Phosphorylation of calcium calmodulin-dependent protein kinase II following lateral fluid percussion brain injury in rats.
Traumatic brain injury (TBI) can dramatically increase levels of intracellular calcium ([Ca(2+)](i)). One consequence of increased [Ca(2+)](i) would be altered activity and function of calcium-regulated proteins, including calcium-calmodulin-dependent protein kinase II (CaMKII), which is autophosphorylated on Thr(286)(pCaMKII(286)) in the presence of calcium and calmodulin. Therefore, we hypothesized that TBI would result in increased levels of pCaMKII(286), and that such increases would occur early after injury in brain regions known to be damaged following lateral fluid percussion TBI (i.e., hippocampus and cortex). In order to test this hypothesis, immunostaining of CaMKII was examined in rat hippocampus and cortex after lateral fluid percussion (LFP) injury using an antibody directed against pCaMKII(286). LFP injury produced a marked increase in pCaMKII(286) immunostaining in the hippocampus and overlying cortex 30 min after TBI. The pattern of increased immunostaining was uneven, and unexpectedly absent in some hippocampal CA3 pyramidal neurons. This suggests that phosphatase activity may also increase following TBI, resulting in dephosphorylation of pCaMKII(286) in subpopulations of CA3 pyramidal neurons. Western blotting confirmed a rapid increase in levels of pCaMKII(286) at 10 and 30 min after brain injury, and that it was transient and no longer significantly elevated when examined at 3, 8, and 24 h. These results demonstrate that TBI alters the autophosphorylation state of CaMKII, an important neuronal regulator of critical cell functions, including enzyme activities, cell structure, gene expression, and neuronal plasticity, and provide a molecular mechanism that is likely to contribute to cell injury and impaired plasticity after TBI.
Folkerts
Michael M
MM
Department of Neurological Surgery, University of California Davis, Davis, California 95616, USA.
Parks
Elizabeth A
EA
Dedman
John R
JR
Kaetzel
Marcia A
MA
Lyeth
Bruce G
BG
Berman
Robert F
RF
D000818:Animals; D048588:Benzoxazines; D015153:Blotting, Western; D001831:Body Temperature / Q000502:physiology; D001835:Body Weight / Q000502:physiology; D001930:Brain Injuries / Q000378:metabolism* / Q000473:pathology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D007839:Functional Laterality / Q000502:physiology; D007150:Immunohistochemistry; D008297:Male; D010078:Oxazines; D010766:Phosphorylation; D004856:Postural Balance / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
D048588:Benzoxazines; D010078:Oxazines; C028911:cresyl violet; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases