A method for analysis of the detergent-resistant cytoskeleton of cells within organs.
We describe a method for the preparation of the detergent-resistant cytoskeleton and nuclear matrix of cells within organs and tissues. Such cells were previously inaccessible to study because the three-dimensional organization of cells in organs prevented uniform distribution of the detergent throughout the multiple cell layers. We use the method presented here to compare the proteins present in the cytoskeleton, nuclear matrix and soluble fractions of cells from different histotypes. SDS-gel analysis demonstrates that soluble and nuclear matrix proteins differ greatly between histotypes while cytoskeletal proteins are relatively similar. Immunocytochemical analysis of tissue prepared using this procedure also demonstrates that the intracellular structure of cells within organs differs from that of in vitro cultured cells.
Capco
D G
DG
Department of Zoology, Arizona State University, Tempe 85287.
Munoz
D M
DM
Gassmann
C J
CJ
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000032:analysis; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron
D003598:Cytoskeletal Proteins; D003902:Detergents