B6.Cg-Tg(Thy1-COP4/EYFP)9Gfng/J

Stock number: 007615
Product name: Thy1-ChR2-YFP
Research areas: Neurobiology Research, Research Tools

Description

Tg(Thy1-COP4/EYFP)9Gfng (or Thy1-ChR2-YFP) transgenic mice may be useful in optogenetic studies for rapid control of motor behavior by addition or removal of light, for ex vivo and in vivo studies of neural circuitry/connectivity following illumination, and for fluorescent labeling of Thy1-expressing cells throughout the brain (including cortex, hippocampus, thalamus, midbrain, brainstem, cerebellar mossy fibers and retinal ganglion cells).

Detailed description

These Thy1-ChR2-YFP founder line 9 transgenic mice express the light-activated ion channel, Channelrhodopsin-2 (from the green alga Chlamydomonas reinhardtii), fused to Yellow Fluorescent Protein (ChR2-YFP) under the control of the mouse thymus cell antigen 1 (Thy1) promoter. Hemizygotes are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Expression of the transgenic ChR2-YFP fusion protein is detected throughout the brain, including in the cortex, hippocampus, thalamus, midbrain, brainstem, cerebellar mossy fibers and retinal ganglion cells. Neurons expressing the transgene are morphologically and physiologically comparable to non-mutant neurons. This mutant mouse strain may be useful for ex vivo and in vivo neural circuitry mapping studies using light stimulation.

The ChR2-YFP fusion protein is composed of a Chlamydomonas reinhardtii-derived channelrhodopsin-2 (ChR2) fused in-frame with an enhanced yellow fluorescent protein (EYFP).

The bacterial opsins are retinal-binding proteins that combine a light-sensitive domain with an ion channel or pump; providing light-dependent ion transport, membrane potential alteration, and sensory functions to bacteria. ChR2 functions as a blue light-driven cation channel that depolarizes the cell and causes action potentials. As such, illuminating ChR2-expressing neurons with blue light (~470 nm) leads to rapid and reversible photostimulation of action potential firing/neural activity in these cells.

This optogenetic strain is one of many from the same transgene creator/donating investigator with light-inducible neurobiology applications; including Thy1-ChR2-YFP line 18 (Stock No. 007612), Thy1-eNpHR-YFP line 2 (Stock No. 012332), Thy1-eNpHR-YFP line 4 (Stock No. 012334), Thy1-vChR1-YFP line 1 (Stock No. 012341), Thy1-vChR1-YFP line 4 (Stock No. 012344), Thy1-vChR1-YFP line 8 (Stock No. 012348), Thy1-mhChR2-YFP Line 20 (Stock No. 012350), Prv-mhChR2-YFP Line 15 (Stock No. 012355), ChAT-ChR2-YFP line 5 (Stock No. 014545), ChAT-ChR2-YFP line 6 (Stock No. 014546), VGAT-ChR2-YFP line 8 (Stock No. 014548), and TpH2-ChR2-YFP line 5 (Stock No. 014555).

View YFP fluorescence in sagittal brain sections for this strain.

Development

A transgenic construct containing Channelrhodopsin-2 from the green alga Chlamydomonas reinhardtii and Yellow Fluorescent Protein fusion gene under the control of the mouse thymus cell antigen 1 promoter was injected into C57BL/6 X SJL F1 hybrid donor eggs. Founder line 9 was subsequently established. The mice were backcrossed to C57BL/6 for six generations before arriving at The Jackson Laboratory.

Allele

Symbol: Tg(Thy1-COP4/EYFP)9Gfng
Name: transgene insertion 9, Guoping Feng
MGI accession ID: MGI:3719563
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Thy1-COP4/EYFP)9Gfng
Marker name: transgene insertion 9, Guoping Feng
Chromosome: UN
Strain of origin: C57BL/6 x SJL
Expressed genes: YFP,Yellow Fluorescent Protein,jellyfish; COP4,Channelrhodopsin,Chlamydomonas
Site of expression: widespread expression in the brain including the cortex, hippocampus, thalamus, midbrain, brainstem, cerebellar mossy fibers amd retinal ganglion cells; channelrhodopsin-2 and YFP
Molecular note: This transgene gives rise to a fusion protein composed of the first 315 amino acids of the light-activated, non-specific ion channel chlamyopsin 4 (COP4), also known as channelopsin 2 (CHOP2) and as channelrhodopsin 2 (ChR2), from the green alga Chlamydomonas reinhardtii joined to enhanced yellow fluorescent protein. The chimeric cDNA has replaced the Thy1 coding sequence and its associated introns, which contain sequences directing expression in non-neuronal cell types. The vector contains the entire Thy1 gene (including the essential promoter and neuron-specific upstream regulatory sequences, 3' polyA signal sequences and neuronal-specific expression signal containing introns) with the coding region removed (from the ATG in exon 2 to the end in exon 4, including intervening introns 2 and 3).

Immunohistochemical analysis shows that mice with this transgene express the chimeric protein in neurons of several brain regions, including various nuclei of the brainstem, midbrain and thalamus; CA1 and CA3 pyramidal neurons of the hippocampus; strongly in layer 5 neurons of the cortex; and in retinal ganglion cells.
General note: Lines initiated from 7 of 10 transgenic founder mice exhibited expression of the chimeric protein in the central and/or peripheral nervous system, the specific pattern of expression varying among lines.