Londoño I, Bendayan M
Department of Anatomy, Faculty of Medicine, University of Montreal, Quebec, Canada.
J Histochem Cytochem. 1988 Aug;36(8):1005-14. doi: 10.1177/36.8.3392391.
We localized acidic glycoconjugates at the ultrastructural level by applying the enzyme-gold approach. Neuraminidase and hyaluronidase were adsorbed to colloidal gold particles and applied to tissue sections under optimal conditions for their enzymatic activity. Neuraminidase-gold labeling was distributed over the Golgi apparatus and associated secretory granules in exocrine pancreatic cells and duodenal goblet cells. Mitochondria were labeled over inner membranes. Labeling was also found over the dispersed chromatin in the nucleus. Plasma membranes, particularly the apical side, were labeled by gold particles. On the other hand, incubation of tissue sections with the hyaluronidase-gold complex resulted in intense labeling of the rER membranes, the plasma membrane, and the dense chromatin in the nucleus. Labeling was also found over the Golgi apparatus and associated secretory granules, but only in duodenal goblet cells. Specificity of the results was confirmed by various control experiments performed, indicating that the enzyme-gold technique is useful for detecting linked-sugar residues on tissue thin sections. Labelings found over intra- and extracellular compartments in the present work are discussed in light of previous biochemical indications as well as of other histochemical detections of these glycoconjugates.
我们通过应用酶-金法在超微结构水平上定位酸性糖缀合物。将神经氨酸酶和透明质酸酶吸附到胶体金颗粒上,并在其酶活性的最佳条件下应用于组织切片。神经氨酸酶-金标记分布在外分泌胰腺细胞和十二指肠杯状细胞的高尔基体及相关分泌颗粒上。线粒体内膜也有标记。在细胞核中分散的染色质上也发现了标记。质膜,特别是顶端一侧,有金颗粒标记。另一方面,用透明质酸酶-金复合物孵育组织切片导致粗面内质网(rER)膜、质膜和细胞核中致密染色质的强烈标记。在高尔基体及相关分泌颗粒上也发现了标记,但仅在十二指肠杯状细胞中。通过进行各种对照实验证实了结果的特异性,表明酶-金技术可用于检测组织薄片上的连接糖残基。根据先前的生化指标以及这些糖缀合物的其他组织化学检测结果,讨论了本研究中在细胞内和细胞外区室中发现的标记。