Flickinger C J
J Cell Sci. 1978 Dec;34:53-63. doi: 10.1242/jcs.34.1.53.
The appearance of enzymic activity during the development of the Golgi apparatus was studied by cytochemical staining of renucleated amoebae. In cells enucleated for 4 days, there was a great decline in size and number of Golgi bodies, or dictyosomes. Subsequent renucleation by nuclear transplantation resulted in a regeneration of Golgi bodies. Samples of amoebae were fixed and incubated for cytochemical staining at intervals of 1, 6, or 24 h after renucleation. Enzymes selected for study were guanosine diphosphatase (GDPase), esterase, and thiamine pyrophosphatase (TPPase). All three were found in the Golgi apparatus of normal amoebae but they differed in their overall intracellular distribution. GDPase was normally present at the convex pole of the Golgi apparatus, in rough endoplasmic reticulum, and in the nuclear envelope. In amoebae renucleated for 1 h, light reaction product for GDPase was present throughout the small stacks of cisternae that represented the forming Golgi apparatus. By 6 h following the operation GDPase reaction product was concentrated at the convex pole of the Golgi apparatus. Esterase, which was distributed throughout the stacks of normal Golgi cisternae, displayed a similar distribution in the forming Golgi bodies as soon as they were visible. TPPase was normally present in the Golgi apparatus but was not found in the endoplasmic reticulum. In contrast to the other enzymes, TPPase reaction product was absent from the forming Golgi apparatus 1 and 6 h after renucleation, and did not appear in the Golgi apparatus until 24 h after operation. Thus, enzymes held in common between the rough endoplasmic reticulum and the Golgi apparatus were present in the forming Golgi apparatus as soon as it was detectable, but an enzyme cytochemically localized to the Golgi apparatus only appeared later in development of the organelle. It is suggested that Golgi membranes might be derived from the endoplasmic reticulum and thus immediately contain endoplasmic reticulum enzymes, while Golgi-specific enzymes are added later in development.
通过对去核变形虫进行细胞化学染色,研究了高尔基体发育过程中酶活性的出现情况。在去核4天的细胞中,高尔基体或高尔基体囊泡的大小和数量大幅下降。随后通过核移植重新植入细胞核导致高尔基体再生。在重新植入细胞核后1、6或24小时的间隔时间,对变形虫样本进行固定并孵育以进行细胞化学染色。选择用于研究的酶有鸟苷二磷酸酶(GDPase)、酯酶和硫胺素焦磷酸酶(TPPase)。这三种酶在正常变形虫的高尔基体中均有发现,但它们在细胞内的整体分布有所不同。GDPase通常存在于高尔基体的凸极、粗面内质网和核膜中。在重新植入细胞核1小时的变形虫中,代表正在形成的高尔基体的小囊泡堆叠中均存在GDPase的光反应产物。手术后6小时,GDPase反应产物集中在高尔基体的凸极。酯酶分布在正常高尔基体囊泡堆叠中,在可见的正在形成的高尔基体中也呈现类似分布。TPPase通常存在于高尔基体中,但在内质网中未发现。与其他酶不同,在重新植入细胞核后1小时和6小时,正在形成的高尔基体中没有TPPase反应产物,直到手术后24小时才出现在高尔基体中。因此,粗面内质网和高尔基体共有的酶在可检测到正在形成的高尔基体时就已存在,而细胞化学定位在高尔基体的酶则在该细胞器发育后期才出现。有人认为,高尔基体膜可能源自内质网,因此立即含有内质网酶,而高尔基体特异性酶则在发育后期添加。