title: CaMKII activation is a novel effector of alcohol's neurotoxicity in neural crest stem/progenitor cells.
abstract: Prenatal ethanol exposure causes significant neurodevelopmental deficits through its induction of apoptosis in neuronal progenitors including the neural crest. Using an established chick embryo model, we previously showed that clinically relevant ethanol concentrations cause neural crest apoptosis through mobilization of an intracellular calcium transient. How the calcium transient initiates this cell death is unknown. In this study, we identify CaMKII as the calcium target responsible for ethanol-induced apoptosis. Immunostaining revealed selective enrichment of activated phosphoCaMKII(Thr286) within ethanol-treated neural crest. CaMKII activation in response to ethanol was rapid (< 60 s) and robust, and CaMKII activity was increased 300% over control levels. Treatment with CaMKII-selective inhibitors but not those directed against CaMKIV or PKC completely prevented the cell death. Forced expression of dominant-negative CaMKII prevented ethanol's activation of CaMKII and prevented the ethanol-induced death, whereas constitutively active CaMKII in ethanol's absence significantly increased cell death to levels caused by ethanol treatment. In summary, CaMKII is the key signal that converts the ethanol-induced, short-lived Ca(i) (2+) transient into a long-lived cellular effector. This is the first identification of CaMKII as a critical mediator of ethanol-induced cell death. Because neural crest differentiates into several neuronal lineages, our findings offer novel insights into how ethanol disrupts early neurogenesis.
authors.affiliation: Department of Nutritional Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
authors.forename: Ana
authors.initials: A
authors.lastname: Garic
authors.forename: George R
authors.initials: GR
authors.lastname: Flentke
authors.forename: Ed
authors.initials: E
authors.lastname: Amberger
authors.forename: Marcos
authors.initials: M
authors.lastname: Hernandez
authors.forename: Susan M
authors.initials: SM
authors.lastname: Smith
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism*; D002492:Central Nervous System Depressants / Q000633:toxicity*; D002642:Chick Embryo; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects*; D000431:Ethanol / Q000633:toxicity*; D007150:Immunohistochemistry; D009432:Neural Crest / Q000187:drug effects* / Q000473:pathology*; D058953:Neural Stem Cells / Q000187:drug effects* / Q000473:pathology*; D011493:Protein Kinase C / Q000378:metabolism; D013268:Stimulation, Chemical
chemical_list: D002492:Central Nervous System Depressants; D018076:DNA, Complementary; D000431:Ethanol; D011493:Protein Kinase C; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D002118:Calcium