STOCK Ehd1^tm1Mhor/J

Stock number: 023073
Research areas: Cell Biology Research, Research Tools

Description

Mouse embryonic fibroblasts (MEFs) from mice homozygous for this targeted mutation exhibit impaired migration, cell spreading, transferrin recycling and reduced cholesterol levels. These mice may be useful in studies of endocytic transport, intracellular trafficking and recycling of membrane proteins, and cholesterol homeostasis.

Detailed description

The protein encoded by the Ehd1 (EH-domain containing 1) gene contains a protein-binding EH (EPS15 homology) domain and has a role in early endocytic membrane fusion and membrane trafficking of recycling endosomes. Mice that are homozygous for this targeted mutation are viable and fertile. No gene product (protein) is detected by Western blot analysis of lung, heart, testis and skeletal muscle from homozygous mice. However, a truncated 64kDa protein is detected by Western blot analysis of brain tissue from homozygous mice. No gene product (mRNA) is detected in primary mouse embryonic fibroblasts (MEFs) from homozygotes by RT-PCR. Ehd1 deficient MEFs plated on fibronectin have impaired migration and cell spreading. MEFs from homozygotes also exhibit impaired transferrin recycling, lower overall levels of beta-1 integrins on the plasma membrane, abnormal focal adhesion formation, and lower cellular cholesterol and reduced lipid droplet size.

Development

A targeting vector designed by Dr. Mia Horowitz (Tel Aviv University) containing a NEO cassette was inserted into exon 3, which resulted in disruption of part of exon 3, and all of exons 4 and 5. The construct was electroporated into 129S6/SvEvTac derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were aggregated with morulae. The resulting chimeric males were bred to 129/SvEv or Swiss Webster females. The mice were then bred to ICR. 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: Ehd1^tm1Mhor
Name: targeted mutation 1, Mia Horowitz
MGI accession ID: MGI:3712350
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Ehd1
Marker name: EH-domain containing 1
Chromosome: 19
Strain of origin: 129S6/SvEvTac
Molecular note: A neomycin resistance cassette was inserted into exon 3 such that it fused to the first 164 bp of that exon and replaced exons 4 and 5.