B6.FVB(Cg)-Tg(Chat-EGFP/Rpl10a,Slc18a3)DW167Htz/J

Stock number: 030250
Product name: Chat-EGFP/Rpl10a
Research areas: Neurobiology Research, Research Tools

Description

This bacTRAP transgenic strain, incorporating an EGFP-L10a ribosomal fusion protein targeted to the Chat gene, enables cell labeling, sorting, and affinity purification of cell-specific polysomal RNAs. The BAC also overexpresses Slc18a3 and mice show altered sensitivity to nicotine as well as increased stereotypic behavior to other drugs of abuse.

Detailed description

This bacTRAP line incorporates an EGFP-L10a ribosomal fusion protein targeted to the translational start site of the Chat (choline acetyltransferase) gene. The fusion protein enables affinity purification of CNS cell-specific polysomal RNAs which can be used for gene expression profiling.

Expression was carefully assayed by immunohistochemistry (IHC) with an antibody against enhanced green fluorescent protein (EGFP). The cerebellum, spinal cord, basal forebrain and corpus striatum, brainstem, and cerebral cortex were surveyed. Primary labeling was found to occur in motor neurons and cholinergic interneurons of the spinal cord, cholinergic neurons of the corpus striatum, cholinergic projection neurons of the basal forebrain, and motor neurons and midbrain cholinergic neurons of the brain stem. Fluorescence can also be visualized directly.

Images showing neuron, large multipolar interneuron, and motor neuron expression patterns may be found on the Gensat TRAP cell Showcase and Gensat gene expression pages.

These BAC transgenic mice also overexpress Slc18a3 (solute carrier family 18 (vesicular monoamine), member 3; also called VAChT). No unusual behaviors have been noted.

Development

A shuttle vector was used to introduce sequences encoding an EGFP-L10a ribosomal fusion protein into the translation start site of the Chat gene contained within BAC RP23-431D9. The modified BAC was introduced to FVB embryos. Founders were bred to Swiss Webster mice, then subsequently backcrossed to C57BL/6J mice for 9 generations by the donating laboratory. This is founder line DW167.

Allele

Symbol: Tg(Chat-EGFP/Rpl10a,Slc18a3)DW167Htz
Name: transgene insertion DW167, Nathaniel Heintz
MGI accession ID: MGI:5496680
Generation method: Transgenic
Attributes: Reporter
Synonyms: ChAT-DW167; Tg(Chat-EGFP/Rpl10a)DW167Htz
Marker symbol: Tg(Chat-EGFP/Rpl10a,Slc18a3)DW167Htz
Marker name: transgene insertion DW167, Nathaniel Heintz
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: GFP,Green Fluorescent Protein,; Rpl10a,ribosomal protein L10A,mouse, laboratory
Site of expression: EGFP is expressed primarily in motor neurons and cholinergic interneurons of the spinal cord, cholinergic neurons of the corpus striatum, cholinergic projection neurons of the basal forebrain, and motor neurons and midbrain cholinergic neurons of the brain stem.
Molecular note: Transgenic mice were generated using modified BAC Rp23-431D9. EGFP fused to the ribosomal protein L10a was inserted in the BAC in such a way that expression was driven by Chat. 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.