, como, quando e por que astrócitos e neurônios sofrem de cálcio dependentes de interações funcionais.

Watch this Scientific Journal Video about Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes at JoVE.com

Measuring Near Plasma Membrane and Global Intracellular Calcium Dynamics in Astrocytes

Descrevemos como medir perto da membrana e global a dinâmica do cálcio intracelular em astrócitos cultivados utilizando reflexão interna total e microscopia de epifluorescência.

Medição Perto membrana plasmática e global Dynamics cálcio intracelular em astrócitos

The brain contains glial cells.  Astrocytes, a type of glial cell, have long been known to provide a passive supportive role to neurons. However, ...

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