Comparison of calcium, freeze-thaw, and Triton X-100 tegumental disruption/recovery techniques applied to Schistosoma mansoni.
Three techniques for the disruption/recovery of tegumental free-surface plasmalemma were compared by (i) morphological examination of carcasses and centrifugally-derived isolates, (ii) specific enrichment of bound surface tags (lectin) and of "marker" enzymes for membrane, and (iii) assessment of total protein and lectin recovered by each procedure. Procedures compared included the use of Triton X-100, freezing and thawing, and high ionic strength calcium. Triton X-100 consistently provided the greatest amounts of recovered surface membrane on a per worm basis, whereas calcium retained the highest amounts of alkaline p-nitrophenyl phosphatase, adenosine triphosphatase, and adenosine monophosphatase activity. Ultrastructural examination of membrane isolates and worm carcasses prepared by freezing and thawing indicated that significant amounts of parenchymal material contaminated the membrane fractions. Thus results based on the freeze-thaw technique can be difficult to interpret.
D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002122:Calcium Chloride; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D003208:Concanavalin A / Q000032:analysis; D005615:Freezing; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D009708:Nucleotidases / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols; D012550:Schistosoma mansoni / Q000032:analysis / Q000648:ultrastructure*
D008565:Membrane Proteins; D003208:Concanavalin A; D011092:Polyethylene Glycols; D017830:Octoxynol; D009708:Nucleotidases; D000469:Alkaline Phosphatase; D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases; D002122:Calcium Chloride