Simultaneous detection of CA 2+ and diacylglycerol signaling in living cells

Abstract

Phospholipase C produces two second messengers - diacylglycerol (DAG), which remains in the membrane, and inositol triphosphate (IP 3), which triggers the release of calcium ions (Ca 2+) from intracellular stores. Genetically encoded sensors based on a single circularly permuted fluorescent protein (FP) are robust tools for studying intracellular Ca 2+ dynamics. We have developed a robust sensor for DAG based on a circularly permuted green FP that can be co-imaged with the red fluorescent Ca 2+ sensor R-GECO for simultaneous measurement of both second messengers. © 2012 Tewson et al.