Kisurina-Evgen'eva O P, Onishchenko G E
Department of Cytology and Histology, Biological Faculty, Moscow State University.
Tsitologiia. 2000;42(1):47-56.
In the present work the behavior of mitochondria and lysosomes during cell spreading has been investigated in normal conditions and under ATP-synthesis inhibitors: sodium aside and N,N-dicyclohexylcarbodiimide (DCCD). In the control culture, microtubules run along the stable edge and perpendicular to the leading edge in most of spreading cells. As a whole, microtubules form a dense network in these cells. However, the radial cells contain bundles of microtubules, radiating from the perinuclear area or form circular arrays around the nucleus. The microtubule network is more dense under inhibitory treatment, than in control conditions. In the control culture the spherical cells display numerous small mitochondria (staining with Rhodamine 123). In the process of cell spreading some elongated mitochondria appear, most of them being localized in the perinuclear area. The mitochondria of cells with radial microtubule organization are directed towards the cell periphery, while in cells with circular bundles of microtubules the mitochondria are localized chaotically. Under DCCD treatment the mitochondria retain the staining for 2-3 h. In the spreading cells, round mitochondria may be distributed all over the cytoplasm. In the presence of sodium aside the mitochondria are not stained. However, by means of phase contrast microscopy some disoriented thread-shaped structures are observed, obviously corresponding to mitochondria. In the control conditions, lysosomes (stained with Acridine orange) in spreading cells are dispersed chaotically, all over the cytoplasm, or are localized in the perinuclear area. In the presence of sodium aside lysosomes are observed only in the perinuclear area. Under DCCD treatment lysosomes do not accumulate the dye. Thus, the cytoskeleton modification and changes in the properties of membrane organelles, induced by ATP-synthesis inhibitors, do not prevent attachment, spreading or cell polarization.
在本研究中,我们研究了在正常条件下以及在ATP合成抑制剂(叠氮化钠和N,N-二环己基碳二亚胺(DCCD))作用下,细胞铺展过程中线粒体和溶酶体的行为。在对照培养物中,微管沿着大多数铺展细胞的稳定边缘并垂直于前沿排列。总体而言,微管在这些细胞中形成密集的网络。然而,放射状细胞含有从核周区域辐射出的微管束,或者在细胞核周围形成环状排列。与对照条件相比,在抑制处理下微管网络更密集。在对照培养物中,球形细胞显示出许多小线粒体(用罗丹明123染色)。在细胞铺展过程中,出现了一些细长的线粒体,其中大多数位于核周区域。具有放射状微管组织的细胞中的线粒体朝向细胞周边,而在具有环状微管束的细胞中,线粒体分布混乱。在DCCD处理下,线粒体保持染色2-3小时。在铺展细胞中,圆形线粒体可能分布在整个细胞质中。在存在叠氮化钠的情况下,线粒体不被染色。然而,通过相差显微镜观察到一些无定向的丝状结构,显然对应于线粒体。在对照条件下,铺展细胞中的溶酶体(用吖啶橙染色)在整个细胞质中混乱分散,或者位于核周区域。在存在叠氮化钠的情况下,仅在核周区域观察到溶酶体。在DCCD处理下,溶酶体不积累染料。因此,由ATP合成抑制剂诱导的细胞骨架修饰和膜细胞器性质的变化并不妨碍细胞附着、铺展或极化。