A novel method of measuring nitric-oxide-dependent fluorescence using 4,5-diaminofluorescein (DAF-2) in the isolated Langendorff-perfused rabbit heart

Pflügers Archiv : European journal of physiology
Vanlata H PatelG Andre Ng

Abstract

4,5-Diaminofluorescein (DAF-2) has been used to measure nitric oxide (NO) activity from a variety of preparations. The aim of this study was to develop a method to assess changes in NO fluorescence using DAF-2 in isolated rabbit hearts (2.0-2.5 kg, n = 8). Hearts were perfused in constant flow Langendorff mode and instrumented to record aortic perfusion pressure, left ventricular pressure and left ventricular epicardial fluorescence using a bifurcated light guide at excitation wavelengths of 470 +/- 10, 480 +/- 10, 490 +/- 10 and 500 +/- 10 nm collected at 535 nm. DAF-2 DA was loaded using a single bolus 150-microl (1 micromol) injection. Changes in NO-dependent fluorescence were determined using the NO donor sodium nitroprusside (SNP: 100 microM), NO-dependent vasodilator bradykinin (BK: 100 microM) and non-specific NO synthase inhibitor NG-nitro-L-arginine (LNA: 200 microM) before and after loading hearts with DAF-2 DA. Before loading, these agents did not alter epicardial fluorescence. After loading, SNP, BK and LNA produced a consistent change in each excitation wavelength. Together, this suggests that change in fluorescence represents changes in the level of NO. SNP and BK increased whilst LNA significantly decreased left ...Continue Reading

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Citations

Jul 20, 2010·Annual Review of Analytical Chemistry·Evan M Hetrick, Mark H Schoenfisch
May 29, 2012·American Journal of Physiology. Heart and Circulatory Physiology·Ronglih LiaoChee Chew Lim
Oct 31, 2012·Vascular Pharmacology·Tsutomu YamashitaJunichiro Yamamoto
Jul 18, 2016·Autonomic Neuroscience : Basic & Clinical·G André Ng
Dec 24, 2017·Journal of Neuroinflammation·Yoshinori HaraguchiAkira Monji
Jul 24, 2013·The Journal of Physiology·J H Coote
Sep 10, 2013·Experimental Physiology·G André Ng
Apr 4, 2015·Experimental Physiology·Kieran E Brack
Mar 19, 2021·Scientific Reports·Toru Murakawa-HirachiAkira Monji
Mar 2, 2018·Environmental Science & Technology·Rikke PoulsenMartin Hansen

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