Nist C, Von Der Mark K, Hay E D, Olsen B R, Bornstein P, Ross R, Dehm P
J Cell Biol. 1975 Apr;65(1):75-87. doi: 10.1083/jcb.65.1.75.
Three distinct antiprocollagen preparations were characterized and used in immunocytochemical staining of chick embryo corneal and tendon cells. The several ferritin-conjugated antibody preparations permitted similar location of procollagen in the cisternae of the rough endoplasmic reticulum and in Golgi elements in both cell types. The ability to demonstrate and interpret specific ferritin staining was dependent on the extent of membrane breakage in each of those organelles, coupled with adequate retention of cell morphology. Corneal fibroblasts appeared to suffer more extensive intracellular membrane damage under controlled conditions of homogenization than tendon fibroblasts, facilitating the identification of procollagen in Golgi vacuoles of these cells. None of the labeled material appeared to by cytoplasmic in origin since ferritin was observed in the cytoplasm only in the vicinity of Golgi elements that were extensively broken. This study extends previous immunological evidence for the presence of procollagen in the Golgi complex and calls attention to the problems to be encountered in locating the antigen in small Golgi vesicles and lamellae.
对三种不同的前胶原制剂进行了表征,并将其用于鸡胚角膜细胞和肌腱细胞的免疫细胞化学染色。几种铁蛋白偶联抗体制剂使两种细胞类型中前胶原在粗面内质网池和高尔基体中的定位相似。显示和解释特异性铁蛋白染色的能力取决于这些细胞器中每个细胞器的膜破裂程度,以及细胞形态的充分保留。在匀浆的受控条件下,角膜成纤维细胞似乎比肌腱成纤维细胞遭受更广泛的细胞内膜损伤,这有助于在这些细胞的高尔基体液泡中鉴定前胶原。由于仅在广泛破裂的高尔基体元件附近的细胞质中观察到铁蛋白,因此没有标记物质似乎起源于细胞质。本研究扩展了先前关于高尔基体复合物中存在前胶原的免疫学证据,并提请注意在小高尔基体囊泡和薄片中定位抗原时会遇到的问题。