title: Hemolysis is a primary ATP-release mechanism in human erythrocytes.
abstract: The hypothesis that regulated ATP release from red blood cells (RBCs) contributes to nitric oxide-dependent control of local blood flow has sparked much interest in underlying release mechanisms. Several stimuli, including shear stress and hypoxia, have been found to induce significant RBC ATP release attributed to activation of ATP-conducting channels. In the present study, we first evaluated different experimental approaches investigating stimulated RBC ATP release and quantifying hemolysis. We then measured ATP and free hemoglobin in each and every RBC supernatant sample to directly assess the contribution of hemolysis to ATP release. Hypotonic shock, shear stress, and hypoxia, but not cyclic adenosine monophosphate agonists, significantly enhanced ATP release. It tightly correlated, however, with free hemoglobin in RBC supernatants, indicating that lysis was responsible for most, if not all, ATP release. Luminescence ATP imaging combined with simultaneous infrared cell imaging showed that ATP was released exclusively from lysing cells with no contribution from intact cells. In summary, with all stimuli tested, we found no evidence of regulated ATP release from intact RBCs other than by cell lysis. Such a release mechanism might be physiologically relevant in vivo, eg, during exercise and hypoxia where intravascular hemolysis, predominantly of senescent cells, is augmented.
authors.affiliation: Centre de Recherche, Centre Hospitalier de l'Université de Montréal, Montreal, QC, Canada;
authors.forename: Jacek
authors.initials: J
authors.lastname: Sikora
authors.affiliation: Centre de Recherche, Centre Hospitalier de l'Université de Montréal, Montreal, QC, Canada; Department of Medicine, Université de Montréal, Montreal, QC, Canada; and.
authors.forename: Sergei N
authors.initials: SN
authors.lastname: Orlov
authors.affiliation: Department of Physiology, Graduate School of Medicine, Nagoya University, Nagoya, Japan.
authors.forename: Kishio
authors.initials: K
authors.lastname: Furuya
authors.affiliation: Centre de Recherche, Centre Hospitalier de l'Université de Montréal, Montreal, QC, Canada; Department of Medicine, Université de Montréal, Montreal, QC, Canada; and.
authors.forename: Ryszard
authors.identifier: http://orcid.org/0000-0002-9798-7651
authors.initials: R
authors.lastname: Grygorczyk
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D015687:Cell Hypoxia; D000242:Cyclic AMP / Q000819:agonists; D004912:Erythrocytes / Q000378:metabolism* / Q000473:pathology; D006461:Hemolysis; D006801:Humans; D009997:Osmotic Pressure; D033081:Shear Strength; D013312:Stress, Physiological / Q000187:drug effects
chemical_list: D000255:Adenosine Triphosphate; D000242:Cyclic AMP
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