PKCδ- KO mice lack the initiation codon in exon 2 of the protein kinase C, delta gene. This strain may be useful for studying the role of PKCδ in neutrophil recruitment during reperfusion after ischemic injury.
Robert O. Messing, The University of Texas at Austin
PKCδ--KO mice lack exon 2 resulting in loss of expression. PKCδ has been shown to play a role in cardiac ischemia and reperfusion by stimulating the infiltration of neutrophils to the ischemic tissues. Neutrophils release free radicals, enzymes, and cytokines to promote further recruitment of neutrophils and other leukocytes to the site of injury. These inflammatory responses lead to endothelial damage to the effected tissue. PKCδ--KO mice exhibit reduced reperfusion injury following transient ischemia. They show a 70% reduction in stroke size compared with WT mice due to impaired neutrophil function and reduced neutrophil migration into ischemic tissue. They also have impaired neutrophil adhesion, migration, respiratory burst, and degranulation in vitro. Mice contain 80% fewer neutrophils in the cerebral cortex and 65% fewer neutrophils in the striatum, with no difference seen in the peripheral blood.
In a model of proteinuria, albumin overload induces apoptosis in renal tubules, which is less severe in PKCδ- mice than in controls. PKCδ is also important for acute responses to ethanol, since compared to wild type mice, PKCδ--KO mice are relatively resistant to ethanol intoxication and ethanol is unable to enhance tonic GABA currents in thalamic relay and hippocampal dentate gyrus neurons from PKCδ--KO mice. Mice that are homozygous for this allele are viable and fertile.
A targeting vector was designed to insert a loxP site upstream of exon 2, and a loxP-flanked neomycin resistance (neo) cassette downstream of exon 2 of the protein kinase C, delta (Prkcd) gene. The construct was electroporated into 129X1/SvJ-derived embryonic stem (ES) cells and correctly targeted ES cells were transiently transfected with a pPac-CRE plasmid to remove exon 2 and the neo cassette. Resulting ES cells were injected into blastocysts and resulting chimeric mice were bred with 129X1 mice. PKCδ- mice were crossed to C57BL/6J for three generations and were maintained on a mixed B6;129X1 background. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 1, Robert O Messing|
|Allele Type||Targeted (Null/Knockout)|
|Allele Synonym(s)||PKC delta-; PKCdelta-|
|Gene Symbol and Name||Prkcd, protein kinase C, delta|
|Strain of Origin||129X1/SvJ|
|Molecular Note||Exon 2 was floxed and removed via transient cre-mediated recombination. Exon 2 contains the translation start codon. Northern blot showed absence of transcript in mutants and Western blot confirmed absence of protein in mutants.|
When maintaining a live colony, homozygous mice may be bred together. The donating investigator reported that when maintained on a coisogenic 129X1 background high neonatal mortality and lack of nurturing were observed.
When using the PKCδ-null mouse strain in a publication, please cite the originating article(s) and include JAX stock #028055 in your Materials and Methods section.