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Calcium/Calmodulin-Dependent Kinase (CaMKII) Inhibition Protects Against Purkinje Cell Damage Following CA/CPR in Mice

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Abstract

Ischemic brain damage is triggered by glutamate excitotoxicity resulting in neuronal cell death. Previous research has demonstrated that N-methly-d-aspartate (NMDA) receptor activation triggers downstream calcium-dependent signaling pathways, specifically Ca2+/calmodulin-dependent protein kinase II (CaMKII). Inhibiting CaMKII is protective against hippocampal ischemic injury, but there is little known about its role in the cerebellum. To examine the neuroprotective potential of CaMKII inhibition in Purkinje cells, we subjected C57BL/6 or CaMKIIα KO male mice (8–12 weeks old) to cardiac arrest followed by cardiopulmonary resuscitation (CA/CPR). We performed a dose-response study for tat-CN19o and cerebellar injury was analyzed at 7 days after CA/CPR. Acute signaling was assessed at 6 h after CA/CPR using Western blot analysis. We observed increased phosphorylation of the T286 residue of CaMKII, suggesting increased autonomous activation. Analysis of Purkinje cell density revealed a decrease in cell density at 7 days after CA/CPR that was prevented with tat-CN19o at doses of 0.1 and 1 mg/kg. However, neuroprotection in the cerebellum required doses that were 10-fold higher than what was needed in the hippocampus. CaMKIIα KO mice subjected to sham surgery or CA/CPR had similar Purkinje cell densities, suggesting CaMKIIα is required for CA/CPR-induced injury in the cerebellum. We also observed a CA/CPR-induced activation of death-associated protein kinase (DAPK1) that tat-CN19o did not block. In summary, our findings indicate that inhibition of autonomous CaMKII activity is a promising therapeutic approach that is effective across multiple brain regions.

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Funding

This work was supported by NINDS K01 NS086969 (NQ), NINDS R01 NS080851 (PSH, UB) and Gates Summer Internship Program (MR).

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Authors and Affiliations

  1. Neuronal Injury Program, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA

    Nicholas E. Chalmers, Joan Yonchek, Kathryn E. Steklac, Matthew Ramsey, Paco S. Herson & Nidia Quillinan

  2. Department of Anesthesiology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA

    Nicholas E. Chalmers, Joan Yonchek, Paco S. Herson & Nidia Quillinan

  3. Department of Pharmacology, University of Colorado, Anschutz Medical Campus, Aurora, CO, USA

    K. Ulrich Bayer & Paco S. Herson

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  1. Nicholas E. Chalmers
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Correspondence to Nidia Quillinan.

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All experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Colorado, School of Medicine, and were performed according to the guidelines from the National Institutes of Health.

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Chalmers, N.E., Yonchek, J., Steklac, K.E. et al. Calcium/Calmodulin-Dependent Kinase (CaMKII) Inhibition Protects Against Purkinje Cell Damage Following CA/CPR in Mice. Mol Neurobiol 57, 150–158 (2020). https://doi.org/10.1007/s12035-019-01765-9

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