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

Detta arbete stöddes av Uehara Memorial Foundation i Japan (till ES) samt Whitehall Foundation, National Institute of neurologiska sjukdomar och stroke samt en Stein-Oppenheimer Endowment Award (till BSK).

Experimentella Den experimentella förfarande består av två viktiga delar som beskrivs i ett steg klokt sätt nedan. Del 1: Förbereda hippocampus astrocyternas KULTURER Kortfattat var blandade hippocampus astrocyternas-neuron kulturer prepareras med ett väletablerat protokoll 1,2,3. Vi optimerat proceduren att ge friska odlade astrocyter. Alla förfaranden som anges nedan bör ske i en steril miljö som ett laminärt flöde...

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Det är väl etablerat att astrocyter visa intracellulär kalcium höjder. Dessa uppträda spontant, kan utlösas av nervaktivitet eller genom tillämpning av agonister för att aktivera receptorer på astrocyternas ytan 11. En viktig och kontroversiell fråga är om astrocyternas intracellulär kalcium förhöjningar kan utlösa frisättning av signalmolekyler som aktiverar receptorer på nervceller 11, 12. Detta är kontroversiellt eftersom det har belägg för och emot denna uppfattning, vilket betonas i de översyn...

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measuring near plasma membrane and global intracellular calcium dynamics in astrocytes

Hjärnan innehåller gliaceller. Astrocyter, en typ av stödjeceller celler, har länge varit kända för att ge en passiv stödjande roll till nervceller. Tyder dock allt tydligare att astrocyterna också aktivt kan delta i hjärnans funktion med hjälp av funktionella interaktioner med nervceller. Men många grundläggande aspekter av astrocyternas biologi är fortfarande kontroversiella, otydliga och / eller experimentellt outforskat. En viktig fråga är dynamiken i intracellulära kalcium transienter i astrocyter. Detta är relevant eftersom kalcium är väl etablerad som en viktig andra budbärare och eftersom det har föreslagits att astrocyternas kalcium höjder kan plötsligt frisätter sändare från astrocyter. Dock har det inte funnits någon detaljerad eller tillfredsställande beskrivning av nära plasmamembranet kalcium signalering i astrocyter. Total inre reflektion fluorescens (TIRF) mikroskopi är ett kraftfullt verktyg för att analysera fysiologiskt relevanta signalering händelser inom ca 100 nm i plasmamembranet av levande celler. Här använder vi TIRF mikroskopi och beskriver hur bildskärmen nära plasmamembranet och globala intracellulär dynamik kalcium nästan samtidigt. De ytterligare förfining och systematisk tillämpning av denna metod har potential att informera om de exakta detaljerna i astrocyternas kalcium signalering. En detaljerad förståelse av astrocyternas kalcium dynamik kan ge en grund för att förstå om, hur, när och varför astrocyter och nervceller genomgår kalcium-beroende funktionella interaktioner.

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

Vi beskriver hur man mäter nära membranet och globala intracellulär dynamik kalcium i odlade astrocyter med total inre reflektion och epifluorescence mikroskopi.

Mätning nära plasmamembran och Global Intracellulär Dynamics Kalcium i Astrocyter

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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