Abstract
Intravital microscopy is a powerful tool for evaluating vascular hyperpermeability in various vascular beds. Hemorrhagic shock after traumatic injury is known to induce microvascular hyperpermeability, life-threatening edema, and microcirculatory perfusion disturbances. Here we describe the microsurgical and imaging methods to study mesenteric vascular hyperpermeability using intravital microscopy, in a rat model of hemorrhagic shock. In this protocol, hemorrhagic shock is induced by controlled withdrawal of blood to reduce the mean arterial pressure (MAP) to 40 mmHg for 60 min, followed by resuscitation for 60 min. To study the changes in vascular permeability, the rats are given FITC-albumin, a fluorescent tracer, intravenously. The FITC-albumin flux across the vessel wall is measured in mesenteric postcapillary venules by determining intravascular and extravascular fluorescence intensity under intravital microscopy. Intravital microscopic evaluation of high molecular weight FITC-albumin permeability is a reliable indicator of microvascular hyperpermeability.
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References
Childs E et al (1999) Leukocyte adherence and sequestration following hemorrhagic shock and total ischemia in rats. Shock 11:248–252
Childs EW, Udobi KF, Wood JG, Hunter FA, Smalley DM, Cheung LY (2002) In vivo visualization of reactive oxidants and leukocyte-endothelial adherence following hemorrhagic shock. Shock 8:423–427
Tharakan B et al (2010) (-)-Deprenyl inhibits vascular hyperpermeability after hemorrhagic shock. Shock 33:56–63
Childs E et al (2007) Apoptotic signaling induces hyperpermeability following hemorrhagic shock. Am J Physiol Heart Circ Physiol 292:H3179–H3189
Childs E et al (2010) 17β-estradiol mediated protection against vascular leak after hemorrhagic shock: role of estrogen receptors and apoptotic signaling. Shock 34:229–235
Murao Y, Hata M, Ohnishi K, Okuchi K, Nakajima Y, Hiasa Y, Junger WG, Hoyt DB, Ohnishi T (2003) Hypertonic saline resuscitation reduces apoptosis and tissue damage of the small intestine in a mouse model of hemorrhagic shock. Shock 20:23–28
Childs EW, Udobi KF, Hunter FA, Dhevan V (2005) Evidence of transcellular albumin transport after hemorrhagic shock. Shock 23:565–570
Davidson MT, Deitch EA, Lu Q, Hasko G, Abungu B, Nemeth ZH, Zaets SB, Gaspers LD, Thomas AP, Xu DZ (2004) Trauma-hemorrhagic shock mesenteric lymph induces endothelial apoptosis that involves both caspase-dependent and caspase-independent mechanisms. Ann Surg 240:123–131
Therade-Matharan S, Laemmel E, Duranteau J, Vicaut E (2004) Reoxygenation after hypoxia and glucose depletion causes reactive oxygen species production by mitochondria in HUVEC. Am J Physiol Regul Integr Comp Physiol 287:R1037–R1043
Savoye G, Tamion F, Richard V, Varin R, Thuillez C (2005) Hemorrhagic shock resuscitation affects early and selective mesenteric artery endothelial function through a free radical-dependent mechanism. Shock 23:411–416
van Leeuwen ALI et al (2020) In vitro endothelial hyperpermeability occurs early following traumatic hemorrhagic shock. Clin Hemorheol Microcirc 75:121–133
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Williams, T.R., Childs, E.W. (2024). Evaluation of Mesenteric Microvascular Hyperpermeability Following Hemorrhagic Shock Using Intravital Microscopy. In: Tharakan, B. (eds) Vascular Hyperpermeability. Methods in Molecular Biology, vol 2711. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3429-5_4
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DOI: https://doi.org/10.1007/978-1-0716-3429-5_4
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-3429-5
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