Abstract
We expose methods for in vivo assessment of oxygen, nitric oxide (NO), and reactive oxygen species (ROS)/reactive nitrogen species (RNS), in the microcirculation during normoxia and hypoxia. We provide an example of the related mechanisms of ROS/RNS and oxygen level in the process of regulating capillary perfusion. Namely, we discuss the real time pO2 measurements in vivo in the microvessels and tissues of the hamster cheek pouch and window chamber preparations during normoxia and hypoxia, as well as the corresponding changes in ROS/RNS in systemic blood during normoxia and hypoxia under conditions where NO availability is maximally reduced.
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References
Haliwell B, Gutteridge JMC (1999) Free radical in Biology and Medicine. Oxford University Press, London
Dröge W (2001) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95
Halliwell B, Gutteridge JMC (1985) The chemistry of oxygen radicals and other oxygen-derived species. In: Free radicals in biology and medicine. Oxford University Press, New York, pp 20–64
Barreto JC, Smith GS, Stobel NHP, McQuillin PA, Miller TA (1995) Teraphtalic acid: a dosimeter for detection of hydroxyl radical in vitro. Life Sci 56:89–96
Parker L (ed) (1994) Oxygen radicals in biological systems. In: Methods in Enzymology, vol 234, part D. Academic, New York
Ignarro LJ (2002) Nitric oxide is an unique signaling molecule in the vascular system: a historical overview. J Physiol Pharmacol 53:503–514
Buerk DG (2007) Nitric oxide regulation of microvascular oxygen. Antioxid Redox Signal 9:829–843
Abu-Soud HM, Rousseau DL, Stuehr DJ (1966) Nitric oxide binding to the heme of neuronal nitric-oxide synthase links its activity to changes in oxygen tension. J Biol Chem 271:32515–32518
Bertuglia S, Giusti A (2005) The role of nitric oxide in capillary perfusion and oxygen delivery regulation during systemic hypoxia. Am J Physiol Heart Circ Physiol 288(2):H525–531
Shen W, Xu X, Ochoa M, Zhao G, Wolin MS, Hintze TH (1994) Role of nitric oxide in the regulation of oxygen consumption in conscious dogs. Circ Res 75:1086–1095
King CE, Melinyshyin MJ, Mewburn JD, Curtis SE, Winn MJ, Cain SM, Chapler CK (1994) Canine hindlimb flow and O2 uptake after inhibition of EDRF/NO synthesis. J Appl Physiol 76:1166–1171
Cabrales P, Tsai AG, Intaglietta M (2006) Nitric oxide regulation of microvascular oxygen exchange during hypoxia and hyperoxia. J Appl Physiol 100:1181–1187
Stamler JS (1995) S-nitrosothiols and the bioregulatory actions of nitrogen oxides through reactions with thiol groups. Curr Top Microbiol Immunol 196:19–36
Jia L, Bonaventura C, Bonaventura J, Stamler JS (1996) S-nitrosohaemoglobin: a dynamic activity of blood involved in vascular control. Nature 380:221–226
Intaglietta M, Johnson PC, Winslow RM (1996) Microvascular and tissue oxygen distribution. Cardiovasc Res 32:632–643
Hogan MC (1999) Phosphorescence quenching method for measurement of intracellular pO2 in isolated skeletal muscle fibers. J Appl Physiol 86:720–724
Kessler M, Harrison, DK, Hoper J (1986) Tissue oxygen measurement techniques. In: Baker CH, Nastuk WL, Orlando FL (eds) Microcirculatory technology. Academic, New York, pp. 391–425
Popel AS, Pittman RN, Ellsworth ML (1989) Rate of oxygen loss from arterioles is an order of magnitude higher than expected. Am J Physiol 256:H921–H924
Tsai AG, Johnson PC, Intaglietta M (2003) Oxygen gradients in the microcirculation. Physiol Rev 83:933–963
Kerger H, Saltzman DJ, Gonzales A, Tsai AG, van Ackern K, Winslow RM, Intaglietta M (1997) Microvascular oxygen delivery and interstitial oxygenation during sodium pentobarbital anesthesia. Anesthesiology 86:372–386
Pawlowski M, Wilson DF (1992) Monitoring of the oxygen pressure in the blood of live animals using the oxygen dependent quenching of phosphorescence. Adv Exp Med Biol 316:179–185
Sinaasappel M, van Iterson M, Ince C (1992) Microvascular oxygen pressure in the pig intestine during hemorrhagic shock and resuscitation. J Physiol (Lond) 514:245–253
Tsai AG, Friesenecker B, Mazzoni MC, Kerger H, Buerk DG, Johnson PC, Intaglietta M (1988) Microvascular and tissue oxygen gradients in the rat mesentery. Proc Natl Acad Sci USA 95:6590–6595
Wilson DF (1993) Measuring oxygen using oxygen dependent quenching of phosphorescence: a status report. Adv Exp Med Biol 333:225–232
Seylaz E, Pinard J (1977) Continuous intra-cerebral PO2 and PCO2 measurements by mass spectrometry: study of the influence of vasoactive drugs. Acta Neurol Scand 64:438–439
Grisham MB (1994) Oxidants and free radicals in inflammatory bowel disease. Lancet 344:859–861
Pryor WA, Stanley JP, Blair E (1976) Autoxidation of polyunsaturated fatty acids: II. A suggested mechanism for the formation of TBA-reactive materials from prostaglandin-like endoperoxides. Lipids 11:370–379
Bertuglia S, Reiter NJ (2007) Melatonin reduces ventricular arrhythmias and preserves capillary perfusion during ischemia-reperfusion events in cardiomyopathic hamsters. J Pineal Res 42(1):55–63
Salvemini D, Mazzon E, Dugo L, Serraino I, De Sarro A, Caputi AP, Cuzzocrea S (2001) Amelioration of joint disage in a rat model of collagen-induced arthritis in M40403, a superoxide dismutase mimetic. Arthritis Rheum 44:2909–2291
Szabo A, Hake P, Salzman AL, Szabo C (1999) Beneficial effects of mercaptothylguanidine, an inhibitor of the inducible isoform of NO synthase and a scavenger of peroxinitrite in a porcine model of delayed hemorrhagic shock. Crit Care Med 27:1343–1359
Halliwell B, Gutteridge JMC (2000) Detection of free radicals and other reactive species: trapping and finger printing. In: Halliwell B, Gutteridge JMC (eds) Free Radicals in Biology and Medicine. Oxford University Press, Oxford, pp 351–429
Morrow JD, Hill KE, Burk RF, Nammour TM, Badr KF, Roberts LJ (1990) A series of prostaglandin F2-like compounds are produced in vivo in humans by a non-cyclooxygenase, free radical-catalyzed mechanism. Proc Natl Acad Sci USA 87:9383–9387
Wink DA, Kim S, Coffin D, Cook JC, Vodovotz Y, Chistodoulou D, Jourd’heuil D, Grisham MB (1999) Detection of S-nitrosothiols by fluorometric and colorimetric methods. Methods Enzymol 301:201–211
Karlsson J (1997) Introduction to nutraology and radical formation. In: Antioxidants and exercise. Human Kinetics Press, Illinois, pp 1–143
Baker JE, Froncisz W, Joseph J, Kalyanaraman B (1997) Spin label oximetry to assess extracellular oxygen during myocardial ischemia. Free Radic Biol Med 22:109–115
Velan SS, Spencer RG, Zweier JL, Kuppusamy P (2000) Electron paramagnetic resonance oxygen mapping (EPROM): direct visualization of oxygen concentration in tissue. Magn Reson Med 43:804–809
Acworth IN, Bailey B (1997) Reactive oxygen species. In: The handbook of oxidative metabolism. ESA, Northampton, MA, pp 1–4.
Bertuglia S, Colantuoni A, Coppini G, Intaglietta M (1991) Hypoxia- or hyperoxia-induced changes in arteriolar vasomotion in skeletal muscle microcirculation. Am J Physiol 260:H362–H372
Bertuglia S, Giusti A, Del Soldato P (2004) Antioxidant activity of a nitro derivative of aspirin against ischemia reperfusion in hamster cheek pouch microcirculation. Am. J. Physiol. Gastrointestinal-Liver Physiol. 286(3):G437–G443
Bertuglia S, Giusti A (2003) Microvascular oxygenation, oxidative stress, nitric oxide suppression and superoxide dismutase during postischemic reperfusion. Am J Physiol 285:H1064–H1071
Cabrales P, Tsai AG, Intaglietta M (2004) Increased tissue PO2 and decreased O2 delivery and consumption after 80% exchange transfusionwith polymerized hemoglobin. Am J Physiol Heart Circ Physiol 287:H2825–H2833
Friesenecker B, Tsai AG, Dunser MW, Mayr AJ, Martini J, Knotzer H, Hasibeder W, Intaglietta M (2004) Oxygen distribution in the microcirculation following arginine vasopressin-induced arteriolar vasoconstriction. Am J Physiol Heart Circ Physiol 287:H1792–H1800
Torres Filho IP, Intaglietta M (1993) Microvessel PO2 measurement by phosphorence decay method. Am J Physiol Heart Circ Physiol 265:H1537–H1545
Kerger H, Groth G, Kalenka A, Vajkoczy P, Tsai AG, Intaglietta M (2003) PO2 measurements by phoroshorence quenching characteristics and applications of an automated system. Microvasc Res 15:93–101
Friedemann MN, Robinson SW, Gerhardt GA (1996) o-Phenylenediamine-modified carbon fiber electrodes for the detection of nitric oxide. Anal Chem 68:2621–2628
Cabrales P, Tsai AG, Intaglietta M (2004) Microvascular pressure and functional capillary density in extreme hemodilution with low and high plasma viscosity expanders. Am J Physiol Heart Circ Physiol 287:H363–H373
Tsai AG, Acero C, Nance PR, Frangos JA, Buerk DG, Intaglietta M (2005) Elevated plasma viscosity in extreme hemodilution increases perivascular nitric oxide concentration and microvascular perfusion. Am J Physiol Heart Circ Physiol 288:H1730–H1739
Sarelius IH (1968) Cell flow path influences transit time through striated muscle capillaries. Am J Physiol Heart Circ Physiol 250:H899–H907
Bohlen HG, Nase GP (2000) Dependence of intestinal arteriolar regulation on flow-mediated nitric oxide formation. Am J Physiol Heart Circ Physiol 279:H2249–H2258
Grishman MB, Jonshon GG, Lancaster JR (1966) Quantitation of nitrite and nitrate in extracellular fluids. Methods Enzymol 268:237–246
Golub AS, Barker MG, Pittman RN (2007) PO2 profiles near arterioles and tissue oxygen consumption in rat mesentery. Am J Physiol Heart Circ Physiol 293:H1097–H1106
Golub AS, Pittman RN (2008) PO2 measurements in the microcirculation using phosphorescence quenching microscopy at high magnification. Am J Physiol Heart Circ Physiol 294:H2095–H2916
Shibata M, Ichioka S, Ando J, Kamiya A (2001) Microvascular and interstitial PO2 measurements in rat skeletal muscle by phosphorence quenching. J Appl Physiol 91:321–327
Tarpey MM, Wink DA, Grishman MB (2008) Methods for detection of reactive metabolites of oxygen and nitrose: in vitro and in vivo considerations. Am J Physiol Regul Integr Comp Physiol 206:R431–R444
Halliwell B (1995) How to characterize an antioxidant: an update. Biochem Soc Symp 61:73–101
Rice-Evans CA, Diplock AT (1993) Current status of antioxidant therapy. Free Radic Biol Med 15:77–96
Skulachev VP (1997) Membrane-linked systems preventing superoxide formation. Biosci Rep. 17:347–366.
Prewitt RL, Johnson PC (1976) The effect of oxygen on arteriolar red cell velocity and capillary density in the rat cremaster muscle. Microvasc Res 12:59–70
Edmunds NJ, Marshall JM (2003) The roles of nitric oxide in dilating proximal and terminal arterioles of skeletal muscle during systemic hypoxia. J Vasc Res 40:68–76
Hangai-Hoger N, Tsai AG, Friesenecker B, Cabrales P, Intaglietta M (2005) Microvascular oxygen delivery and consumption following treatment with verapamil. Am J Physiol Heart Circ Physiol 288:H1515–H1520
Akaike T, Yoshida M, Miyamoto Y (1993) Antagonistic action of imidazolineoxyl N-oxides against endothelium-derived relaxing facto/NO through a radical reaction. Biochemistry 32:827–832
Christie MI, Griffith TM, Lewis MJ (1989) A comparison of basal and agonist-stimulated release of endothelium-derived relaxing factor from different arteries. Br J Pharmacol 98:397–406
Coste J, Vial JC, faury G, Deronzier A, Usson Y, Nicoud MR, Verdetti J (2002) NO synthesis, unlike respiration, influences intracellular oxygen tension. Biochem Biophys Res Comm 209:97–104
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Bertuglia, S., Intaglietta, M. (2010). pO2 and ROS/RNS Measurements in the Microcirculation in Hypoxia. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_2
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DOI: https://doi.org/10.1007/978-1-60761-411-1_2
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