INTRACELLULAR LOCALIZATION AND QUANTIFICATION OF HYDROGEN SULFIDE AND NITROGEN DIOXIDE

Abstract

DETECTION OF SMALL GASEOUS MOLECULES USING FLUORESCENT PROBES ALLOWS TO COUPLE QUANTITATIVE INFORMATION OBTAINED USING FLUORESCENT PROBES WITH HIGH QUALITY CONFOCAL IMAGING TO UNRAVEL THE INTRACELLULAR LOCALIZATION OF THE GASEOUS MOLECULES. THIS STUDY WAS BASED ON TWO FLUORESCENT PROBES WHICH WERE HYDROPHOBIC IN NATURE, HENCE A NOVEL CATIONIC LIPOSOME MEDIATED DELIVERY METHOD WAS UTILISED FOR SUBCELLULAR DETECTION. THE NO2 SPECIFIC PROBE WAS INVESTIGATED IN CELLULAR ENVIRONMENT IN THE PRESENCE OF DEANO TO DETERMINE THE FORMATION OF NO2 ENDOGENOUSLY PRODUCED NO WAS STIMULATED BY LIPOPOLYSACCHARIDES (LPS) AND INCREASE IN FLUORESCENT INTENSITY WAS OBSERVED. THE H2S SPECIFIC PROBE WAS ANALYSED FOR DOSE DEPENDENT INCREASE IN FLUORESCENT INTENSITY IN RAW 264.7 CELLS USING EXTERNAL DONORS SUCH AS NA2S AND NAPS. THE ENDOGENOUS H2S WAS MONITORED IN HEK 293 CELLS USING GENE KNOCKDOWN AND OVEREXPRESSION OF INHERENT CYSTATHIONINE GAMMA-LYASE GENE. FINALLY, A COMBINATION OF NO2 SENSITIV