title: A Bright Fluorescent Probe for H2S Enables Analyte-Responsive, 3D Imaging in Live Zebrafish Using Light Sheet Fluorescence Microscopy.
abstract: Hydrogen sulfide (H2S) is a critical gaseous signaling molecule emerging at the center of a rich field of chemical and biological research. As our understanding of the complexity of physiological H2S in signaling pathways evolves, advanced chemical and technological investigative tools are required to make sense of this interconnectivity. Toward this goal, we have developed an azide-functionalized O-methylrhodol fluorophore, MeRho-Az, which exhibits a rapid >1000-fold fluorescence response when treated with H2S, is selective for H2S over other biological analytes, and has a detection limit of 86 nM. Additionally, the MeRho-Az scaffold is less susceptible to photoactivation than other commonly used azide-based systems, increasing its potential application in imaging experiments. To demonstrate the efficacy of this probe for H2S detection, we demonstrate the ability of MeRho-Az to detect differences in H2S levels in C6 cells and those treated with AOAA, a common inhibitor of enzymatic H2S synthesis. Expanding the use of MeRho-Az to complex and heterogeneous biological settings, we used MeRho-Az in combination with light sheet fluorescence microscopy (LSFM) to visualize H2S in the intestinal tract of live zebrafish. This application provides the first demonstration of analyte-responsive 3D imaging with LSFM, highlighting the utility of combining new probes and live imaging methods for investigating chemical signaling in complex multicellular systems.
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Hammers
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Taormina
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Cerda
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Leticia A
authors.initials: LA
authors.lastname: Montoya
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Seidenkranz
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Raghuveer
authors.initials: R
authors.lastname: Parthasarathy
authors.affiliation: †Department of Chemistry and Biochemistry, ‡Department of Physics, §Institute of Molecular Biology, ∥Materials Science Institute. University of Oregon, Eugene, Oregon 97403-1253, United States.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Pluth
mesh_terms: D000818:Animals; D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry*; D060326:Chemistry Techniques, Synthetic; D005452:Fluoresceins / Q000138:chemical synthesis / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006862:Hydrogen Sulfide / Q000032:analysis* / Q000737:chemistry*; D021621:Imaging, Three-Dimensional; D007814:Larva / Q000378:metabolism; D057230:Limit of Detection; D008856:Microscopy, Fluorescence / Q000379:methods*; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D001386:Azides; D005452:Fluoresceins; D005456:Fluorescent Dyes; C000622345:MeRho-Az compound; D006862:Hydrogen Sulfide