期刊
FEBS OPEN BIO
卷 4, 期 -, 页码 190-199出版社
WILEY
DOI: 10.1016/j.fob.2014.02.003
关键词
Cell fixation; Cell blebbing; Phosphatidylinositol 4,5-bisphosphate (PIP2); Human umbilical vein endothelial cells (HUVECs); Lipid rafts; THP-1-derived macrophages
资金
- National Natural Science Foundation of China [31260205]
- Scientific Research Foundation for Returned Overseas Chinese Scholar of State Education Ministry
While most attention has been focused on physiologically generated blebs, the molecular mechanisms for fixation-induced cell blebbing are less investigated. We show that protein-fixing (e.g. aldehydes and picric acid) but not lipid-stabilizing (e.g. OsO4 and KMnO4) fixatives induce blebbing on spread cells. We also show that aldehyde fixation may induce the loss or delocalization of phosphatidylinositol 4,5-bisphosphate (PIP2) in the plasma membrane and that the asymmetric distribution of fixation-induced blebs on spread/ migrating cells coincides with that of PIP2 on the cells prefixed by lipid-stabilizing fixatives (e.g., OsO4). Moreover, fixation induces blebbing less readily on PIP2-elevated spread cells but more readily on PIP2-lowered or lipid raft-disrupted spread cells. Our data suggest that fixation-induced lowering of PIP2 level at cytoskeleton-attaching membrane sites causes bleb formation via local breakdown of the membrane-cytoskeleton coupling. (C) 2014 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
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