title: Measuring spatial and temporal Ca2+ signals in Arabidopsis plants.
abstract: Developmental and environmental cues induce Ca(2+) fluctuations in plant cells. Stimulus-specific spatial-temporal Ca(2+) patterns are sensed by cellular Ca(2+) binding proteins that initiate Ca(2+) signaling cascades. However, we still know little about how stimulus specific Ca(2+) signals are generated. The specificity of a Ca(2+) signal may be attributed to the sophisticated regulation of the activities of Ca(2+) channels and/or transporters in response to a given stimulus. To identify these cellular components and understand their functions, it is crucial to use systems that allow a sensitive and robust recording of Ca(2+) signals at both the tissue and cellular levels. Genetically encoded Ca(2+) indicators that are targeted to different cellular compartments have provided a platform for live cell confocal imaging of cellular Ca(2+) signals. Here we describe instructions for the use of two Ca(2+) detection systems: aequorin based FAS (film adhesive seedlings) luminescence Ca(2+) imaging and case12 based live cell confocal fluorescence Ca(2+) imaging. Luminescence imaging using the FAS system provides a simple, robust and sensitive detection of spatial and temporal Ca(2+) signals at the tissue level, while live cell confocal imaging using Case12 provides simultaneous detection of cytosolic and nuclear Ca(2+) signals at a high resolution.
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University; zhu148@purdue.edu.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Bindley Bioscience Center, Purdue University.
authors.forename: Aaron
authors.initials: A
authors.lastname: Taylor
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University.
authors.forename: Shenyu
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University; Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences.
authors.forename: Dayong
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University; College of Environmental & Resource Science, Zhejiang University.
authors.forename: Ying
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University; Dryland Agriculture Research Centre, Shanxi Academy of Agricultural Sciences.
authors.forename: Gaimei
authors.initials: G
authors.lastname: Liang
authors.affiliation: Department of Horticulture and Landscape Architecture, Purdue University; Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences.
authors.forename: Jian-Kang
authors.initials: JK
authors.lastname: Zhu
mesh_terms: D000331:Aequorin / Q000737:chemistry; D017360:Arabidopsis / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D020013:Calcium Signaling; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D030821:Plants, Genetically Modified; D036226:Seedlings / Q000737:chemistry / Q000166:cytology / Q000378:metabolism
chemical_list: D000331:Aequorin; D002118:Calcium