Calcium/calmodulin-dependent protein kinase kinase β is neuroprotective in stroke in aged mice.

Abstract

Stroke is a devastating neurological disease and the leading cause of long-term disability, particularly in the elderly. Calcium/calmodulin-dependent protein kinase kinase β (CaMKK β) is a major kinase activated by elevated levels of intracellular calcium. Our previous findings in young mice have suggested that CaMKK β is neuroprotective as KO mice had worse stroke outcomes. Because age is an important determinant of stroke outcome, we evaluated the functional role of CaMKK β in stroke in aged mice. We used middle cerebral artery occlusion to induce stroke in aged wild-type (WT) and CaMKK β KO male mice. Lentiviral vectors carrying CaMKK β (LV-CaMKK β) were used to overexpress CaMKK β in the mouse brain. Baseline levels of CaMKK β in the aged brain were significantly lower than those in young mice. LV-CaMKK β treatment reduced infarcts and neurological deficits assessed 3days after stroke. In chronic survival experiments, CaMKK β KO mice showed increased tissue loss in the ipsilateral hemisphere 3weeks after stroke. In addition, KO mice showed poorer functional recovery during the 3-week survival period, as measured by the rotarod test, corner test, locomotor activity assay, and novel object recognition test, compared with WT controls. The loss of blood-brain barrier proteins, inactivation of survival gene expression such as B-cell lymphoma 2 (Bcl-2) and an increase in inflammatory cytokines in the serum were observed after stroke with CaMKK β inhibition. We demonstrate that CaMKK β is neuroprotective in stroke in aged mice. Therefore, our data suggest that CaMKK β may be a potential target for reducing long-term disability after stroke.