A voltage-dependent Ca²⁺ homeostat operates in the plant vacuolar membrane

Cytosolic calcium signals are evoked by a large variety of biotic and abiotic stimuli and play an important role in cellular and long distance signalling in plants. While the function of the plasma membrane in cytosolic Ca²⁺ signalling has been intensively studied, the role of the vacuolar membrane remains elusive. A newly developed vacuolar voltage clamp technique was used in combination with live-cell imaging, to study the role of the vacuolar membrane in Ca²⁺ and pH homeostasis of bulging root hair cells of Arabidopsis. Depolarisation of the vacuolar membrane caused a rapid increase in the Ca²⁺ concentration and alkalised the cytosol, while hyperpolarisation led to the opposite responses. The relationship between the vacuolar membrane potential, the cytosolic pH and Ca²⁺ concentration suggests that a vacuolar H⁺ /Ca²⁺ exchange mechanism plays a central role in cytosolic Ca²⁺ homeostasis. Mathematical modelling further suggests that the voltage-dependent vacuolar Ca²⁺ homeostat could contribute to calcium signalling when coupled to a recently discovered K⁺ channel-dependent module for electrical excitability of the vacuolar membrane.