title: Cytoskeletal rearrangements in human red blood cells induced by snake venoms: light microscopy of shapes and NMR studies of membrane function.
abstract: RBCs (red blood cells) circulating through narrow blood capillaries withstand major deformation. The mechanical and chemical stresses commonly exerted on RBCs continue to attract interest for the study of membrane structure and function. Snake venoms are lethal biochemical 'cocktails' that often contain haemotoxins, metalloproteinases, myotoxins, neurotoxins, phosphodiesterases, phospholipases and proteases. We have monitored the effects of 4 snake venoms (Pseudechis guttatus, Oxyuranus scutellatus, Notechis scutatus and Naja kaouthia) on human RBCs using NMR spectroscopy, DIC (differential interference contrast) and confocal light microscopy. RBCs underwent reproducible stomatocytosis, with unusual geographical-like indentations, spherocytosis, followed by rapid lysis. Confocal micrographs using a fluorescent dye linked to phalloidin showed that the change in morphology was associated with the aggregation of actin in the cytoskeleton. (31)P NMR saturation transfer experiments recorded transport of the univalent anion HPA (hypophosphite) on a subsecond time scale, thereby reporting on the function of capnophorin or Band 3 linked to the cytoskeleton; anion-exchange activity was substantially reduced by venom treatment. We propose a molecular-cytological hypothesis for the shape and functional changes that is different from, or supplementary to, the more 'traditional' bilayer-couple hypothesis more often used to account for similar morphological changes invoked by other reagents.
authors.affiliation: School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia.
authors.forename: Tsz Wai
authors.initials: TW
authors.lastname: Yau
authors.forename: Rhiannon P
authors.initials: RP
authors.lastname: Kuchel
authors.forename: Jennifer M S
authors.initials: JM
authors.lastname: Koh
authors.forename: David
authors.initials: D
authors.lastname: Szekely
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Mirtschin
authors.forename: Philip W
authors.initials: PW
authors.lastname: Kuchel
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects; D000199:Actins / Q000378:metabolism; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D016824:Antigens, Human Platelet / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects* / Q000473:pathology; D048430:Cell Shape; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects* / Q000473:pathology; D005456:Fluorescent Dyes / Q000737:chemistry; D006461:Hemolysis; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D018613:Microscopy, Confocal; D010590:Phalloidine / Q000737:chemistry / Q000494:pharmacology; D012910:Snake Venoms / Q000494:pharmacology*
chemical_list: D000199:Actins; D001457:Anion Exchange Protein 1, Erythrocyte; D016824:Antigens, Human Platelet; D005456:Fluorescent Dyes; D012910:Snake Venoms; C083451:human platelet antigen 1b; D010590:Phalloidine