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J Neurosci. 2011 Feb 23;31(8):2855-67. doi: 10.1523/JNEUROSCI.6064-10.2011.

Imaging light responses of targeted neuron populations in the rodent retina.

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  • 1Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia 20147, USA. borghuisb@janelia.hhmi.org

Abstract

Decoding the wiring diagram of the retina requires simultaneous observation of activity in identified neuron populations. Available recording methods are limited in their scope: electrodes can access only a small fraction of neurons at once, whereas synthetic fluorescent indicator dyes label tissue indiscriminately. Here, we describe a method for studying retinal circuitry at cellular and subcellular levels combining two-photon microscopy and a genetically encoded calcium indicator. Using specific viral and promoter constructs to drive expression of GCaMP3, we labeled all five major neuron classes in the adult mouse retina. Stimulus-evoked GCaMP3 responses as imaged by two-photon microscopy permitted functional cell type annotation. Fluorescence responses were similar to those measured with the small molecule dye OGB-1. Fluorescence intensity correlated linearly with spike rates >10 spikes/s, and a significant change in fluorescence always reflected a significant change in spike firing rate. GCaMP3 expression had no apparent effect on neuronal function. Imaging at subcellular resolution showed compartment-specific calcium dynamics in multiple identified cell types.

PMID:
21414907
[PubMed - indexed for MEDLINE]
PMCID:
PMC3521507
Free PMC Article
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