Khan K M, Hatfield J S, Drescher D G
Department of Otolaryngology, Wayne State University School of Medicine, Detroit, Michigan 48201.
Hear Res. 1991 Jun;53(2):223-9. doi: 10.1016/0378-5955(91)90056-f.
The luminal surface of the saccular macula in the rainbow trout is covered with a glycoconjugate-rich cell coat. The aim of this study was to identify specific carbohydrate moieties present in this coat, using biotinylated lectins as probes. Saccular tissues were fixed in Karnovsky's fixative for 2 h at 1-2 degrees C, followed by incubation with biotinylated lectins for 12-16 h at 25 degrees C. Lectin binding was visualized by performing avidin-biotin-peroxidase reactions. As controls, specimens were reacted with solutions of lectins preincubated with their specific inhibitory sugars. Staining was observed that was consistent with the presence of glucose, galactose, fucose, mannose, N-acetylglucosamine, N-acetylneuraminic acid, and N-acetylgalactosamine in the cell coat. The variability in the intensity of staining associated with the lectin-carbohydrate complexes suggests quantitative differences among the various carbohydrate moieties detected. The presence of these carbohydrates in the cell coat of the trout saccular macula also suggests biochemical similarities between cell coats in teleost and mammalian inner ear structures.
虹鳟鱼球囊斑的管腔表面覆盖着一层富含糖缀合物的细胞衣。本研究的目的是使用生物素化凝集素作为探针，鉴定该细胞衣中存在的特定碳水化合物部分。将球囊组织在卡诺夫斯基固定液中于1-2℃固定2小时，然后在25℃下与生物素化凝集素孵育12-16小时。通过进行抗生物素蛋白-生物素-过氧化物酶反应来观察凝集素结合情况。作为对照，将标本与预先用其特异性抑制糖孵育的凝集素溶液反应。观察到的染色结果与细胞衣中存在葡萄糖、半乳糖、岩藻糖、甘露糖、N-乙酰葡糖胺、N-乙酰神经氨酸和N-乙酰半乳糖胺一致。与凝集素-碳水化合物复合物相关的染色强度变化表明所检测的各种碳水化合物部分之间存在数量差异。这些碳水化合物在虹鳟鱼球囊斑细胞衣中的存在也表明硬骨鱼和哺乳动物内耳结构的细胞衣之间存在生化相似性。
