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

Khiroug, S.S., Pryazhnikov, E., Coleman, S.K., Jeromin, A., Keinänen, K. and Khiroug, L. 2009. Dynamic visualization of membrane-inserted fraction of pHluorin-tagged channels using repetitive acidification technique. BMC Neuroscience. 10 141. https://doi.org/10.1186/1471-2202-10-141

TitleDynamic visualization of membrane-inserted fraction of pHluorin-tagged channels using repetitive acidification technique.
TypeJournal article
AuthorsKhiroug, S.S., Pryazhnikov, E., Coleman, S.K., Jeromin, A., Keinänen, K. and Khiroug, L.
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 technique provides a more accurate way of monitoring the PM-inserted fraction of fluorescently tagged molecules and offers a good temporal and spatial resolution.

Article number141
JournalBMC Neuroscience
Journal citation10
ISSN1471-2202
Year2009
PublisherBMC
Publisher's version
License
CC BY 2.0
File Access Level
Open (open metadata and files)
Digital Object Identifier (DOI)https://doi.org/10.1186/1471-2202-10-141
PubMed ID19948025
Web address (URL)http://europepmc.org/abstract/med/19948025
Publication dates
Published30 Nov 2009

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