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Fixation‐induced cell blebbing on spread cells inversely correlates with phosphatidylinositol 4,5‐bisphosphate level in the plasma membrane

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. OsO and KMnO) fixatives induce blebbing on spread... Full description

Journal Title: FEBS Open Bio 01 January 2014, Vol.4(1), pp.190-199
Main Author: Zhao, Siyuan
Other Authors: Liao, Huanhuan , Ao, Meiying , Wu, Li , Zhang, Xiaojun , Chen, Yong
Format: Electronic Article Electronic Article
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ID: ISSN: 2211-5463 ; E-ISSN: 2211-5463 ; DOI: 10.1016/j.fob.2014.02.003
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recordid: wj10.1016/j.fob.2014.02.003
title: Fixation‐induced cell blebbing on spread cells inversely correlates with phosphatidylinositol 4,5‐bisphosphate level in the plasma membrane
format: Article
creator:
  • Zhao, Siyuan
  • Liao, Huanhuan
  • Ao, Meiying
  • Wu, Li
  • Zhang, Xiaojun
  • Chen, Yong
subjects:
  • Cell Fixation
  • Cell Blebbing
  • Phosphatidylinositol 4
  • 5-Bisphosphate Pip 2
  • Human Umbilical Vein Endothelial Cells Huvecs
  • Lipid Rafts
  • Thp-1-Derived Macrophages
ispartof: FEBS Open Bio, 01 January 2014, Vol.4(1), pp.190-199
description: 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. OsO and KMnO) fixatives induce blebbing on spread cells. We also show that aldehyde fixation may induce the loss or delocalization of phosphatidylinositol 4,5‐bisphosphate (PIP) in the plasma membrane and that the asymmetric distribution of fixation‐induced blebs on spread/migrating cells coincides with that of PIP on the cells prefixed by lipid‐stabilizing fixatives (e.g., OsO). Moreover, fixation induces blebbing less readily on PIP‐elevated spread cells but more readily on PIP‐lowered or lipid raft‐disrupted spread cells. Our data suggest that fixation‐induced lowering of PIP level at cytoskeleton‐attaching membrane sites causes bleb formation via local breakdown of the membrane–cytoskeleton coupling. Protein‐ but not lipid‐stabilizing fixatives induce cell blebbing of spread cells. Asymmetric distribution of fixation‐induced blebs coincides with that of PIP2. Fixation less readily induces blebbing on spread cells with elevated PIP2 levels. Fixation more readily induces blebbing on spread cells with lower PIP2 levels. Disruption of lipid rafts enhances fixation‐induced blebbing of spread cells.
language:
source:
identifier: ISSN: 2211-5463 ; E-ISSN: 2211-5463 ; DOI: 10.1016/j.fob.2014.02.003
fulltext: fulltext
issn:
  • 2211-5463
  • 22115463
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titleFixation‐induced cell blebbing on spread cells inversely correlates with phosphatidylinositol 4,5‐bisphosphate level in the plasma membrane
creatorZhao, Siyuan ; Liao, Huanhuan ; Ao, Meiying ; Wu, Li ; Zhang, Xiaojun ; Chen, Yong
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subjectCell Fixation ; Cell Blebbing ; Phosphatidylinositol 4 ; 5-Bisphosphate Pip 2 ; Human Umbilical Vein Endothelial Cells Huvecs ; Lipid Rafts ; Thp-1-Derived Macrophages
descriptionWhile 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. OsO and KMnO) fixatives induce blebbing on spread cells. We also show that aldehyde fixation may induce the loss or delocalization of phosphatidylinositol 4,5‐bisphosphate (PIP) in the plasma membrane and that the asymmetric distribution of fixation‐induced blebs on spread/migrating cells coincides with that of PIP on the cells prefixed by lipid‐stabilizing fixatives (e.g., OsO). Moreover, fixation induces blebbing less readily on PIP‐elevated spread cells but more readily on PIP‐lowered or lipid raft‐disrupted spread cells. Our data suggest that fixation‐induced lowering of PIP level at cytoskeleton‐attaching membrane sites causes bleb formation via local breakdown of the membrane–cytoskeleton coupling. Protein‐ but not lipid‐stabilizing fixatives induce cell blebbing of spread cells. Asymmetric distribution of fixation‐induced blebs coincides with that of PIP2. Fixation less readily induces blebbing on spread cells with elevated PIP2 levels. Fixation more readily induces blebbing on spread cells with lower PIP2 levels. Disruption of lipid rafts enhances fixation‐induced blebbing of spread cells.
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descriptionWhile 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. OsO and KMnO) fixatives induce blebbing on spread cells. We also show that aldehyde fixation may induce the loss or delocalization of phosphatidylinositol 4,5‐bisphosphate (PIP) in the plasma membrane and that the asymmetric distribution of fixation‐induced blebs on spread/migrating cells coincides with that of PIP on the cells prefixed by lipid‐stabilizing fixatives (e.g., OsO). Moreover, fixation induces blebbing less readily on PIP‐elevated spread cells but more readily on PIP‐lowered or lipid raft‐disrupted spread cells. Our data suggest that fixation‐induced lowering of PIP level at cytoskeleton‐attaching membrane sites causes bleb formation via local breakdown of the membrane–cytoskeleton coupling. Protein‐ but not lipid‐stabilizing fixatives induce cell blebbing of spread cells. Asymmetric distribution of fixation‐induced blebs coincides with that of PIP2. Fixation less readily induces blebbing on spread cells with elevated PIP2 levels. Fixation more readily induces blebbing on spread cells with lower PIP2 levels. Disruption of lipid rafts enhances fixation‐induced blebbing of spread cells.
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abstractWhile 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. OsO and KMnO) fixatives induce blebbing on spread cells. We also show that aldehyde fixation may induce the loss or delocalization of phosphatidylinositol 4,5‐bisphosphate (PIP) in the plasma membrane and that the asymmetric distribution of fixation‐induced blebs on spread/migrating cells coincides with that of PIP on the cells prefixed by lipid‐stabilizing fixatives (e.g., OsO). Moreover, fixation induces blebbing less readily on PIP‐elevated spread cells but more readily on PIP‐lowered or lipid raft‐disrupted spread cells. Our data suggest that fixation‐induced lowering of PIP level at cytoskeleton‐attaching membrane sites causes bleb formation via local breakdown of the membrane–cytoskeleton coupling. Protein‐ but not lipid‐stabilizing fixatives induce cell blebbing of spread cells. Asymmetric distribution of fixation‐induced blebs coincides with that of PIP2. Fixation less readily induces blebbing on spread cells with elevated PIP2 levels. Fixation more readily induces blebbing on spread cells with lower PIP2 levels. Disruption of lipid rafts enhances fixation‐induced blebbing of spread cells.
doi10.1016/j.fob.2014.02.003
pages190-199
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date2014-01-01