Okadaic acid induces the rapid and reversible disruption of the neurofilament network in rat dorsal root ganglion neurons.
Treatment of 15-17 day old dissociated cultures of rat dorsal root ganglia with 1 microM okadaic acid caused a reduction in the mobilities of neurofilament subunits on SDS-polyacrylamide gels, signifying an increase in their phosphorylation levels. When cultures were exposed to okadaic acid for 0.5 hrs and harvested in buffer containing Triton X-100, NF-H was nearly completely redistributed to the detergent- soluble fraction while NF-M and NF-L required a longer exposure to the drug before undergoing a similar shift. This redistribution of subunits corresponded with striking changes in the immunofluorescence staining pattern for neurofilaments. Upon removal of okadaic acid from the culture medium following a 0.5 hr treatment, NF-L and NF-M returned to the Triton X-100 insoluble fraction within 2 hrs while NF-H required 10 hrs for recovery.
Sacher
M G
MG
McGill University, Department of Biochemistry, Montreal, Quebec, Canada.
Athlan
E S
ES
Mushynski
W E
WE
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003902:Detergents; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian; D004988:Ethers, Cyclic / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000378:metabolism / Q000648:ultrastructure*; D007382:Intermediate Filaments / Q000187:drug effects / Q000648:ultrastructure*; D016900:Neurofilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D017830:Octoxynol; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011092:Polyethylene Glycols; D051381:Rats
D003902:Detergents; D004988:Ethers, Cyclic; D016900:Neurofilament Proteins; D019319:Okadaic Acid; D011092:Polyethylene Glycols; D017830:Octoxynol; D010749:Phosphoprotein Phosphatases