Leakage responses to L - NAME differ with the fluorescent dye used to label albumin ROLANDO

Rumbaut, Rolando E., Norman R. Harris, Arshad J. Sial, Virginia H. Huxley, and D. Neil Granger. Leakage responses to L-NAME differ with the fluorescent dye used to label albumin. Am. J. Physiol. 276 (Heart Circ. Physiol. 45): H333–H339, 1999.—Nitric oxide synthase (NOS) inhibitors have been reported to increase as well as to decrease microvascular transport of macromolecules in a variety of models. This study was performed to determine whether the influence of NOS inhibition on albumin leakage was dependent on the fluorescent dyes used to label albumin. Albumin leakage was assessed in rat mesenteric venules during control conditions and after exposure to the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME). Albumin was labeled with any one of four dyes: FITC, sulforhodamine 101 [Texas Red (TR)], dichlorotriazinyl aminofluorescein (DTAF), or Oregon Green 514 (OG). Superfusion with L-NAME (1024 M) was accompanied by an increase in leakage of FITC-labeled albumin (n 5 12) but not of albumin labeled with DTAF (n 5 10), TR (n 5 10), or OG (n 5 4). In vessels perfused with both FITCand TR-labeled albumin (n 5 12), superfusion with L-NAME increased leakage of FITCbut not TR-labeled albumin. In conclusion, albumin leakage responses to L-NAME differ among various fluorescent dyes. Therefore, caution is advised in comparison of albumin leakage results that utilize different fluorescent dyes.