|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Cytoskeletal rearrangements in human red blood cells induced by snake venoms: light microscopy of shapes and NMR studies of membrane function
Tsz Wai Yau*†, Rhiannon P. Kuchel‡, Jennifer M. S. Koh*, David Szekely§, Peter J. Mirtschin¶ and Philip W. Kuchel1*‖
*School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia, †Australian Nuclear Science and Technology Organisation, Lucas Heights, Sydney, NSW 2232, Australia, ‡Department of Biological Sciences, Macquarie University, Sydney, NSW 2109, Australia, §Mark Cowley Lidwill Program in Cardiac Electrophysiology, The Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia, ¶School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA 5001, Australia, and ‖Singapore Bioimaging Consortium, BioMedical Sciences Institutes, Biopolis, Singapore 138667, Singapore
1To whom correspondence should be addressed (email email@example.com).
Movie 1: Microscopy recorded time course of RBC morphology evolution after P. guttatus venom (100 µg/ml) treatment at 25°C
The corresponding time is indicated on the top left-hand corner of the video (in the format of h:m:s). The video showed that the venom induced a systematic RBC shape evolution before haemolysis. Initially RBCs were observed as discocytes, which then rapidly evolved into stomatocytes following the addition of snake venom. Subsequently, the RBCs developed into stomatocytes with unusual geographical like indentations, then spherocytes, and ultimately lysis. As observed from the video, cells that did not undergo spherocytosis reverted back into stomatocytes, and some further reverted back to discocyte like shapes. Cells that had completed spherocytosis were seen to lyse rapidly after.
Movie 2: Microscopy recorded time course of RBC morphology evolution after PLA2 (5 U/ml, from honey bee venom) treatment at 37°C
The corresponding time is indicated on the top left-hand corner of the video (in the format of h:m:s). Upon PLA2 treatment, the cell morphology of the RBCs changed from discocytes to various stages of echinocytes, followed by spherocytes and ultimately haemolysis. The following are examples of RBCs which underwent PLA2-induced haemolysis: (1) RBCs that evolved into stomatocytes at 3 h 17 min, echinocytes at 3 h 56 min then rapidly changed into spherocytes, and ultimately haemolysis occurred at 5 h 34 min; (2) RBCs that evolved into echinocytes of various stages (Type 1, at 1 h; Type 2, at 1 h 30 min; Type 3, at 1 h 47 min; and Types 4 and 5, at 4 h 30 min) and then spherocytes at 5 h 18 min and ultimately haemolysis at 7 h 15 min.
Movie 3: Time course of light micrographs of the evolution of RBC morphology after P. guttatus venom (100 µg/ml) treatment in the presence of Complete™ protease inhibitor (7×) at 25°C
The corresponding time is indicated on the top left-hand corner of the video panel (in the format of h:m:s). The video shows that the RBCs were not affected by the venom; the RBCs remained as discocytes, suggesting that the Complete™ protease inhibitor inhibited the haemolyic activity of the venom.
Received 8 January 2011/7 July 2011; accepted 21 September 2011
Published as Cell Biology International Immediate Publication 21 September 2011, doi:10.1042/CBI20110012
© The Author(s) Journal compilation © 2012 Portland Press Limited