FVB;129-Pink1^tm1Aub Tg(Prnp-SNCA*A53T)AAub/J

Stock number: 017678
Product name: Pink1^-;PrPmtA
Research areas: Neurobiology Research

Description

This double mutant strain expresses mutant human A53T alpha-synuclein (SNCA) and carries a targeted mutation for Pink1, PTEN induced putative kinase 1, and has applications in studies of the early stages of Parkinson's Disease.

Detailed description

These double mutant mice overexpress the mutant A53T human SNCA, synuclein, alpha (non A4 component of amyloid precursor), and are deficient in endogenous mouse Pink1, PTEN induced putative kinase 1. Mice that are homozygous for both the transgene and the Pink1^tm1Aub allele develop a behavior phenotype at 3 months of age, consisting of impaired spontaneous locomotion in open field tests for horizontal and vertical activity. This behaviour deficit appears much later also in the single mutant mice. In addition, the brain transcriptome profile in striatum of homozygous double mutants shows many consistent early alterations in the pathways of trophic signaling to mitochondria and apoptosis, which are not apparent in the transcriptome profiles of single mutants.

Development

For the Pink1^tm1Aub mutant strain, a targeting construct containing a nucleotide substitution from a G to an A at nucleotide 8343 [encoding G308D; equivalent to the human G309D loss-of-function mutation found in early-onset PARK6-linked Parkinson's Disease (PD)] and a loxP-flanked NEO cassette was targeted to exon 5 of the mouse PTEN induced putative kinase 1 locus (Pink1) via electroporation into a unspecified 129/SvEv-derived embryonic stem (ES) cell line. Correctly targeted ES cells were injected into blastocysts. The colony was maintained on a 129/SvEv background.

For the Tg(Prnp-SNCA*A53T)AAub transgenic strain, a transgenic construct containing the entire translated portion of the mutant A53T human alpha-synuclein (SNCA) from position 22 to 515, driven by an ~3.5 kb fragment of the mouse prion protein promoter (Prnp), and polyadenylation signal sequence, was injected into fertilized FVB/N mouse eggs. Founder line A was subsequently established.

The strains were then intercrossed to generate double mutant mice that were heterozygous for the Pink1^tm1Aub allele and Tg(Prnp-SNCA*A53T)AAub transgene. The double heterozygotes were then crossed to generate mice homozygous for both mutations. The mice were maintained on a mixed 129;FVB background. Upon arrival at The Jackson Laboratory, the mice were crossed to FVB/NJ (Stock No. 001800) at least once to establish the colony.

Allele

Symbol: Tg(Prnp-SNCA*A53T)AAub
Name: transgene insertion A, Georg Auburger
MGI accession ID: MGI:3723258
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(Prnp-SNCA*A53T)AAub
Marker name: transgene insertion A, Georg Auburger
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: SNCA,synuclein alpha,human
Molecular note: A construct containing the entire translated portion of human alpha-synuclein, SNCA from position 22 to 515, driven by an ~3.5 kb fragment of the mouse prion protein promoter, Prnp, was generated. The SNCA cDNA was followed by polyadenylation sequences derived from the 3' untranslated region of the boving growth hormone gene. This construct was microinjected into pronuclei of fertilized FVB/N ova. Transgenic protein expression was detected in the neuropil, with, more rarely, additional protein within neuronal cell bodies and neurites in the brain and motoneurons in the spinal cord. Two lines, A and B, were generated, have both been extensively studied, showing extremely similar phenotypes (J:86222, J:124976).

Allele

Symbol: Pink1^tm1Aub
Name: targeted mutation 1, Georg Auburger
MGI accession ID: MGI:3850370
Generation method: Targeted
Attributes: Null/Knockout; Humanized sequence
Marker symbol: Pink1
Marker name: PTEN induced putative kinase 1
Chromosome: 4
Strain of origin: 129
Molecular note: A p.G308D (g8343a) mutation was created in exon 4 and a floxed neo cassette in inverse orientation was inserted into intron 5 via homologous recombination. The mutation is the equivalent of the G309D mutation found in Parkinson's Disease patients. Northern blot and saturation RT-PCR of splice boundaries in brain and liver mRNA analysis confirmed the absence of full length transcript expression. Saturation RT-PCR also suggests the residual expression of low levels of a large mutant mRNA.