The ultrastructural localization of tri-n-butyltin in human erythrocyte membranes during shape transformation leading to hemolysis.
Scanning electron microscopy, transmission electron microscopy, freeze-fracture, and x-ray energy dispersive spectrometry were used to localize tri-n-butyltin (TBT) in human erythrocytes (RBC's). TBT induced a rapid shape transformation of the RBC discocyte to an echinocyte, which led to hemolysis at concentrations at or above 10 microM TBT. Electron dense spheres or ellipsoids were observed in association with blood cell membranes at or above 10 microM TBT. These structures were visualized initially in thin sections when postfixed with osmium tetroxide. Control cell preparations without TBT did not exhibit these structural densities when fixed with osmium. Freeze-fracture replicas confirmed the presence of TBT aggregates associated with cell membranes as intercalations in the lipid bilayer. In thin sections, these structures measured 71.5 +/- 18.2 nm in diameter. In freeze-fracture replicas of TBT-treated RBC's, particulate structures measuring 60 +/- 18.5 nm in diameter were present on membrane exoplasmic fracture faces and 59.6 +/- 10.8-nm depressions on membrane protoplasmic fracture faces. Qualitative x-ray energy dispersive spectrometry analysis of ultrathin sections of glutaraldehyde-carbohydrazide-embedded samples revealed that the membrane-associated aggregates contained tin. TBT-treated RBC's that were washed with normal saline resulted in a paucity of TBT aggregates associated with the membranes and a reduction in the RBC hemolysis rate. RBC shape transformation occurred at each concentration examined from 0.1 to 100 microM TBT, but was reversible below 1 microM TBT.
M
M
Porvaznik
B H
BH
Gray
D
D
Mattie
A G
AG
Jackson
R E
RE
Omlor
D004577:Electron Probe Microanalysis; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D004912:Erythrocytes / Q000166:cytology; D005614:Freeze Fracturing; D006461:Hemolysis / Q000187:drug effects*; D006651:Histocytochemistry; D006801:Humans; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D013194:Staining and Labeling; D014220:Trialkyltin Compounds / Q000097:blood* / Q000494:pharmacology
D014220:Trialkyltin Compounds; C011559:tributyltin