Lipid order in hepatocyte plasma membrane blebs during ATP depletion measured by digitized video fluorescence polarization microscopy

Kathryn Florine-Casteel, John J. Lemasters, Brian Herman

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

Low-light digitized video fluorescence polarization microscopy was used to measure lipid order parameters in plasma membrane blebs of single, cultured rat hepatocytes during ATP depletion with the metabolic inhibitors cyanide and iodoacetic acid. Hepatocytes were labeled on the microscope stage with the plasma membrane probe trimethylammoniumdiphenylhexatriene at successive stages of cell injury. A pair of fluorescence polarization ratio images was obtained from a series of four fluorescence images recorded with a polarizer in the emission path oriented first parallel and then perpendicular to each of two orthogonal excitation light polarization directions. From the polarization ratio images, the lipid order parameter S was determined in individual plasma membrane blebs. Results indicate that the plasma membrane becomes uniformly rigid within a few minutes of the addition of metabolic inhibitors when small surface blebs have formed and ATP levels have fallen by >95%. The measured order parameter of S ~ 0.95 in plasma membrane blebs, compared with S ~ 0.75 in normoxic cell plasma membranes, remained unchanged throughout the course of bleb development and ultimate cell death. These findings demonstrate that significant alteration in hepatocyte plasma membrane structure occurs early in hypoxic cell injury.

Original languageEnglish (US)
Pages (from-to)2078-2084
Number of pages7
JournalFASEB Journal
Volume5
Issue number7
DOIs
StatePublished - 1991

Keywords

  • Digitized video microscopy
  • Fluorescence polarization
  • Hypoxia
  • Membrane fluidity
  • Trimethylammoniumdiphenylhexatriene

Fingerprint Dive into the research topics of 'Lipid order in hepatocyte plasma membrane blebs during ATP depletion measured by digitized video fluorescence polarization microscopy'. Together they form a unique fingerprint.

  • Cite this