Red Blood Cell Homeostasis: Pharmacological Interventions to Explore Biochemical, Morphological and Mechanical Properties

Abstract

During their passage through the circulation, red blood cells (RBCs) encounter severe physiological conditions consisting of mechanical stress, oxidative damage and fast changes in ionic and osmotic conditions. In order to survive for 120 days, RBCs adapt to their surroundings by subtle regulation of membrane organization and metabolism. RBC homeostasis depends on interactions between the integral membrane protein band 3 with other membrane and cytoskeletal proteins, and with key enzymes of various metabolic pathways. These interactions are regulated by the binding of deoxyhemoglobin to band 3, and by a signaling network revolving around Lyn kinase and Src family kinase-mediated phosphorylation of band 3. Here we show that manipulation of the interaction between the lipid bilayer and the cytoskeleton, using various pharmacological agents that interfere with protein-protein interactions and membrane lipid organization, has various effects on: (1) morphology, as shown by high resolution microscopy and quantitative image analysis; (2) organization of membrane proteins, as indicated by immunofluorescence confocal microscopy and quantitative as well as qualitative analysis of vesicle generation; (3) membrane lipid organization, as indicated by flow cytometric analysis of phosphatidylserine exposure; (4) deformability, as assessed in capillary-mimicking circumstances using a microfluidics system; (5) deformability as determined using a spleen-mimicking device; (6) metabolic activity as indicated by metabolomics. Our data show that there is a complex relationship between red cell morphology, membrane organization and deformability. Also, our data show that red blood cells have a relatively high resistance to disturbance of membrane organization in vitro, which may reflect their capacity to withstand mechanical, oxidative and osmotic stress in vivo.

HbE/β-Thalassemia and Oxidative Stress: The Key to Pathophysiological Mechanisms and Novel Therapeutics

“…These changes in Band 3 then initiate alterations in the local membrane environment surrounding Band 3. The result of excessive phosphorylation of Band 3 protein is correlated with an enhanced membrane degradation generating the production of proinflammatory microparticles and increasing the rate of hemolysis (42,54,55,131).…”

“…It is when the b93 Cys are oxidized that Hb alterations occur, including Hb dimer/hemichrome formation, which can disrupt the functional Band 3 protein complex under conditions of oxidative stress (110). Hemolysis and RBC-derived microparticles in turn create and exacerbate vascular inflammation, including platelet activation and endothelial dysfunction (42). All these vascular consequences will decrease tissue perfusion and, under conditions where the inflammation is chronic, will likely lead to organ damage/ failure.…”

The Proteome of the Red Blood Cell: An Auspicious Source of New Insights into Membrane-Centered Regulation of Homeostasis

“…Data from labeling studies and pharmacological interventions in vitro suggest that a relationship between membrane organization and cytoplasmic protein association may not be restricted to band 3 and key glycolytic enzymes [15,16]. The signals that affect this association, their receptors, and their transduction pathways are largely unknown.…”

“…On the other hand, GSH and GSSG concentrations and acetyl-carnitine were found to be associated with the red blood cell aging process [17,18]. In both cases, alterations in membrane protein composition and organization are likely to be the primary cause of at least some of these changes [16,44]. …”

Реактивность Эритроцитов Крыс В Условиях Физической Нагрузки Разной Интенсивности

“…Особое значение в адаптивных реакциях организма к физическим нагрузкам принадлежит молодым эритроцитам, увеличение количества которых влияет на величину среднего диаметра эритроцитов и выполнение экстренного восполнения дефицита кислорода [1,2]. Исследование реакции красной крови на физическую нагрузку предполагает поэтапное включение различных механизмов, обеспечивающих кислородный запрос организма [3,4,5,6,7] и проявление способности противостоять механическому, окислительному и осмотическому стрессу in vivo [8]. Реакция организма к физическим нагрузкам сопровождается перераспределением резервов функциональных систем, реагирующих на экстремальную ситуацию [9,10].…”