The nucleocytoplasmic microfilament network in protoplasts from cultured soybean cells is a plastic entity that pervades the cytoplasm except the central vacuole.
The microfilament network of cultured Glycine max cells (SB-1 line), and protoplasts was visualized with rhodamine-phalloidin under conditions that lysed the protoplast and changed the cell shape. The whole cell had the typical microfilament distribution of a "cage" around the nucleus, from which the large subcortical cables and transvacuolar strands radiated towards the cortex until it reached the cortical microfilament network. Upon cell wall removal, the network conserved its compartmentalization. Thus, the redistribution of the shape where the vacuole becomes a central entity, made the cytoplasm displace peripherally, but the network distribution was conserved. When protoplasts were lysed in a low osmotic medium, the vacuoles were gradually released intact. Under these conditions, the F-actin staining remained within the ghost of the cell, but none was detected in either emerging or almost completely released vacuoles. Most of the released F-actin was found in debris from the cell lysate in the form of microfilaments. When the ghosts were constrained in a coverslip with an air bubble, the shape of the ghost changed accordingly, but the microfilament network distribution remained constant. These results provide further evidence that the vacuole of plant cells does not have detectable associated F-actin. In addition, we demonstrate that the actin microfilament network is a moldable entity that can change its shape but keeps its distribution under constant conditions, in these cultured cells.
Villanueva
Marco A
MA
Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, U.N.A.M., Apartado Postal 510-3, Cuernavaca, Morelos 62250, Mexico. macro@ibt.unam.mx
Schindler
Melvin
M
Wang
John L
JL
D008841:Actin Cytoskeleton / Q000737:chemistry; D000199:Actins / Q000737:chemistry; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D008854:Microscopy, Electron; D010590:Phalloidine / Q000737:chemistry / Q000494:pharmacology; D011485:Protein Binding; D011523:Protoplasts / Q000378:metabolism; D011817:Rabbits; D012235:Rhodamines / Q000737:chemistry / Q000494:pharmacology; D013025:Glycine max / Q000378:metabolism*; D014617:Vacuoles / Q000737:chemistry
D000199:Actins; D005456:Fluorescent Dyes; D012235:Rhodamines; D010590:Phalloidine