title: Imaging of calcium dynamics in pollen tube cytoplasm.
abstract: Cytoplasmic calcium [(Ca(2+))cyt] is a central component of cellular signal transduction pathways. In plants, many external and internal stimuli transiently elevate (Ca(2+))cyt, initiating downstream responses that control different features of plant development. In pollen tubes the establishment of an oscillatory gradient of calcium at the tip is essential for polarized growth. Disruption of the cytosolic Ca(2+) gradient by chelators or channel blockers inhibits pollen tube growth. To quantify the physiological role of (Ca(2+))cyt in cellular systems, genetically encoded Ca(2+) indicators such as Yellow Cameleons (YCs) have been developed. The Cameleons are based on a fluorescence resonance energy transfer (FRET) process. Here, we describe a method for imaging cytoplasmic Ca(2+) dynamics in growing pollen tubes that express the fluorescent calcium indicator Yellow Cameleon 3.6 (YC 3.6), using laser-scanning confocal microscopy.
authors.affiliation: Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Dr. Héctor Torres (INGEBI-CONICET), Vuelta de Obligado 2490, Buenos Aires, C1428ADN, Argentina.
authors.forename: María Laura
authors.initials: ML
authors.lastname: Barberini
authors.forename: Jorge
authors.initials: J
authors.lastname: Muschietti
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D002118:Calcium / Q000502:physiology*; D002147:Calmodulin / Q000378:metabolism; D016764:Cell Polarity; D003470:Culture Media; D003600:Cytosol / Q000378:metabolism / Q000648:ultrastructure; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D018525:Germination; D008164:Luminescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods; D053205:Pollen Tube / Q000378:metabolism / Q000648:ultrastructure*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D002147:Calmodulin; D003470:Culture Media; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C579281:YCaM3.6 protein; D002118:Calcium