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Plasma membrane damage detected by nucleic acid leakage
1Department of Biology, University of Padua, Padova, Italy.
Molecular Toxicology
|January 1, 1989
Summary
This study introduces a novel protocol to detect and quantify cell membrane damage by measuring the release of intracellular components. The method distinguishes between mild and severe membrane damage, differentiating permeabilization from cell lysis.
Area of Science:
- Cell Biology
- Toxicology
- Biochemistry
Background:
- Cell membrane integrity is crucial for cellular function.
- Membrane damage can be assessed by the release of intracellular components.
- Quantifying released components helps evaluate the extent and type of membrane damage.
Purpose of the Study:
- To develop and validate a protocol for detecting and quantifying cell membrane damage.
- To differentiate between generalized membrane permeabilization and cell lysis.
- To assess the impact of various chemicals on cell membrane integrity.
Main Methods:
- Cells were preincubated with tritiated adenine, followed by a chase in nonradioactive medium.
- Radioactivity in released components (medium, nucleotide pool, RNA, DNA) was measured.
- Analysis of macromolecular content in released material and cell-associated radioactivity quantified damage.
Main Results:
- The protocol effectively measures radioactivity released into the medium, reflecting membrane damage.
- Analysis of released macromolecules distinguishes mild from severe membrane damage.
- Variations in RNA and DNA radioactivity differentiate cell permeabilization from lysis.
Conclusions:
- The developed protocol provides a quantitative method to assess cell membrane damage.
- It allows for the distinction between different mechanisms of membrane damage, such as permeabilization versus lysis.
- The protocol is applicable for evaluating the effects of various toxicants, including detergents and Cr(VI).