Work

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

Year: 2009

Type: article

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.

Source: BMC Neuroscience

DOI: 10.1186/1471-2202-10-141

Authors: Serguei S Khiroug, Evgeny Pryazhnikov, Sarah K Coleman, Andreas Jeromin, Kari Keinänen, Leonard Khiroug

Institutions: University of Helsinki, Allen Institute for Brain Science

Language: English

Topic: Neuroscience and Neuropharmacology Research

Subfield: Cellular and Molecular Neuroscience

Field: Neuroscience

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,990

Funders: Academy of Finland, Kansainvälisen Liikkuvuuden ja Yhteistyön Keskus