B6.129(Cg)-Gria2^tm2Rlh/J

Stock number: 024419
Product name: GluR2 K882A KI
Research areas: Neurobiology Research

Description

A K882A mutation was introduced to GluA2/Gria2 to prevent PKCα-mediated phosphorylation at serine 880 (S880). This eliminates cerebellar long-term depression (LTD), reported to be critical for certain types of motor learning.

Detailed description

Cerebellar long-term depression (LTD) is a major form of synaptic plasticity that is thought to be critical for certain types of motor learning. Phosphorylation of GluR2 (Gria2, glutamate receptor, ionotropic, AMPA2 (alpha 2)) on serine-880 has been suggested to contribute to the endocytic removal of postsynaptic AMPA receptors during LTD.

In this mutant strain, a lysine (K) to alanine (A) mutation was introduced to the consensus recognition motif for protein kinase C (PKC) at amino acid 882 to prevent PKCα-mediated phosphorylation at serine 880 (S880) of the GRIA2 protein. Western blots performed with a phospho-specific S880 antibody reveal that basal S880 phosphorylation is intact at levels similar to those observed in wildtype littermates, but PKC-mediated S880 phosphorylation is abolished. The mutation does not interfere with binding to the PDZ domain-containing proteins PICKI and GRIPI. Cerebellar architecture and Purkinje cell morphology appear grossly normal by Nissl and Golgi staining. Cerebellar LTD is absent. Mice are viable and without any observable developmental defects.

 

Development

A lysine (K) to alanine (A) change at amino acid 882 was created in exon 15 (encoding transmembrane domain 4 and the C terminus) of Gria2. This corresponds to an AAA ATT TAG to GCC ATA TGA sequence change (also introducing a BslI site). A loxP-flanked neomycin cassette was introduced to the intron 14 in reverse transcriptional orientation. Linearized targeting vector was electroporated into (129X1/SvJ x 129S1/Sv)F1-Kitl⁺-derived R1 embryonic stem (ES) cells. Resultant chimeric mice were checked for germline transmission and the neomycin cassette was excise by breeding to CMV-Cre mice. Cre was bred out of the line by backcrossing to C57BL/6 for 10 generations.

Allele

Symbol: Gria2^tm2Rlh
Name: targeted mutation 2, Richard L Huganir
MGI accession ID: MGI:3623451
Generation method: Targeted
Attributes: Not Specified
Marker symbol: Gria2
Marker name: glutamate receptor, ionotropic, AMPA2 (alpha 2)
Chromosome: 3
Strain of origin: (129X1/SvJ x 129S1/Sv)F1-Kitl⁺
Molecular note: A targeting vector was designed to insert a floxed neo into the locus as well as the K882A substitution into the exon encoding transmembrane domain 4 and the C terminus region. The neo was subsequently removed via cre recombinase expression. A reduction in the levels of the long isoform was found particularly in mutants of 1 month of age.