Axonal localization of delta Ca2+/calmodulin-dependent protein kinase II in developing P19 neurons.

Ca(2+)/calmodulin-dependent protein kinase, type II (CaMK-II) is an enzyme encoded by four genes (alpha, beta, gamma and delta) and traditionally associated with synaptic function in the adult central nervous system, but also believed to play a role during neuronal development. P19 mouse embryonic cells are a model system for neurogenesis and primarily express isozymes of delta CaMK-II. It is not yet known whether or where delta CaMK-II is expressed in P19 neurons. Using an antibody specific for the delta CaMK-II C-terminal tail, we detected a 20-fold increase in levels of delta CaMK-II along axons after 8 days of development. This coincides with increased mRNA and protein levels of delta(C) CaMK-II, which contains the alternative tail. This follows the initial stages of neurite outgrowth and beta(3) tubulin expression, which occur after 4 days. delta CaMK-II co-localizes with the axonal protein GAP-43, but not the dendritic microtubule-associated protein MAP-2, a known substrate of alpha CaMK-II. Like delta CaMK-II, GAP-43 shows increased expression after 8 days. These findings demonstrate developmental regulation of the alternative C-terminal delta CaMK-II exon and implicate endogenous delta CaMK-II in axonal development in embryonic cells.