In vitro calcium and calmodulin-dependent kinase-mediated phosphorylation of rat brain and spinal cord neurofilament proteins is increased by glycidamide administration.
This study was carried out to determine the action of glycidamide (2,3-epoxy-1-propanamide), a neurotoxic metabolite of acrylamide, on Ca2+/calmodulin (CaM)-dependent protein kinase phosphorylation of cytoskeletal proteins. Acrylamide has been shown to increase Ca2+/CaM-dependent phosphorylation of neurofilament (NF) triplet proteins and autophosphorylation of Ca2+/CaM-dependent protein kinase II (CaM kinase II; EC 2.7.1.37). A daily intraperitoneal dose of 0.7 mmol/kg b.wt. of glycidamide or deionized water was administered to male Sprague-Dawley rats. Animals were sacrificed when signs of severe neurotoxicity became apparent at 13-16 days of treatment. Axonal floatation was used to isolate neurofilaments (NFs) and endogenous kinases from brains and spinal cords of treated and control animals. Samples isolated from brain and spinal cord of glycidamide-treated animals showed increased in vitro Ca2+/CaM-dependent phosphorylation of endogenous and exogenous NF proteins and increased autophosphorylation of CaM kinase II when compared with controls. CaM binding to the alpha, beta, and beta' subunits of CaM kinase II and antibody binding to the alpha-subunit of CaM kinase II in brain supernatant isolates was increased as a result of glycidamide treatment. These results suggest that increased Ca2+/CaM-dependent phosphorylation of cytoskeletal proteins may be involved in the pathogenesis of glycidamide-induced neurotoxicity.
Reagan
K E
KE
Department of Pharmacology, Duke University Medical Center, Durham, NC 27710, USA.
Wilmarth
K R
KR
Friedman
M A
MA
Abou-Donia
M B
MB
D020106:Acrylamide; D000178:Acrylamides / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000201:enzymology / Q000378:metabolism*; D001923:Brain Chemistry; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000494:pharmacology*; D004852:Epoxy Compounds / Q000494:pharmacology*; D007118:Immunoassay; D066298:In Vitro Techniques; D008297:Male; D016900:Neurofilament Proteins / Q000187:drug effects / Q000378:metabolism*; D009498:Neurotoxins / Q000494:pharmacology; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000737:chemistry* / Q000201:enzymology
D000178:Acrylamides; D004852:Epoxy Compounds; D016900:Neurofilament Proteins; D009498:Neurotoxins; D020106:Acrylamide; C071834:glycidamide; D017871:Calcium-Calmodulin-Dependent Protein Kinases