Dynamic visualization of membrane-inserted fraction of pHluorin-tagged channels using repetitive acidification technique.

ABSTRACT:

Background

Changes in neuronal excitability, synaptic efficacy and generally in cell signaling often result from insertion of key molecules into plasma membrane (PM). Many of the techniques used for monitoring PM insertion lack either spatial or temporal resolution.

Results

We improved the imaging method based on time-lapse total internal reflection fluorescence (TIRF) microscopy and pHluorin tagging by supplementing it with a repetitive extracellular acidification protocol. We illustrate the applicability of this method by showing that brief activation of NMDA receptors ("chemical LTP") in cultured hippocampal neurons induced a persistent PM insertion of glutamate receptors containing the pHluorin-tagged GluR-A(flip) subunits.

Conclusion

The repetitive acidification

SUBMITTER: Khiroug SS 

PROVIDER: S-EPMC2794868 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature