B6.129S4(FVB)-Grin2a^tm1.1Jpleo/J

Stock number: 027998
Product name: Grin2aΔPKC
Research areas: Neurobiology Research

Description

Grin2aΔPKC Grin2a mutant mice have amino acid substitutions that result in nonphosphorylatable PKC sites. These mice may be suitable for use in studies related to synaptic plasticity, memory and learning.

Detailed description

The targeted Grin2a gene encodes the GluN2A NMDA receptor subunit. Mutations in this gene have been associated with Focal Epilepsy and Speech Disorder with or without Mental Retardation and Landau-Kleffner Syndrome. These Grin2aΔPKC mice carry the unphosphorylatable amino acid substitutions S1991A, Y1292F, S1312A and Y1472F (encoded by exon 12), which are sites of Protein kinase C phosphorylation.

Mutant homozygous mice exhibit decreased anxiety-related behaviors, as assessed in open field test, light dark emergence test, and elevated plus maze. Slight spatial working memory impairment is also observed in homozygotes (reduced spontaneous Y maze alternation and reduced delayed T maze alternation). Mice that are homozygous for the targeted mutation are viable and fertile. Gene product (mRNA or protein) expression levels in mutant mice are similar to that observed in wildtype controls, as determined by Northern or Western blot analysis.

Development

A targeting vector containing mutations that result in amino acid substitutions S1991A, Y1292F, S1312A and Y1472F, and a loxP site flanked NEO cassette was used to disrupt exon 12. The construct was electroporated into 129S4/SvJae derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were bred to C57BL/6 mice to achieve germline transmission. The mice were then crossed to Ella-cre mice on the C57BL/6 and FVB genetic background to remove the floxed NEO cassette. Mice that no longer carried the NEO cassette were bred to C57BL/6J for 5 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 4 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.

Allele

Symbol: Grin2a^tm1.1Jpleo
Name: targeted mutation 1.1, John P Leonard
MGI accession ID: MGI:5811328
Generation method: Targeted
Attributes: Not Applicable
Marker symbol: Grin2a
Marker name: glutamate receptor, ionotropic, NMDA2A (epsilon 1)
Chromosome: 16
Strain of origin: 129S4/SvJae
Molecular note: A targeting vector containing mutations that result in amino acid substitutions S1991A, Y1292F, S1312A and Y1472F, and a loxP site flanked NEO cassette was used to disrupt exon 12. Cre-mediated recombination removed the floxed neo cassette.