L. Braun et Ba. Keddie, A NEW TISSUE TECHNIQUE FOR EVALUATING EFFECTS OF BACILLUS-THURINGIENSIS TOXINS ON INSECT MIDGUT EPITHELIUM, Journal of invertebrate pathology, 69(2), 1997, pp. 92-104
Epithelial tissue wholemounts were produced after enzymatic removal of
basal lamina and connective tissue from midguts of Trichoplusia ni la
rvae. Wholemounts were nourished in artificial hemolymph and tissue vi
ability was assessed for up to 24 hr using the vital dyes try-pan blue
, acridine orange (AO), propidium iodide (PI), and 4',6-diamidino-2-ph
enylindole (I)API), Peritrophic membrane synthesis and modification of
Bacillus thuringiensis Cry1Ac protoxin to active toxin confirmed some
normal epithelial function, Vital staining using the combination of A
O and PI, or DAPI revealed altered membrane permeability in columnar e
pithelial and regenerative cells of tissues treated with activated Cry
1Ac toxin while feeding and oral inoculation bioassays verified Cry1Ac
toxicity. DAPI was selected to identify target cells in a rapid and h
ighly sensitive assay. (C) 1997 Academic Press.