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Transgenic mice expressing a pH and Cl – sensing yellow‐fluorescent protein under the control of a potassium channel promoter
Author(s) -
Metzger Friedrich,
RepunteCaigo Vez,
Matsushita Shinichi,
Akemann Walther,
DiezGarcia Javier,
Ho Chi Shun,
Iwasato Takuji,
Grandes Pedro,
Itohara Shigeyoshi,
Joho Rolf H.,
Knöpfel Thomas
Publication year - 2002
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 - 10.1046/j.0953-816x.2001.01837.x
Subject(s) - green fluorescent protein , transgene , promoter , fluorescence , biology , complementary dna , microbiology and biotechnology , ectopic expression , potassium channel , genetically modified mouse , transfection , gene expression , gene , biophysics , chemistry , biochemistry , physics , quantum mechanics
During the last few years a variety of genetically encodable optical probes that monitor physiological parameters such as local pH, Ca 2+ , Cl – , or transmembrane voltage have been developed. These sensors are based on variants of green‐fluorescent protein (GFP) and can be synthesized by mammalian cells after transfection with cDNA. To use these sensor proteins in intact brain tissue, specific promoters are needed that drive protein expression at a sufficiently high expression level in distinct neuronal subpopulations. Here we investigated whether the promoter sequence of a particular potassium channel may be useful for this purpose. We produced transgenic mouse lines carrying the gene for enhanced yellow‐fluorescent protein (EYFP), a yellow‐green pH‐ and Cl – sensitive variant of GFP, under control of the Kv3.1 K + channel promoter (pKv3.1). Transgenic mouse lines displayed high levels of EYFP expression, identified by confocal microscopy, in adult cerebellar granule cells, interneurons of the cerebral cortex, and in neurons of hippocampus and thalamus. Furthermore, using living cerebellar slices we demonstrate that expression levels of EYFP are sufficient to report intracellular pH and Cl – concentration using imaging techniques and conditions analogous to those used with conventional ion‐sensitive dyes. We conclude that transgenic mice expressing GFP‐derived sensors under the control of cell‐type specific promoters, provide a unique opportunity for functional characterization of defined subsets of neurons.