These mutant mice express the D620N mutation in exon 15 of the Vps35 (VPS35 retromer complex component) gene. VPS35 D620N KI mice recapitulate many of the characteristics of Parkinson's disease including neurodegeneration of DA neurons, accumulation of α-synuclein in DA neurons and age-dependent progressive motor deficits.
Vps35 (VPS35 retromer complex component) encodes a component of the heteropentameric retromer complex, which mediates retrograde transport of cargo proteins from endosomes to the trans-Golgi network, endosome to plasma membrane and mitochondria to peroxisomes or lysome. Vsp35 is highly expressed in dopaminergic neurons. The D620N mutation is associated with late-onset autosomal dominant Parkinson’s disease (PD). The mutation alters cargo recognition of substrates and causes alterations in trafficking. VPS35 D620N knock-in (KI) mice are viable and fertile. Young mice (6-10 months) exhibit no obvious phenotype. However, by 14 months of age homozygotes develop age-dependent progressive motor deficits (reduced travel distance, reduced speed in beam walking), reduced levels of dopamine in the striatum, loss of dopaminergic neurons in the substantia nigra and striatum, increased neuroinflammation, and accumulation and aggregation of α-synuclein in dopaminergic neurons, mitochondrial fragmentation, and increased susceptibility of dopaminergic neurons to environmental toxins. These mice may be useful in studies of Parkinson's disease.
Please note: A strain with inducible expression of D620N is also available, Stock No.021807 B6.Cg-Vps35tm1.2Mjff/J.
A targeted vector containing a floxed "mini-cDNA" (splice acceptor, sequence encoding exons 15 through 17, polyadenylation signal), with a FRT site flanked NEO cassette on the 3'end of the mini-cDNA and a third loxP site, was inserted into intron 14-15. The construct was electroporated into C57BL/6 derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were then crossed to FLP expressing mice on the C57BL/6J genetic background, to remove the NEO cassette. The FLP recombinase transgene was then bred out from the line. Sequencing confirmed the presence of the point mutation.The mice were maintained on the C57BL/6 background. Upon arrival at The Jackson Laboratory, the mice (Stock No. 021807) were crossed to C57BL/6J (Stock No. 000664) at least once. The (Stock No. 021807) mice were then crossed to Cre deleter line on the B6 background (Stock No. 008454) to remove the floxed "mini-cDNA" sequence and allow expression of the D620N mutation. The mice were then bred to remove the Cre recombinase allele.
|Allele Name||targeted mutation 1.1,The Michael J Fox Foundation|
|Allele Type||Targeted (Humanized sequence)|
|Allele Synonym(s)||D620N VPS35 KI; VKI; VPS35D620N|
|Gene Symbol and Name||Vps35, VPS35 retromer complex component|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||A targeted vector containing a floxed "mini-cDNA" including a splice acceptor and normal cDNA sequence for exons 15-17, followed by a frt-flanked neo cassette and a third loxP site, inserted into intron 14, and a nucleotide change causing a D620N mutation in the inherent mouse gene sequence 3' of the insertion. The polyadenylation signal sequence at the end of the mini-cDNA prevents expression of the D620N mutation. Flp-mediated recombination removed the neo cassette. Cre-mediated recombination removed the "mini-cDNA" sequence and allows expression of the D620N mutation.|
Heterozygotes and homozygotes are viable and fertile.
When using the VPS35 D620N
mouse strain in a publication, please cite the originating article(s) and include JAX stock #023409 in your Materials and Methods section.