A new tissue technique for evaluating effects of Bacillus thuringiensis toxins on insect midgut epithelium
A new tissue technique for evaluating effects of Bacillus thuringiensis toxins on insect midgut epithelium
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DOI：
10.1006/jipa.1996.4632
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发表时间：
1997-03-01
影响因子：
3.4
通讯作者：
Keddie, BA
中科院分区：
文献类型：
--
作者：
Braun, L;Keddie, BA
酶法去除粉纹夜蛾幼虫中肠的基膜和结缔组织后，得到上皮组织全菌群。将全菌群在人工血淋巴中营养，并使用活体染料try-pan蓝、吖啶橙子（AO）、碘化丙啶（PI）和4 '，6-二脒基-2-苯基吲哚（I）API）评估组织活力长达24小时。围食膜合成和苏云金芽孢杆菌Cry 1Ac原毒素对活性毒素的修饰证实了一些正常的上皮功能。使用AO和PI的组合进行活体染色，或DAPI揭示了用活化的Cry 1Ac毒素处理的组织的柱状上皮和再生细胞中的膜渗透性改变，而喂养和口服接种生物测定证实了Cry 1Ac毒性。选择DAPI以在快速和高度灵敏的测定中鉴定靶细胞。(C)北京：科学出版社.
Epithelial tissue wholemounts were produced after enzymatic removal of basal lamina and connective tissue from midguts of Trichoplusia ni larvae. Wholemounts were nourished in artificial hemolymph and tissue viability 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-phenylindole (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 AO and PI, or DAPI revealed altered membrane permeability in columnar epithelial and regenerative cells of tissues treated with activated Cry1Ac toxin while feeding and oral inoculation bioassays verified Cry1Ac toxicity. DAPI was selected to identify target cells in a rapid and highly sensitive assay. (C) 1997 Academic Press.