C57BL/6-Tg(Thy1-SNCA*E35K*E46K*E61K)3798Nuber/J

Stock number: 032799
Product name: 3KL-3798
Research areas: Neurobiology Research

Description

3KL-3798 transgenic mice express human SNCA with the E35K, E46K, and E61K mutations under the direction of the Thy1.2 (Thy1, thymus cell antigen 1, theta) promoter and pSer129 positive inclusions. They are suitable for use in applications related to the study of Parkinson's disease.

Detailed description

These transgenic 3KL-3798 mice express human SNCA with the E35K, E46K, and E61K mutations under the direction of the Thy1.2 (Thy1, thymus cell antigen 1, theta) promoter. At approximately 8 to 10 months of age homozygous transgenic mice exhibit subtle motor impairment, such as abnormal gait, increased time to climb down a pole, and reduced 4 limb wire test endurance. Homozygotes are viable and fertile. Transgenic mice exhibit E-to-K dose-dependent loss of tetramers and gain of monomers in brain neurons. The resultant K-mutant SNCA monomers display increased pSer129-immunoreactivity and association with vesicular membranes, leading to round cytoplasmic inclusions in neuronal somata and fibers. Homozygous mice from founder line 3798 express human SNCA at levels similar to those of the endogenous mouse SNCA. Usage of homozygous mice (3KL+/+) is recommended due to the overall relative low expression level.

Development

Site-directed mutagenesis of full-length human wild-type α-synuclein (SNCA) cDNA was used to introduce the E35K, E46K, and E61K mutations. The modified human SNCA cDNA was used as the source for the Thy1-3K αS transgene, in which the mutant human SNCA is under control of the Thy1.2 (Thy1, thymus cell antigen 1, theta) promoter. The transgene was microinjected into the pronucleus of C57BL/6J one-cell embryos. Founder line 3798 was subsequently established. The 3KL-3798 transgenic mice were backcrossed to C57BL/6 for more than 10 generations. Hemizygotes were intercrossed to generate homozygotes. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony. 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. All of the 43 markers throughout the genome suggested a C57BL/6 background, while three of the 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/6N;C57BL/6J genetic background.

Allele

Symbol: Tg(Thy1-SNCA*E35K*E46K*E61K)3798Nuber
Name: transgene insertion 3798, Silke Nuber
MGI accession ID: MGI:6281748
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Synonyms: 3KL; Thy1-3K alpha S
Marker symbol: Tg(Thy1-SNCA*E35K*E46K*E61K)3798Nuber
Marker name: transgene insertion 3798, Silke Nuber
Chromosome: UN
Strain of origin: C57BL/6J
Molecular note: The transgene contains a Thy1 promoter sequence directing expression of a modified human alpha synuclein cDNA carrying the E35K, E46K and E61K mutations. Founder line 3798 with low overexpression was subsequently established.