Simultaneous intracellular chloride and pH measurements using a GFP-based sensor.
Chloride and protons perform important closely related roles in many cellular responses. Here we developed a ratiometric biosensor, ClopHensor, based on a highly chloride-sensitive Aequorea victoria GFP variant that is suited for the combined real-time optical detection of pH changes and chloride fluxes in live cells. We detected high chloride concentration in large dense-core exocytosis granules by targeting ClopHensor to these intracellular compartments.
Arosio
Daniele
D
National Enterprise for nanoScience and nanoTechnology, Istituto Nanoscienze, Consiglio Nazionale delle Ricerche and Scuola Normale Superiore, Pisa, Italy. daniele.arosio@cnr.it
Ricci
Fernanda
F
Marchetti
Laura
L
Gualdani
Roberta
R
Albertazzi
Lorenzo
L
Beltram
Fabio
F
D015374:Biosensing Techniques; D002460:Cell Line; D002712:Chlorides / Q000737:chemistry* / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000737:chemistry; D005089:Exocytosis; D014644:Genetic Variation; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence
D002712:Chlorides; D049452:Green Fluorescent Proteins