Ld. Johnson et al., delta Ca2+/calmodulin-dependent protein kinase II isozyme-specific induction of neurite outgrowth in P19 embryonal carcinoma cells, J NEUROCHEM, 75(6), 2000, pp. 2380-2391
Ca2+/calmodulin-dependent protein kinase II (CaMK-II) has been linked to th
e induction of differentiation in preneuronal cells. In these cells, delta
isozymes represent the majority of CaMK-IIs expressed and are activated by
differentiation stimuli. To determine whether delta CaMK-IIs are causative
or coincident with in vitro differentiation, we overexpressed wild-type, co
nstitutively active, and C-terminal domains of delta and gamma CaMK-II isoz
ymes in mouse P19 and NIH/3T3 cells using high-efficiency transfections. At
1-2 days after transfection, only constitutively active delta CaMK-II isoz
ymes induced branched cellular extensions in both cell types. In P19 cells,
retinoic acid induced neurite extensions after 3-4 days; these extensions
were coincident with a fourfold increase in endogenous CaMK-II activity. Ex
tensions induced by both retinoic acid and delta CaMK-IIs contained class I
II beta -tubulin in a discontinuous or beaded pattern. C-terminal CaMK-II c
onstructs disrupted the ability of endogenous CaMK-II to autophosphorylate
and blocked retinoic acid-induced differentiation. delta CaMK-II was found
along extensions, whereas gamma CaMK-II exhibited a more diffuse, cytosolic
localization. These data not only support an extranuclear role for CaMK-II
in promoting neurite outgrowth, but also demonstrate CaMK-II isozyme speci
ficity in these early steps of neuronal differentiation.