Multiple methods of visualizing the yeast vacuole permit evaluation of its morphology and inheritance during the cell cycle

Abstract

The vacuole of the yeast Saccharomyces cerevisiae was visualized with three unrelated fluorescent dyes: FITC-dextran, quinacrine, and an endogenous fluorophore produced in ade2 yeast. FITC-dextran, which enters cells by endocytosis, had been previously developed as a vital stain for yeast vacuoles. Quinacrine, which diffuses across membranes and accumulates in acidic compartments in mammalian cells, can also be used as a marker for yeast vacuoles. ade2 yeast accumulate an endogenous fluorophore in their vacuoles. Using these stains, yeast were examined for vacuole morphology throughout the cell division cycle. In both the parent cell and the bud, a single vacuole was the most common morphology at every stage. Two or more vacuoles could also be found in the mother cell or in the bud; however, this morphology was not correlated with any stage of the cell division cycle. Even small buds (in early S phase) often contained a small vacuole. By the time the bud was half the diameter of the mother cell, it almost always bore a vacuole. This picture of vacuole division and segregation differs from what is seen with synchronized cultures. In ade2 yeast, the bud usually inherits a substantial portion of its vacuole contents from the mother cell. We propose that vacuolar segregation is accomplished by vesicular traffic between the parent cell and the bud.