Red but not dead? Membranes of stressed Saccharomyces cerevisiae are permeable to propidium iodide

Abstract

Summary: Flow cytometric monitoring of propidium iodide (PI) uptake is a well‐established and rapid method for monitoring cell death and is used on the basis that the intact membrane of viable cells excludes the propidium ion and that loss of this permeability barrier represents irreparable damage and thus cell death. These assumptions are typically based on analysis of live and killed cells. Here we have identified stress levels that lead to a loss of viability of a proportion of Saccharomyces cerevisiae cells and under these conditions we show that there is a subpopulation of cells that can take up PI during and immediately following exposure to stress but that a short incubation allows repair of the membrane damage such that subsequent exposure to PI does not result in staining. Irrespective of the stress applied, approximately 7% of cells exhibited the ability to repair. These results indicate that the level of damage that the yeast cell membrane can sustain and yet retain the ability to repair is greater than previously recognized and care must therefore be taken in using the terms ‘PI‐positive’ and ‘dead’ synonymously. We discuss these findings in the context of the potential for such environmental stress‐induced, transient membrane permeability to have evolutionary implications via the facilitation of horizontal gene transfer.