Premium
Real‐time nitric oxide measurements in rat isolated hearts by 4,5‐diaminofluorescein fluorescence (698.11)
Author(s) -
Riess Matthias,
Bienengraeber Martin,
Cheng Qunli,
Keszler Agnes,
Weihrauch Dorothee
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.698.11
Subject(s) - nitric oxide , sodium nitroprusside , fluorescence , confocal microscopy , confocal , fluorescence microscope , veterans affairs , biophysics , chemistry , medicine , biology , microbiology and biotechnology , optics , physics
Real‐time measurement of nitric oxide (NO) before, during and after study interventions has numerous advantages over multiple endpoint measurements at various times. However, no reliable method has been described in Langendorff‐prepared hearts. Rat isolated hearts were loaded for 30 min with 20 µM of the fluorescent dye 4,5‐diaminofluorescein (DAF‐2) whose fluorescence (F) increases irreversibly in the presence of NO. Changes in NO were provoked by any combination of 30/120 min ischemia/reperfusion, the NO donor sodium nitroprusside and/or its scavenger cPTIO. Using a bifurcated fiberoptic probe left ventricular epicardial DAF‐2 F was measured online at 490/540 nm excitation/emission. Multiple regression analysis (alpha .05) was used to compare epicardial F after 185 min perfusion to confocal microscopy (488/550 nm) of transverse slices and to high‐performance liquid chromatography of tissue homogenate. Overall, all three measurements correlated significantly to each other and changed adequately with above treatments. Although online fluorescence lacks the spatial resolution of confocal microscopy it may provide a unique insight into real‐time effects of IR and potential cardioprotective treatments on the production of NO. This may serve as an important adjunct tool to investigate its role in studying cardioprotective strategies in isolated hearts. Supported by Department of Veterans Affairs (CARA‐026‐10F) and NIH. Grant Funding Source : Supported by Department of Veterans Affairs and NIH.