ChAT-ChR2-YFP BAC 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, for fluorescent labeling of cholinergic neuronal populations, or for studying the consequences of overactive cholinergic signaling in information processing, memory, behavior and physical fitness.
Guoping Feng, Massachusetts Institute of Technology
Genetic Background | Generation |
---|---|
N6+N3F2
|
Allele Type |
---|
Transgenic (Reporter) |
Starting at:
$255.00 Domestic price for female 4-week |
333.51 Domestic price for breeder pair |
Because the vesicular acetylcholine transporter gene (Slc18a3 or VAChT) within the Chat locus on the BAC transgene was not disrupted, ChAT-mhChR2-YFP BAC transgenic mice also have increased VAChT expression in hippocampus and brainstem. VAChT overexpression results in increased cholinergic tone; the functional consequence of which is increased physical endurance but severe cognitive deficits in attention, working memory and spatial memory.
ChAT-ChR2-YFP BAC transgenic mice have expression of the mhChR2::YFP fusion protein directed to cholinergic neuronal populations by the mouse choline acetyltransferase (Chat or ChAT) promoter/enhancer regions on the BAC transgene. The mhChR2::YFP fusion protein is composed of a mammalian codon-optimized Chlamydomonas reinhardtii-derived channelrhodopsin-2 that was modified to harbor a gain-of-function H134R substitution (mhChR2; also called hChR2-H134R) fused in-frame with an enhanced yellow fluorescent protein (EYFP). The mhChR2 is designed to cause larger stationary photocurrents compared to ChR2. The bacterial opsins are retinal-binding proteins that provide light-dependent ion transport and sensory functions to a family of halophilic bacteria; and this mhChR2 functions as a blue light-driven cation channel that depolarizes the cell and causes action potentials. As such, illuminating mhChR2-expressing neurons with blue light (450-490 nm) leads to rapid and reversible photostimulation of action potential firing/neural activity in these cells. Hemizygous mice are viable and fertile with normal life expectancy, regardless of maternal or paternal inheritance of the transgene.
The donating investigator reports that EYFP expression is visible by direct fluorescence (epifluorescence microscope). ChAT-mhChR2-YFP mice derived from founder line 6 (ChAT-ChR2-YFP line 6) exhibit strong EYFP expression in striatum and basal forebrain, trochlear nucleus, medial habenula, interpeduncular nucleus and various brainstem motor neuron nuclei. Lower EYFP expression is found in cortex, hippocampus, and other brain regions. ChAT co-staining shows precise co-localization with mhChR2-EYFP expression neurons; suggesting mhChR2-EYFP labeled neurons in this line are cholinergic neurons. High EYFP fluorescence is also observed in the ventral gray horn in transverse section of the spinal cord. The donating investigator also reports that these ChAT-mhChR2-YFP line 6 mice exhibit a similar expression pattern as ChAT-mhChR2-YFP line 5 (Stock No. 014545), but line 6 mice have brighter EYFP expression.
Because the vesicular acetylcholine transporter gene (Slc18a3 or VAChT) within the Chat locus on the BAC transgene was not disrupted, VAChT protein is increased more than 5-fold in hippocampus and more than 3-fold in brainstem of ChAT-ChR2-YFP line 6 mice. This VAChT overexpression results in increased cholinergic tone; the functional consequence of which is increased physical endurance but severe cognitive deficits in attention, working memory and spatial memory. ChAT-ChR2-EYFP line 6 mice also consume more food and water during the dark cycle compared to control mice. Compared to wildtype animals, ChAT-ChR2-EYFP line 6 mice show no changes in metabolic profile, locomotor activity in a novel environment, anxiety-like behavior, depression-like behavior, gross sensorimotor function, and ability to use cues to learn a task.
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-ChR2-YFP line 9 (Stock No. 007615),
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),
VGAT-ChR2-YFP line 8 (Stock No. 014548),
and TpH2-ChR2-YFP line 5 (Stock No. 014555).
The ChAT-mhChR2-YFP BAC transgene (or ChAT-ChR2-YFP BAC transgene) was designed in the laboratory of Dr. Guoping Feng (Massachusetts Institute of Technology). First, a channelrhodopsin-2 cDNA sequence derived from green alga Chlamydomonas reinhardtii was modified to harbor codons optimized for mammalian expression and a gain-of-function H134R substitution (CAC to CGC) designed to cause larger stationary photocurrents. This mhChR2 sequence (also called hChR2-H134R) was fused in-frame to the amino terminus of an enhanced yellow fluorescent protein sequence (EYFP) via a Not1 site with GCGGCCGCC linker sequence. This mhChR2::YFP fusion protein (also called hChR2-H134R-EYFP) sequence was inserted into the coding region of the mouse choline acetyltransferase (Chat or ChAT) locus on the RP23-246B12 bacterial artificial chromosome (BAC) via homologous recombination. This was designed to disable transcription from the Chat locus, and none of the other loci on the BAC were altered. The BAC also includes the vesicular acetylcholine transporter gene (Slc18a3 or VAChT) that, because it is located in the intron between the first and second exons of the Chat gene, is under the control of the Chat promoter. The resulting ~220 kb ChAT-mhChR2-YFP BAC transgene was injected into B6SJLF1 fertilized oocytes. Transgenic founders were bred with C57BL/6J to generate the ChAT-mhChR2-YFP line 6 colony (also called ChAT-ChR2-YFP line 6). The colony was backcrossed to C57BL/6J mice for a total of at least five generations prior to sending to The Jackson Laboratory Repository in 2011. Upon arrival, sperm was cryopreserved. To generate the living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J female mice (Stock No. 000664). To maintain our live ChAT-ChR2-YFP line 6 colony, hemizygous females are bred with wildtype (noncarrier) males from the colony or with C57BL/6J inbred males. There are approximately 50 copies of the transgene present. Of note, the donating investigator may not have fixed the Y chromosome to the C57BL/6J genetic background during backcrossing.
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
---|---|
Expressed Gene | COP4, Channelrhodopsin, Chlamydomonas |
Expressed Gene | Slc18a3, solute carrier family 18 (vesicular monoamine), member 3, mouse, laboratory |
Site of Expression | EYFP expression in cholinergic neurons of the striatum, trochlear nucleus, medial habenula, interpeduncular nucleus and various brainstem motor neuron nuclei. Lower EYFP expression is found in cortex, hippocampus, and other brain regions. |
Allele Name | transgene insertion 6, Guoping Feng |
---|---|
Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | ChAT-ChR2-YFP line 6 |
Gene Symbol and Name | Tg(Chat-COP4*H134R/EYFP,Slc18a3)6Gfng, transgene insertion 6, Guoping Feng |
Gene Synonym(s) | |
Promoter | Chat, choline acetyltransferase, mouse, laboratory |
Expressed Gene | YFP, Yellow Fluorescent Protein, jellyfish |
Expressed Gene | COP4, Channelrhodopsin, Chlamydomonas |
Expressed Gene | Slc18a3, solute carrier family 18 (vesicular monoamine), member 3, mouse, laboratory |
Site of Expression | EYFP expression in cholinergic neurons of the striatum, trochlear nucleus, medial habenula, interpeduncular nucleus and various brainstem motor neuron nuclei. Lower EYFP expression is found in cortex, hippocampus, and other brain regions. |
Strain of Origin | (C57BL/6 x CBA)F1 |
Chromosome | UN |
Molecular Note | Transgene expression of the mhChR2::YFP fusion protein is directed to cholinergic neuronal populations by the mouse choline acetyltransferase (Chat or ChAT) promoter/enhancer regions on the BAC transgene. The mhChR2::YFP fusion protein is composed of a mammalian codon-optimized Chlamydomonas reinhardtii-derived channelrhodopsin-2 that was modified to harbor a gain-of-function H134R substitution (mhChR2; also called hChR2-H134R) fused in-frame with an enhanced yellow fluorescent protein (EYFP). The BAC used to generate the construct includes the vesicular acetylcholine transporter gene (Slc18a3 or VAChT). Slc18a3 is located in the intron between the first and second exons of the Chat gene and is under the control of the Chat promoter. VAChT protein is overexpressed in the hippocampus and brainstem. The mhChR2 is designed to cause larger stationary photocurrents compared to ChR2. The bacterial opsins are retinal-binding proteins that provide light-dependent ion transport and sensory functions to a family of halophilic bacteria; and this mhChR2 functions as a blue light-driven cation channel that depolarizes the cell and causes action potentials. As such, illuminating mhChR2-expressing neurons with blue light (450-490 nm) leads to rapid and reversible photostimulation of action potential firing/neural activity in these cells. EYFP expression is visible by direct fluorescence (epifluorescence microscope). ChAT-mhChR2-YFP mice derived from founder line 6 exhibit strong EYFP expression in striatum, trochlear nucleus, medial habenula, interpeduncular nucleus and various brainstem motor neuron nuclei. Lower EYFP expression is found in cortex, hippocampus, and other brain regions. ChAT co-staining shows precise co-localization with mhChR2-EYFP expression neurons; suggesting mhChR2-EYFP labeled neurons in this line are cholinergic neurons. High EYFP fluorescence is also observed in the ventral gray horn in transverse section of the spinal cord. The donating investigator also reports that these ChAT-mhChR2-YFP line 6 mice exhibit a similar expression pattern as ChAT-mhChR2-YFP line 5 (JAX Stock No. 014545), but line 6 mice have brighter EYFP expression. |
Mutations Made By | Guoping Feng, Massachusetts Institute of Technology |
Hemizygous mice are viable and fertile with normal life expectancy, regardless of maternal or paternal inheritance of the transgene. When maintaining a live colony, hemizygous females may be bred with wildtype (noncarrier) males from the colony or with C57BL/6J inbred male mice (Stock No. 000664).
When using the ChAT-ChR2-EYFP line 6 mouse strain in a publication, please cite the originating article(s) and include JAX stock #014546 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Hemizygous or Non carrier for Tg(Chat-COP4*H134R/EYFP)6Gfng |
Frozen Mouse Embryo | B6.Cg-Tg(Chat-COP4*H134R/EYFP Slc18a3)6Gfng/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(Chat-COP4*H134R/EYFP Slc18a3)6Gfng/J | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(Chat-COP4*H134R/EYFP Slc18a3)6Gfng/J | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Tg(Chat-COP4*H134R/EYFP Slc18a3)6Gfng/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.