129S1/SvImJ-Trpc4^tm1.1Clph/J

Stock number: 030802
Product name: TRPC4^-
Research areas: Neurobiology Research

Description

Exon 4 of the mouse Trpc4 gene has been excised to create a knock-out allele in this targeted mutant strain. Homozygous mice exhibit diminished innate fear levels in response to innately aversive stimuli.

Detailed description

Exon 4 of the mouse Trpc4 (transient receptor potential cation channel, subfamily C, member 4) gene has been excised to create a knock-out allele in this targeted mutant strain. TRPC4 protein is not detected by Western blot in immunoprecipitation assays in brain microsomes from homozygous knock-out mice.

Homozygous mice exhibit diminished innate fear levels in response to aversive stimuli. They enter the open arms of the elevated plus-maze more frequently and spend more time there than control mice. This indicates homozygous null mice are less anxious than their control littermates. When placed in a well-lit, novel open field, homozygotes spend significantly more time in the anxiogenic center of the arena and enter it more frequently, demonstrating that anxiolytic-like behavioral modifications in mice lacking TRPC4 are more readily revealed under anxiety-provoking conditions. The mice do not exhibit general behavioral deficits reflecting anxiety levels unrelated to changes in behavioral responses. They do not exhibit impairment in the acoustic startle reflex, or in the tail suspension test (a measure of behavioral despair).

Development

A loxP site was introduced to intron 3 and a FRT-flanked PGK-neomycin cassette and loxP site were placed in intron 4 via homologous recombination in 129S1/SvImJ-derived embryonic stem (ES) cells. ES cells harboring the targeting construct were transfected with pOG231, a plasmid for transient cre expression, to excise exon 4 and the neomycin cassette to create a constitutive knock-out allele. ES cells were injected into C57BL/6 mouse blastocysts. Chimeric animals were crossed with 129S1/SvImJ for 10 generations by the donating laboratory.

Excision of exon 4 (encoding amino acids 299-412 within the putative third transmembrane domain) introduced both a frame shift and premature stop codon after the deleted segment.

Allele

Symbol: Trpc4^tm1.1Clph
Name: targeted mutation 1.1, David E Clapham
MGI accession ID: MGI:5615468
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Trpc4
Marker name: transient receptor potential cation channel, subfamily C, member 4
Marker synonyms: CCE1; HTRP4; HTRP-4; STRPC4; TRP4; Trrp4
Chromosome: 3
Strain of origin: 129S1/SvImJ
Molecular note: A loxP site was inserted upstream of exon 4 and an FRT flanked neo cassette and loxP site were inserted downstream of exon 4 via homologous recombination. Cre mediated recombination removed exon 4 and the neo cassette. Western blot analysis confirmed the absence of protein expression in brain microsomes from homozygous mice.