Lectin binding patterns to plasmalemmal glycoconjugates of goblet cells undergoing differentiation in vitro.
Lectin binding patterns to plasmalemmal glycoconjugates of goblet cells undergoing differentiation in vitro.
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凝集素与体外分化的杯状细胞质膜糖缀合物的结合模式。
DOI：
10.1002/jemt.1060160105
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Phillips,TE
中科院分区：
文献类型：
--
作者：
Frisch,EB;Phillips,TE
HT 29 - 18 N2（N2）细胞系的质膜糖缀合物在生长为（1）含葡萄糖培养基中未分化多层和（2）无葡萄糖培养基中获得杯状细胞表型的单层柱状细胞的细胞上进行表征。凝集素不能结合的薄片分离的N2细胞在没有固定。用醛固定后，可以看到凝集素结合位点的戏剧性暴露。当固定的单层细胞在包埋前染色时，通过抗生物素蛋白-生物素-复合过氧化物酶技术观察到的生物素化凝集素比胶体金偶联凝集素更有效。也可使用胶体金偶联凝集素对LR白色、Lowicryl K4 M和Lowicryl HM 20包埋的单层进行染色，检测凝集素结合位点。5种凝集素的结合（小麦胚、扁豆、花生、大豆和荆豆）的分化与分化阶段无关;“预分化”结果表明，在具有明显微绒毛、无粘液分泌颗粒或分泌颗粒很少的柱状细胞中，蓖麻凝集素的染色模式与具有大量分泌颗粒的分化良好的杯状细胞相同，蓖麻凝集素I是唯一一种凝集素的结合受细胞生长阶段影响的凝集素。差异化;刀豆（ConA）在包埋前对固定单层细胞的顶膜和基底膜进行了中度甚至均匀的标记，但包埋后的标记显示，所有柱状细胞的侧缘沿着有重度标记，顶膜和基底膜沿着有弱至中度的结合。
The plasmalemmal glycoconjugates of the HT29‐18N2 (N2) cell line were characterized on cells grown as (1) undifferentiated multilayers in glucose‐containing culture media and (2) monolayers of columnar cells acquiring the goblet cell phenotype in glucose‐free media. Lectins were unable to bind sheets of detached N2 cells in the absence of fixation. Following fixation with aldehydes, a dramatic unmasking of lectin binding sites was seen. When fixed monolayers were stained prior to embedding, biotinylated lectins, visualized by the avidin‐biotin‐complexed peroxidase technique, were more efficient than collodial gold‐coupled lectins. Lectin binding sites could also be detected by using collodial gold‐coupled lectins to stain monolayers embedded in LR White, Lowicryl K4M, and Lowicryl HM20. The binding of 5 lectins (wheat germ,Dolichos bifluros, peanut, soybean, andUlex europeus) was found to be independent of the stage of differentiation; “pre‐differentiated” columnar cells which had prominent microvilli and no or few mucous secretory granules had identical staining patterns as well‐differentiated goblet cells with large numbers of secretory granules.Ricinus communis Iwas the only lectin whose binding was influenced by the stage of differentiation; it intensely labeled undifferentiated multilayers of N2 cells but only weakly labeled basolateral membranes of differentiated monolayers.Canavalia ensiformas(ConA) caused a moderate and even labeling of both apical and basolateral membranes of fixed monolayers stained prior to embedding, but post‐embedding labeling revealed heavy labeling along the lateral margins of all columnar cells and weak to moderate binding along the apical and basal cell surface.
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