Kornilova E S, Sorkin A D, Nikol'skiĭ N N
Tsitologiia. 1987 Aug;29(8):904-10.
Dynamics of compartmentalization of epidermal growth factor (EGF) in human carcinoma A431 cells during the first hour after initiation of endocytosis was examined by methods of the organelle fractionation on a 20% Percoll gradient and of the microfluorimetric visualization of endocytosis of rhodamine-labeled EGF (EGF-R). EGF was revealed in small vesicles localized in the peripheral region of cytoplasm in a few minutes after endocytosis initiation. During centrifugation in Percoll these vesicles (endosomes), with an average density of 1.038 g/ml, were seen co-sedimented with Golgi membranes. By one hour after initiation of endocytosis, EGF-R was accumulated in perinuclear zone, in a trans-Golgi region, as numerous big luminous centres that were apparently MB-endosomes and had the same density in Percoll as did small peripheral endosomes. Such centres appeared in several cells already within 5-10 minutes. In A431 cells EGF did not reach lysosomes within 60 minutes, because no accumulation of 125I-EGF was shown in lysosome corresponding regions of Percoll gradient (average density 1.070 g/ml).
采用在20% Percoll梯度上进行细胞器分级分离以及对罗丹明标记的表皮生长因子(EGF-R)内吞作用进行微荧光可视化的方法,研究了人癌A431细胞内吞作用开始后第一小时内表皮生长因子(EGF)的区室化动态变化。内吞作用开始几分钟后,在细胞质周边区域的小泡中发现了EGF。在Percoll中离心时,这些平均密度为1.038 g/ml的小泡(内体)与高尔基体膜一起沉降。内吞作用开始一小时后,EGF-R在核周区域、反式高尔基体区域积累,形成许多大的发光中心,这些中心显然是MB-内体,在Percoll中的密度与周边小内体相同。在5-10分钟内,几个细胞中就出现了这样的中心。在A431细胞中,EGF在60分钟内未到达溶酶体,因为在Percoll梯度的溶酶体相应区域(平均密度1.070 g/ml)未显示125I-EGF的积累。