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Activation of cerebellar parallel fibers monitored in transgenic mice expressing a fluorescent Ca 2+ indicator protein
Author(s) -
DíezGarcía Javier,
Matsushita Shinichi,
Mutoh Hiroki,
Nakai Junichi,
Ohkura Masamichi,
Yokoyama Jennifer,
Dimitrov Dimitar,
Knöpfel Thomas
Publication year - 2005
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1111/j.1460-9568.2005.04250.x
Subject(s) - antidromic , genetically modified mouse , autofluorescence , stimulation , orthodromic , cerebellum , cerebellar cortex , transgene , green fluorescent protein , fluorescence , chemistry , biophysics , microbiology and biotechnology , neuroscience , biology , biochemistry , physics , quantum mechanics , gene
Genetically encoded fluorescent Ca 2+ indicator proteins (FCIPs) are promising tools to study Ca 2+ signaling in large assemblies of nerve cells. Currently, there are few examples of stable transgenic mouse lines that functionally express such sensors in well‐defined neuronal cell populations. Here we report the generation and characterization of transgenic mice expressing an FCIP under the 5′ regulatory sequences of the Kv3.1 potassium channel promoter. In the cerebellar cortex, expression was restricted to granule cells. We first demonstrated reliable measurements of Ca 2+ transients from beams of parallel fibers and compared the FCIP signals with intrinsic autofluorescence signals. We demonstrate that, in a transgenic line that exhibits a high expression level of the FCIP, autofluorescence signals are negligible and stimulation‐induced fluorescence transients represent FCIP signals. Using frontal cerebellar slices we imaged antidromic activation of granule cells following electrical stimulation of parallel fibers and orthodromic activation of beams of parallel fibers following electrical stimulation of granule cells. We found that paired pulse‐induced presynaptic Ca 2+ transients of parallel fibers are not affected by blockade of N ‐methyl‐ d ‐aspartate receptors.