Genetically encoded voltage indicator revealed differential effects of hyperosmotic stress on yeast plasma membrane potential imposed by different stress conditions

Abstract

Abstract Cells can be affected by several causes of osmotic stress by which they were forced to tolerate and adapt. An essential aspect of adaptation is ion regulation, and many insights into such complex processes can be obtained through measurement of plasma membrane potential (PMP) of cells during the stress. We recently established genetically encoded voltage indicator proteins which could be utilized to report yeast PMP change in real time. In this work, we employed this method to monitor early change in PMP of yeast Saccharomyces cerevisiae with intact cell wall, immediately following hyperosmotic up-shock due to various stress agents. The results pointed to differential effects of NaCl, sorbitol, and Polyethylene Glycol (PEG) 6000. Yeast PMP was more responsive toward PEG 6000 than NaCl and sorbitol at comparable osmotic pressure, and PEG 6000 stimulated the largest response magnitude, followed by sorbitol and NaCl, respectively. After prolonged treatments, PEG 6000 also instigated distinct cell morphology from NaCl and sorbitol. Accordingly, this study presents new evidence supporting multiple pathways underlying yeast adaptation to varying hyperosmotic conditions, enabled through the optical physiology approach. Our findings promote better understanding of yeast cellular response to hyperosmotic stress, with tenable relevance to the physiologically-related plant cells.