STOCK Tg(Thy1-Clomeleon)13Gjau/J

Stock number: 013163
Research areas: Neurobiology Research, Research Tools

Description

This thy1::Clomeleon, CLM13, mutant mouse strain is an effective tool for measuring changes of intracellular chloride ion concentration in neurons by fluorescence spectrophotometry, and may be useful in studies requiring synaptic inhibition imaging.

Detailed description

These transgenic mice express Clomeleon, a fluorescent fusion protein containing CFP and topaz YFP that acts as a ratiometric indicator for chloride ions, under the control of the mouse Thy1, thymus cell antigen 1, theta, promoter (Thy1.2). Transgene expression in founder line CLM13 is detected at high levels in the hippocampal dentate gyrus and CA1 region, amygdala, and nuclei and mossy fibers of the cerebellum. No expression is detected in the external plexiform layer, mitral cells or internal plexiform layer of the olfactory bulb, layer 1 of the neocortex, cerebellar granule or purkinje cells, or retinal biopolar and amacrine cells. In the presence of low chloride ion concentrations, YFP fluoresces as a FRET (Fluorescence Resonance Energy Transfer ) acceptor for CFP. As the chloride ion concentration increases, YFP fluorescence decreases and CFP fluorescence increases. Using an appropriate calibration curve, the ratio of YFP and CFP fluorescence peaks indicate [Cl-]. Mice homozygous for the transgenic insert are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. The Donating Investigator reports that homozygotes have a lower reproduction rate than hemizygotes.

Development

A transgenic construct containing Clomeleon, a fluorescent fusion protein containing CFP and topaz YFP, under the control of the mouse Thy1, thymus cell antigen 1, theta, promoter (Thy1.2) was injected into fertilized ICR mouse eggs. Founder line CLM13 was subsequently established. The mice were backcrossed to C57BL/6 for 2 generations. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Tg(Thy1-Clomeleon)13Gjau
Name: transgene insertion 13, George Augustine
MGI accession ID: MGI:4847926
Generation method: Transgenic
Attributes: Reporter
Synonyms: Tg(Thy1-CFP,Topaz)CLM13Gjau; Tg(Thy1-CFP,YFP)CLM13Gjau
Marker symbol: Tg(Thy1-Clomeleon)13Gjau
Marker name: transgene insertion 13, George Augustine
Chromosome: UN
Strain of origin: ICR
Expressed genes: CFP,Cyan Fluorescent Protein,jellyfish; YFP,Yellow Fluorescent Protein,jellyfish; Clomeleon,Clomeleon,
Site of expression: CFP and YFP are expression is dependent on chloride ion concentration in the hippocampal dentate gyrus and CA1 region, amygdala, and nuclei and mossy fibers of the cerebellum.
Molecular note: A transgenic construct containing Clomeleon, a fluorescent fusion protein containing CFP and topaz YFP, under the control of the mouse Thy1, thymus cell antigen 1, theta, promoter (Thy1.2) was injected into fertilized ICR mouse eggs. Founder line CLM13 was subsequently established. Transgenic mice express Clomeleon, a fluorescent fusion protein containing CFP and topaz YFP that acts as a ratiometric indicator for chloride ions, under the control of the mouse Thy1, thymus cell antigen 1, theta, promoter (Thy1.2). Transgene expression in founder line CLM13 is detected at high levels in the hippocampal dentate gyrus and CA1 region, amygdala, and nuclei and mossy fibers of the cerebellum. No expression is detected in the external plexi form layer, mitral cells or internal plexiform layer of the olfactory bulb, layer 1 of the neocortex, cerebellar granule or purkinje cells, or retinal biopolar and amacrine cells. In the presence of low chloride ion concentrations, YFP fluoresces as a FRET (Fluorescence Resonance Energy Transfer ) acceptor for CFP. As the chloride ion concentration increases, YFP fluorescence decreases and CFP fluorescence increases. Using an appropriate calibration curve, the ratio of YFP and CFP fluorescence peaks indicate [Cl-].