Effects of sodium arsenite on the cytoskeleton and cellular glutathione levels in cultured cells.
The effects of As3+ (NaAsO2) on the microtubule and microfilament organization, cytoskeletal protein synthesis, cytoskeletal and cytosolic (soluble) protein sulfhydryls, and cellular glutathione (GSH) levels were examined in Swiss 3T3 mouse cells. Exposure of cells to 2.5 microM As3+ for 16 hr resulted in apparent cell retraction and loss of thick cables of actin filaments. However, the cells still retained numerous thinner microfilaments distributed in a disorganized manner. Microtubule organization was relatively undisturbed. At higher doses (greater than or equal to 20 microM), As3+ treatment caused a severe loss of microtubules and the remaining dense finer actin filaments formed smearing clusters in perinuclear areas. Treatment of cells with As3+ also induced a dose-dependent inhibition of cytoskeletal protein synthesis. Furthermore, As3+ exposure enhanced cellular GSH synthesis since the elevated cellular GSH content in As(3+)-treated cells could be abolished by treatment with buthionine sulfoximine, an inhibitor of gamma-glutamylcysteine synthetase required for GSH biosynthesis. As determined by the N-[3H]-ethylmaleimide binding assay, As3+ exposure also increased the amount of protein sulfhydryls in both the cytoskeletal and the cytosolic protein fractions. Moreover, a greater increase in protein sulfhydryls occurred in the cytoskeletal fraction than in the soluble fraction. These results indicate that the cytoskeleton could be a cellular target for injury by As3+ exposure. The elevated cellular GSH content induced by As3+ could provide a protective mechanism against further injury from this metal insult.
Li
W
W
Department of Microbiology, Boston University School of Medicine, Massachusetts 02118.
Chou
I N
IN
D008841:Actin Cytoskeleton / Q000187:drug effects; D000818:Animals; D001151:Arsenic / Q000494:pharmacology*; D018053:Arsenites; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000096:biosynthesis; D003599:Cytoskeleton / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D005978:Glutathione / Q000378:metabolism*; D051379:Mice; D008868:Microtubule Proteins / Q000096:biosynthesis; D017670:Sodium Compounds; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism
D018053:Arsenites; D003598:Cytoskeletal Proteins; D008868:Microtubule Proteins; D017670:Sodium Compounds; C017947:sodium arsenite; D005978:Glutathione; D001151:Arsenic