B6(Cg)-Caly^tm1.1Cber/J

Stock number: 023129
Product name: CalyKO
Research areas: Neurobiology Research, Research Tools

Description

Agonist-stimulated endocytosis of synaptic AMPA glutamate receptors is impaired in mice homozygous for the Caly knockout allele. These mice may be useful in studies of clathrin-mediated endocytosis in the central nervous system, neuronal synaptic vesicle trafficking and recycling, as well as neuregulin processing and AP-3 dependent axonal transport.

Detailed description

Calcyon neuron-specific vesicular protein regulates clathrin-mediated endocytosis and AP-3 dependent endosomal trafficking. These knock-out mice carry a targeted mutation of the Caly gene in which exons 3 through 6 have been excised. Agonist-stimulated endocytosis of synaptic AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) glutamate receptors (AMPARs), proteolytic processing of neuregulin 1, and axonal targeting of AP-3 cargos are impaired in the brains of homozygotes. Cultured hippocampal neurons isolated from homozygotes and CA1 region brain slices lack long-term synaptic depression (LTD). Mice that are homozygous for the targeted mutation are viable and fertile. No gene product (mRNA) is detected by RT-PCR analysis of brain tissue from homozygotes. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background.

Development

A targeting vector was used to insert an FRT site-flanked NEO cassette into intron 6 and loxP sites upstream of exon 3 and downstream of exon 6. The construct was electroporated into unspecified C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells, containing the Cal^fl allele, were injected into BALB/c blastocysts. The resulting male chimeric animals were crossed to C57BL/6 female mice. Heterozygous Calcyon^fl mice were crossed to generate homozygotes. Homozygotes were crossed to CMV^CRE mice to excise exons 3 through 6 and the FRT-flanked NEO cassette. The mice were then backcrossed to C57BL/6 for at least 10 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6 genetic background. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Caly^tm1.1Cber
Name: targeted mutation 1.1, Clare Bergson
MGI accession ID: MGI:3838019
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Caly
Marker name: calcyon neuron-specific vesicular protein
Chromosome: 7
Strain of origin: C57BL/6
Molecular note: Mice homozygous for the floxed Caly^tm1Cber allele were crossed with Tg(CMV-cre)1Cgn transgenic mice to generate null homozygotes that were then used for further breeding. No mRNA was detectable by RT-PCR on cortex and hind-brain extracts from homozygotes.