Disruption and removal of the tegument from Schistosoma mansoni with triton X-100.
Tegumental membranes of Schistosoma mansoni were disrupted by 0.2% Triton X-100 in Tris-maleate buffered/Kreb-Ringer's solution. Subsequent differential centrifugation of the disruption solution at 2,500 g and 30,000 g produced two pellets which contained membrane components. Examination of the carcass by scanning electron microscopy revealed that most of the exposed tegument of both male and female worms was removed, while surface membrane protected by close apposition of another surface (i.e., in the gynecophoral canal) remained intact. The parenchymal tissue (e.g., subtegumental muscle and tegumental perikarya), excretory and gut epithelia, and the tegument's basement membrane also remained intact. The selectivity of the disruption suggests that membrane in both pellets originated almost exclusively from the tegument. Although larger morphological features (i.e., surface crypts) present in the intact tegument did not maintain their form in the 2,500 g pellet, the high specific activity of 3H-concanavalin A retained by this fraction, and the presence of numerous spines and large pieces of membrane, suggest that the 2,500 g pellet contained most of the worm's disrupted surface membrane. Transmission electron microscopy demonstrated the presence of dense spinelike material and vesicles of various sizes and densities, as well as some mitochondria in the 30,000 g pellet. Low specific activity of 3H-concanavalin A in the post-30,000 g supernatant suggests that relatively few externally oriented, saccharide-containing molecules were solubilized from tegumental membranes by Triton X-100.
D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure; D002462:Cell Membrane / Q000648:ultrastructure; D002498:Centrifugation; D003208:Concanavalin A / Q000378:metabolism; D003593:Cytoplasm / Q000648:ultrastructure; D005260:Female; D008297:Male; D008566:Membranes / Q000648:ultrastructure; D008854:Microscopy, Electron; D017830:Octoxynol; D009940:Organoids / Q000648:ultrastructure; D011092:Polyethylene Glycols / Q000494:pharmacology*; D012550:Schistosoma mansoni / Q000648:ultrastructure*
D003208:Concanavalin A; D011092:Polyethylene Glycols; D017830:Octoxynol