Formation of actin-ADF/cofilin rods transiently retards decline of mitochondrial potential and ATP in stressed neurons.
When neurons in culture are transiently stressed by inhibition of ATP synthesis, they rapidly form within their neurites rodlike actin inclusions that disappear when the insult is removed. Oxidative stress, excitotoxic insults, and amyloid beta-peptide oligomers also induce rods. Immunostaining of neurites indicates that these rods also contain the majority of the actin filament dynamizing proteins, actin-depolymerizing factor (ADF) and cofilin (AC). If the rods reappear within 24 h after the stress is removed, the neurite degenerates distal to the rod but with no increase in neuronal death. Here, rods were generated in cultured rat E18 hippocampal cells by overexpression of a green fluorescent protein chimera of AC. Surprisingly, we have found that, for a short period (approximately 60 min) immediately after initial rod formation, the loss of mitochondrial membrane potential (Delta Psi(m)) and ATP in neurites with rods is slower than in neurites without them. The Delta Psi(m) was monitored with the fluorescent dye tetramethylrhodamine methyl ester, and ATP was monitored with the fluorescent ion indicator mag-fura 2. Actin in rods is less dynamic than is filamentous actin in other cytoskeletal structures. Because Delta Psi(m) depends on cellular ATP and because ATP hydrolysis associated with actin filament turnover is responsible for a large fraction of neuronal energy consumption (approximately 50%), the formation of rods transiently protects neurites by slowing filament turnover and its associated ATP hydrolysis.
Bernstein
Barbara W
BW
Dept. of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523-1870, USA. bwb@lamar.colostate.edu
Chen
Hui
H
Boyle
Judith A
JA
Bamburg
James R
JR
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002478:Cells, Cultured; D051345:Destrin / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology*; D051336:Mitochondrial Membranes / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D021381:Protein Transport / Q000187:drug effects; D011554:Pseudopodia / Q000187:drug effects; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D053778:Thiazolidines / Q000494:pharmacology; D014982:Xenopus laevis
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D051345:Destrin; D011993:Recombinant Fusion Proteins; D053778:Thiazolidines; D000255:Adenosine Triphosphate; C037067:latrunculin A