Lippincott Williams & Wilkins, Inc., Philadelphia
δ Ca2+/Calmodulin-Dependent Protein Kinase IIIsozyme-Specific Induction of Neurite Outgrowth in P19 Embryonal Carcinoma Cells
Article first published online: 29 JUL 2008
Journal of Neurochemistry
Volume 75, Issue 6, pages 2380–2391, December 2000
How to Cite
Johnson, L. D., Willoughby, C. A., Burke, S. H., Paik, D. S., Jenkins, K. J. and Tombes, R. M. (2000), δ Ca2+/Calmodulin-Dependent Protein Kinase IIIsozyme-Specific Induction of Neurite Outgrowth in P19 Embryonal Carcinoma Cells. Journal of Neurochemistry, 75: 2380–2391. doi: 10.1046/j.1471-4159.2000.0752380.x
Abbreviations used: ATRA, all-trans-retinoic acid; CaM,calmodulin; CaMK-II, Ca2+/calmodulin-dependent protein kinase II;EGFP, enhanced green fluorescent protein; GFP, green fluorescent protein;TBST, Tris-buffered saline (pH 7.4) and 0.1% Tween 20; βIII tubulin,class III β-tubulin.
- Issue published online: 29 JUL 2008
- Article first published online: 29 JUL 2008
- Ca2+/calmodulin-dependent protein kinase II;
- P19 embryonal carcinoma cells;
- Retinoic acid;
- Green fluorescent protein
Abstract: Ca2+/calmodulin-dependent protein kinase II(CaMK-II) has been linked to the induction of differentiation in preneuronalcells. In these cells, δ isozymes represent the majority of CaMK-IIsexpressed and are activated by differentiation stimuli. To determine whetherδ CaMK-IIs are causative or coincident with in vitro differentiation, weoverexpressed wild-type, constitutively active, and C-terminal domains ofδ and γ CaMK-II isozymes in mouse P19 and NIH/3T3 cells usinghigh-efficiency transfections. At 1-2 days after transfection, onlyconstitutively active δ CaMK-II isozymes induced branched cellularextensions in both cell types. In P19 cells, retinoic acid induced neuriteextensions after 3-4 days; these extensions were coincident with a fourfoldincrease in endogenous CaMK-II activity. Extensions induced by both retinoicacid and δ CaMK-IIs contained class III β-tubulin in adiscontinuous or beaded pattern. C-terminal CaMK-II constructs disrupted theability of endogenous CaMK-II to autophosphorylate and blocked retinoicacid-induced differentiation. δ CaMK-II was found along extensions,whereas γ CaMK-II exhibited a more diffuse, cytosolic localization.These data not only support an extranuclear role for CaMK-II in promotingneurite outgrowth, but also demonstrate CaMK-II isozyme specificity in theseearly steps of neuronal differentiation.