Calcium imaging in Arabidopsis pollen cells using G-CaMP5.

Abstract

Calcium (Ca2+ ) signaling has been implicated in pollen germination and pollen tube growth. To date, however, we still know very little about how exactly Ca2+ signaling links to various physiological subcellular processes during pollen germination and pollen tube growth. Given that Ca2+ signaling is tightly related to the cytosolic concentration and dynamics of Ca2+ , it is vital to trace the dynamic changes in Ca2+ levels in order to decode Ca2+ signaling. Here, we demonstrate that G-CaMP5 serves well as an indicator for monitoring cytosolic Ca2+ dynamics in pollen cells. Using this probe, we show that cytosolic Ca2+ changes dramatically during pollen germination, and, as reported previously, Ca2+ forms a tip-focused gradient in the pollen tube and undergoes oscillation in the tip region during pollen tube growth. In particular, using G-CaMP5 allowed us to capture the dynamic changes in the cytosolic Ca2+ concentration ([Ca2+ ]cyt ) in pollen tubes in response to various exogenous treatments. Our data suggest that G-CaMP5 is a suitable probe for monitoring the dynamics of [Ca2+ ]cyt in pollen cells.