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四膜虫细胞中的凝集素结合位点。II. 主要针对非分泌成分的标记分析。

Lectin binding sites in Paramecium tetraurelia cells. II. Labeling analysis predominantly of non-secretory components.

作者信息

Lüthe N, Plattner H

出版信息

Histochemistry. 1986;85(5):377-88. doi: 10.1007/BF00982667.

Abstract

All the lectin-FITC conjugates tested (ConA, RCA II, WGA) bind to the surface of Paramecium cells. Yet only WGA yields a distinct fluorescent pattern; it contours the basis of cilia and in some cells it brilliantly stains a few neighbouring rows of the regular surface fields in the anterioventral region (a region known to contain extensive fields of linear aggregates of freeze-fracture particles and to be engaged in conjugation). Incubation in vivo with WGA-FITC resulted in the selective labeling of the cytopharyngeal region as well as of the cytoproct. On Lowicryl K4M sections, WGA-gold probes concomitantly labeled disk-shaped vesicles that are assumed in the literature to serve as shuttle vesicles between these two cell regions and, thus, to connect forming and defecating digesting vacuoles (stages DV I and DV IV). On K4M sections WGA-Au stains also most other components of the lysosomal system. Also on K4M sections RCA II-Au labeled the walls of bacteria contained in DV I and II type digesting vacuoles (but not lysosomes identified bona fide by their size and shape and by their frequent vicinity to or continuity with digesting vacuoles). The WGA data largely support previous conclusions on the possible functional connection of all these elements (DV I-IV, smaller lysosomes, disk-shaped vesicles etc.) of the lysosomal system in Paramecium, as proposed by Allen and his group on the basis of other lines of evidence. As shown in the accompanying paper, ConA-FITC stained ghosts (formed after massive trichocyst exocytosis) also abut into DV-like structures. The different results obtained with the three lectins tested reflect the complex sorting machinery contained in the elaborate lysosomal system of a Paramecium cell. In the cytosol, finally, there occurs a particularly intense staining with ConA-gold, applied to Lowicryl sections, that probably represents glycogen-like particles. The same procedure reveals some weak staining of secretory contents and of nuclear structures.

摘要

所有测试的凝集素 - FITC 缀合物(刀豆球蛋白 A、蓖麻凝集素 II、小麦胚凝集素)都能与草履虫细胞表面结合。然而,只有小麦胚凝集素产生明显的荧光图案;它勾勒出纤毛基部的轮廓,在一些细胞中,它还能强烈地染色前腹区域几排相邻的规则表面区域(该区域已知含有大量冷冻断裂颗粒的线性聚集体,并且参与接合过程)。用 WGA - FITC 进行体内孵育导致胞咽区域以及胞肛的选择性标记。在 Lowicryl K4M 切片上,WGA - 金探针同时标记了圆盘状小泡,文献中认为这些小泡在这两个细胞区域之间充当穿梭小泡,从而连接形成和排泄消化液泡(DV I 和 DV IV 阶段)。在 K4M 切片上,WGA - Au 也能染色溶酶体系统的大多数其他成分。同样在 K4M 切片上,RCA II - Au 标记了 DV I 和 II 型消化液泡(但不是通过其大小、形状以及与消化液泡的频繁相邻或连续性而确定的真正溶酶体)中所含细菌的壁。WGA 的数据在很大程度上支持了 Allen 及其团队基于其他证据所提出的关于草履虫溶酶体系统中所有这些元素(DV I - IV、较小的溶酶体、圆盘状小泡等)可能的功能联系的先前结论。如随附论文所示,ConA - FITC 染色的幽灵结构(在大量刺丝泡胞吐作用后形成)也与 DV 样结构相邻。用三种测试凝集素获得的不同结果反映了草履虫细胞复杂的溶酶体系统中包含的复杂分选机制。最后,在应用于 Lowicryl 切片的 ConA - 金染色中,胞质溶胶中出现特别强烈的染色,这可能代表类糖原颗粒。相同的程序还显示出分泌内容物和核结构的一些弱染色。

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