Reactive oxygen species regulate F-actin dynamics in neuronal growth cones and neurite outgrowth.
Reactive oxygen species are well known for their damaging effects due to oxidation of lipids, proteins and DNA that ultimately result in cell death. Accumulating evidence indicates that reactive oxygen species also have important signaling functions in cell proliferation, differentiation, cell motility and apoptosis. Here, we tested the hypothesis whether reactive oxygen species play a physiological role in regulating F-actin structure and dynamics in neuronal growth cones. Lowering cytoplasmic levels of reactive oxygen species with a free radical scavenger, N-tert-butyl-alpha-phenylnitrone, or by inhibiting specific sources of reactive oxygen species, such as NADPH oxidases or lipoxygenases, reduced the F-actin content in the peripheral domain of growth cones. Fluorescent speckle microscopy revealed that these treatments caused actin assembly inhibition, reduced retrograde actin flow and increased contractility of actin structures in the transition zone referred to as arcs, possibly by activating the Rho pathway. Reduced levels of reactive oxygen species ultimately resulted in disassembly of the actin cytoskeleton. When neurons were cultured overnight in conditions of reduced free radicals, growth cone formation and neurite outgrowth were severely impaired. Therefore, we conclude that physiological levels of reactive oxygen species are critical for maintaining a dynamic F-actin cytoskeleton and controlling neurite outgrowth.
Munnamalai
Vidhya
V
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-2054, USA.
Suter
Daniel M
DM
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001048:Aplysia; D002478:Cells, Cultured; D020439:Growth Cones / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents; D008084:Lipoxygenase / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D019255:NADPH Oxidases / Q000378:metabolism; D016501:Neurites / Q000378:metabolism* / Q000648:ultrastructure; D017382:Reactive Oxygen Species / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism
D000199:Actins; D007202:Indicators and Reagents; D017382:Reactive Oxygen Species; D008084:Lipoxygenase; D019255:NADPH Oxidases; D054460:rho-Associated Kinases