Current Injection Provokes Rapid Expansion of the Guard Cell Cytosolic Volume and Triggers Ca(2+) Signals

Molecular Plant
Lena J VossM Rob G Roelfsema

Abstract

High-resolution microscopy opens the door for detailed single-cell studies with fluorescent reporter dyes and proteins. We used a confocal spinning disc microscope to monitor fluorescent dyes and the fluorescent protein Venus in tobacco and Arabidopsis guard cells. Multi-barreled microelectrodes were used to inject dyes and apply voltage pulses, which provoke transient rises in the cytosolic Ca(2+) level. Voltage pulses also caused changes in the distribution of Lucifer Yellow and Venus, which pointed to a reversible increase of guard cell cytosolic volume. The dynamic cytosolic volume changes turned out to be provoked by current injection of ions. A reduction of the clamp current, by blocking K(+) uptake channels with Cs(+), strongly suppressed the cytosolic volume changes. Cs(+) not only inhibited the expansion of the cytosol, but also inhibited hyperpolarization-induced elevations of the cytosolic Ca(2+) concentration. A complete loss of voltage-induced Ca(2+) signals occurred when Ca(2+)-permeable plasma membrane channels were simultaneously blocked with La(3+). This shows that two mechanisms cause hyperpolarization-induced elevation of the cytosolic Ca(2+)-concentration: (i) activation of voltage-dependent Ca(2+)-permeable...Continue Reading

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Citations

Jun 26, 2018·The New Phytologist·Dorothea GrausRainer Hedrich
Apr 28, 2018·Journal of Experimental Botany·K R KonradR Hedrich
Feb 13, 2021·The New Phytologist·Julian DindasM Rob G Roelfsema
Jul 1, 2021·Bioscience, Biotechnology, and Biochemistry·Md Tahjib-Ul-ArifYoshiyuki Murata

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