Lectin binding sites in Paramecium tetraurelia cells. II. Labeling analysis predominantly of non-secretory components.
Lectin binding sites in Paramecium tetraurelia cells. II. Labeling analysis predominantly of non-secretory components.
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草履虫细胞中的凝集素结合位点。
DOI：
10.1007/bf00982667
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
H. Plattner
中科院分区：
文献类型：
--
作者：
N. Lüthe;H. Plattner
所有测试的凝集素-FITC缀合物（ConA、RCA II、WGA）结合草履虫细胞的表面。然而，只有WGA产生一个独特的荧光图案;它的轮廓纤毛的基础，并在某些细胞中，它明亮地染色的几个相邻行的规则的表面领域的前腹区域（一个区域已知包含广泛的领域的线性聚集体的冷冻断裂颗粒和从事共轭）。温育在体内与WGA-FITC导致的选择性标记的cytopharyngeal区域以及cytoproct。在Lowicryl K4M切片上，WGA-金探针同时标记了文献中假定用作这两个细胞区域之间穿梭囊泡的盘状囊泡，因此连接了形成和排便消化空泡（DV I和DV IV期）。在K4M切片上，WGA-Au也染色溶酶体系统的大多数其他组分。此外，在K4M切片上，RCA II-Au标记了DV I型和II型消化空泡中所含细菌的壁（但不是通过其大小和形状以及其与消化空泡的频繁邻近或连续性而真正鉴定的溶酶体）。WGA数据在很大程度上支持了先前关于所有这些元件（DV I-IV，较小的溶酶体，盘状囊泡等）可能的功能连接的结论。艾伦和他的小组根据其他证据提出的草履虫中溶酶体系统。如随附的论文所示，ConA-FITC染色的血影（在巨大的包囊胞吐后形成）也邻接DV样结构。用三种凝集素测试得到的不同结果反映了草履虫细胞中复杂的溶酶体系统所包含的复杂分选机制。最后，在细胞质中，应用于Lowicryl切片的ConA-金染色特别强烈，可能代表糖原样颗粒。同样的程序显示了一些弱染色的分泌内容物和核结构。
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.