Calcium ions (Ca2+) play a key role in cell signaling across organisms. In plants, a plethora of environmental and developmental stimuli induce specific Ca2+ increases in the cytosol as well as in different cellular compartments including the endoplasmic reticulum (ER). The ER represents an intracellular Ca2+ store that actively accumulates Ca2+ taken up from the cytosol. By exploiting state-of-the-art genetically encoded Ca2+ indicators (GECIs), specifically the ER-GCaMP6-210 and R-GECO1, we report the generation and characterization of an Arabidopsis (Arabidopsis thaliana) line that allows for simultaneous imaging of Ca2+ dynamics in both the ER and cytosol at different spatial scales. By performing analyses in single cells, we precisely quantified i) the time required by the ER to import Ca2+ from the cytosol into the lumen; and ii) the time required to observe a cytosolic Ca2+ increase upon the pharmacological inhibition of the ER-localized type IIA Ca2+-ATPases. Furthermore, live imaging of mature, soil-grown plants revealed the existence of a wounding-induced, long-distance ER Ca2+ wave propagating in injured and systemic rosette leaves. This technology enhances high-resolution analyses of intracellular Ca2+ dynamics at the cellular level and in adult organisms and paves the way to develop new methodologies aimed at defining the contribution of subcellular compartments in Ca2+ homeostasis and signaling.

Simultaneous imaging of {ER} and cytosolic Ca2$mathplus$ dynamics reveals long-distance {ER} Ca2$mathplus$ waves in plants / F. Resentini, M. Grenzi, D. Ancora, M. Cademartori, L. Luoni, M. Franco, A. Bassi, M.C. Bonza, A. Costa. - In: PLANT PHYSIOLOGY. - ISSN 1532-2548. - (2021 Jan). [Epub ahead of print]

Simultaneous imaging of {ER} and cytosolic Ca2$mathplus$ dynamics reveals long-distance {ER} Ca2$mathplus$ waves in plants

F. Resentini
Primo
;
M. Grenzi
Secondo
;
L. Luoni;M.C. Bonza
Penultimo
Membro del Collaboration Group
;
A. Costa
Ultimo
Writing – Review & Editing
2021-01

Abstract

Calcium ions (Ca2+) play a key role in cell signaling across organisms. In plants, a plethora of environmental and developmental stimuli induce specific Ca2+ increases in the cytosol as well as in different cellular compartments including the endoplasmic reticulum (ER). The ER represents an intracellular Ca2+ store that actively accumulates Ca2+ taken up from the cytosol. By exploiting state-of-the-art genetically encoded Ca2+ indicators (GECIs), specifically the ER-GCaMP6-210 and R-GECO1, we report the generation and characterization of an Arabidopsis (Arabidopsis thaliana) line that allows for simultaneous imaging of Ca2+ dynamics in both the ER and cytosol at different spatial scales. By performing analyses in single cells, we precisely quantified i) the time required by the ER to import Ca2+ from the cytosol into the lumen; and ii) the time required to observe a cytosolic Ca2+ increase upon the pharmacological inhibition of the ER-localized type IIA Ca2+-ATPases. Furthermore, live imaging of mature, soil-grown plants revealed the existence of a wounding-induced, long-distance ER Ca2+ wave propagating in injured and systemic rosette leaves. This technology enhances high-resolution analyses of intracellular Ca2+ dynamics at the cellular level and in adult organisms and paves the way to develop new methodologies aimed at defining the contribution of subcellular compartments in Ca2+ homeostasis and signaling.
Settore BIO/04 - Fisiologia Vegetale
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/852604
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