Neutrophil protein kinase Cδ as a mediator of stroke-reperfusion injury
Wen-Hai Chou, … , Donna M. Ferriero, Robert O. Messing
Wen-Hai Chou, … , Donna M. Ferriero, Robert O. Messing
Published July 1, 2004
Citation Information: J Clin Invest. 2004;114(1):49-56. https://doi.org/10.1172/JCI21655.
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Article Neuroscience

Neutrophil protein kinase Cδ as a mediator of stroke-reperfusion injury

Abstract

Thrombolysis is widely used to intervene in acute ischemic stroke, but reestablishment of circulation may paradoxically initiate a reperfusion injury. Here we describe studies with mice lacking protein kinase Cδ (PKCδ) showing that absence of this enzyme markedly reduces reperfusion injury following transient ischemia. This was associated with reduced infiltration of peripheral blood neutrophils into infarcted tissue and with impaired neutrophil adhesion, migration, respiratory burst, and degranulation in vitro. Total body irradiation followed by transplantation with bone marrow from PKCδ-null mice donors reduced infarct size and improved neurological outcome in WT mice, whereas marrow transplantation from WT donors increased infarction and worsened neurological scores in PKCδ-null mice. These results indicate an important role for neutrophil PKCδ in reperfusion injury and strongly suggest that PKCδ inhibitors could prove useful in the treatment of stroke.

Authors

Wen-Hai Chou, Doo-Sup Choi, Hong Zhang, Dezhi Mu, Tom McMahon, Viktor N. Kharazia, Clifford A. Lowell, Donna M. Ferriero, Robert O. Messing

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Figure 1

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Generation of PKCδ-null mice. (A) Organization of the mouse PKCδ gene, t...
Generation of PKCδ-null mice. (A) Organization of the mouse PKCδ gene, the targeting construct, and the allele resulting from homologous recombination. Boxes represent exon 1 (123 bp) and exon 2 (200 bp). Arrowheads represent loxP sequences. Only relevant restriction enzyme sites are indicated. E, EcoRI; H, HindIII; B, BamHI; S, SacI; N, NotI. (B) Verification of genotypes by PCR using the primers P7 and P3. The WT allele (+/+) generates a 2.9-kb and the mutant allele (−/−) a 1.8-kb band, as indicated. (C) Verification of genotypes by Southern blot analysis of genomic DNA digested with EcoRI from WT (+/+), heterozygous (+/ –), and homozygous mutants ( –/ –) using a 3.2-kb 5′-probe (HindIII-SacI fragment). (D) Verification of genotypes by Western blot analysis of whole brain lysates using a mAb against PKCδ. The migration of PKCδ immunoreactivity (78 kDa) is indicated.