STOCK Ppif^tm1.1Mmos/J

Stock number: 005740
Product name: CyP-D KO
Research areas: Cell Biology Research, Metabolism Research, Research Tools

Description

Mitochondria from these Ppif knock-out mice exhibit an increased capacity to retain calcium and fail to swell/rupture in response to CaCl₂. They are suitable for use in applications related to studies investigating oxidative stress- and ischemia–induced cell death.

Detailed description

Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. Brain architecture and cerebrovasculature are normal. No gene product (protein) is detected by immunoblot analysis of mitochondria isolated from liver tissue of mutant mice. Mitochondria from mutant mice have an increased capacity to retain calcium and fail to swell/rupture in response to CaCl₂, suggesting abnormal permeability transition pore (PTP) function. Mouse embryonic fibroblasts (MEFs) derived from mutant mice are less susceptible to oxidative stress induced in vitro by exposure to hydrogen peroxide than MEFs derived from wildtype mice. In a model of ischemic brain injury employing a middle cerebral artery occlusion protocol, mutant mice exhibited a reduced infarct volume (37% in heterozygotes, and 62% in homozygotes) when compared to wildtype mice. This mutant mouse strain may be useful in studies investigating oxidative stress- and ischemia–induced cell death.

Development

A targeting vector containing a loxP-flanked neomycin resistance gene cassette preceding exons 3-5 and a third loxP site downstream of exon 5 was electroporated into 129X1/SvJ-derived RW4 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a pMC-Cre gene to delete the neo gene (resulting in loxP-flanked exons 3-5 [CypD^flox]) and then injected into C57BL/6 blastocysts. Chimeric mice were crossed to C57BL/6. The resulting heterozygotes were next crossed to FVB/N-Tg(EIIa-cre)C5379Lmgd/J (Stock No. 003314). Mice having been recombined such that exons 3-5 are replaced with a single loxP site (CypD^null) were intercrossed to generate mice homozygous for the deletion.

Allele

Symbol: Ppif^tm1.1Mmos
Name: targeted mutation 1.1, Michael A Moskowitz
MGI accession ID: MGI:3603458
Generation method: Targeted
Attributes: Null/Knockout
Synonyms: CypD^null
Marker symbol: Ppif
Marker name: peptidylprolyl isomerase F (cyclophilin F)
Marker synonyms: CYP3; CypD; Cyp-D; CyP-F; CyP-M; mitochondrial Cyclophilin D; PPIase
Chromosome: 14
Strain of origin: 129X1/SvJ
Molecular note: A targeting vector was designed to insert loxP sites around exons 3-5. Crossing with mice expressing cre in the germ line resulted in a null allele, as was demonstrated by immunoblot of liver samples.