Characterization and subcellular targeting of GCaMP-type genetically-encoded calcium indicators

Mao, T., O'Connor, D. H., Scheuss, V., Nakai, J., Svoboda, K. (March 2008) Characterization and subcellular targeting of GCaMP-type genetically-encoded calcium indicators. PLoS ONE, 3 (3). e1796.

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URL: http://www.ncbi.nlm.nih.gov/pubmed/18350138

DOI: 10.1371/journal.pone.0001796

Abstract

Genetically-encoded calcium indicators (GECIs) hold the promise of monitoring [Ca(2+)] in selected populations of neurons and in specific cellular compartments. Relating GECI fluorescence to neuronal activity requires quantitative characterization. We have characterized a promising new genetically-encoded calcium indicator-GCaMP2-in mammalian pyramidal neurons. Fluorescence changes in response to single action potentials (17+/-10% DeltaF/F [mean+/-SD]) could be detected in some, but not all, neurons. Trains of high-frequency action potentials yielded robust responses (302+/-50% for trains of 40 action potentials at 83 Hz). Responses were similar in acute brain slices from in utero electroporated mice, indicating that long-term expression did not interfere with GCaMP2 function. Membrane-targeted versions of GCaMP2 did not yield larger signals than their non-targeted counterparts. We further targeted GCaMP2 to dendritic spines to monitor Ca(2+) accumulations evoked by activation of synaptic NMDA receptors. We observed robust DeltaF/F responses (range: 37%-264%) to single spine uncaging stimuli that were correlated with NMDA receptor currents measured through a somatic patch pipette. One major drawback of GCaMP2 was its low baseline fluorescence. Our results show that GCaMP2 is improved from the previous versions of GCaMP and may be suited to detect bursts of high-frequency action potentials and synaptic currents in vivo.

Item Type:	Paper
Subjects:	organs, tissues, organelles, cell types and functions > sub-cellular tissues: types and functions > calcium channel organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions > cell types organs, tissues, organelles, cell types and functions > cell types and functions organs, tissues, organelles, cell types and functions > cell types and functions > cell types > dendritic cells > dendritic spines organs, tissues, organelles, cell types and functions > cell types and functions > cell types > dendritic cells > dendritic spines organs, tissues, organelles, cell types and functions > cell types and functions > cell types > dendritic cells > dendritic spines organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons organs, tissues, organelles, cell types and functions organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > pyramidal neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > pyramidal neurons organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons > pyramidal neurons organs, tissues, organelles, cell types and functions > sub-cellular tissues: types and functions
CSHL Authors:	Mao, Tianyi Scheuss, Volker Svoboda, Karel O'Connor, Daniel H.
Communities:	CSHL labs > Svoboda lab
Depositing User:	Matt Covey
Date:	19 March 2008
Date Deposited:	28 Feb 2013 16:33
Last Modified:	28 Feb 2013 16:33
PMCID:	PMC2262942
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/27499

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