Activation of cerebellar parallel fibers monitored in transgenic mice expressing a fluorescent Ca2+ indicator protein

Javier Díez-García, Shinichi Matsushita, Hiroki Mutoh, Junichi Nakai, Masamichi Ohkura, Jennifer Yokoyama, Dimitar Dimitrov, Thomas Knöpfel

Research output: Contribution to journalArticle

95 Citations (Scopus)

Abstract

Genetically encoded fluorescent Ca2+ indicator proteins (FCIPs) are promising tools to study Ca2+ 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 Ca2+ 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 Ca2+ transients of parallel fibers are not affected by blockade of N-methyl-D-aspartate receptors.

Original languageEnglish
Pages (from-to)627-635
Number of pages9
JournalEuropean Journal of Neuroscience
Volume22
Issue number3
DOIs
Publication statusPublished - 2005 Aug 1
Externally publishedYes

Keywords

  • Green fluorescent protein
  • In vitro
  • Optical imaging
  • Parallel fibers

ASJC Scopus subject areas

  • Neuroscience(all)

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