REGULATION OF CYTOSKELETAL PROTEINS INVOLVED IN CELL CONTACT FORMATION DURING DIFFERENTIATION OF GRANULOSA-CELLS ON EXTRACELLULAR-MATRIX
REGULATION OF CYTOSKELETAL PROTEINS INVOLVED IN CELL CONTACT FORMATION DURING DIFFERENTIATION OF GRANULOSA-CELLS ON EXTRACELLULAR-MATRIX
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DOI：
10.1073/pnas.83.9.2894
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发表时间：
1986-05-01
影响因子：
11.1
通讯作者：
AMSTERDAM, A
中科院分区：
文献类型：
--
作者：
BENZEEV, A;AMSTERDAM, A
在颗粒细胞分化过程中，在细胞外基质（ECM）包被的培养板上分析了参与确定细胞接触和形状的细胞骨架蛋白的组织和表达。大鼠排卵前卵泡颗粒细胞在ECM上呈上皮样，相邻细胞间形成大量缝隙连接。这些细胞有很少的肌动蛋白电缆，往往只有一个弥漫性的肌动蛋白和少量的黏着斑在非常薄的粘着斑网站。与此相反，塑料上生长的细胞形成了单层的扁平细胞，间隙连接的数量减少，但有许多应力纤维和丰富的大的含纽蛋白的焦点接触。在ECM上，细胞被刺激产生高水平的孕酮，而在塑料培养皿上的细胞中只有微量的类固醇积累。[35 S]甲硫氨酸标记的细胞的二维凝胶电泳分析显示黏着斑蛋白α-与在塑料上生长的细胞相比，在ECM上生长的细胞中的肌动蛋白和肌动蛋白合成，而微管蛋白和中间丝蛋白波形蛋白的合成保持不变。RNA印迹分析显示，ECM板细胞中肌动蛋白mRNA水平显著降低，而微管蛋白mRNA水平基本保持不变。它的结论是，在体外ECM颗粒细胞的分化与细胞形状和细胞接触的变化，并参与确定这些细胞结构的细胞骨架蛋白的组织和表达的这种变化。
The organization and the expression of cytoskeletal proteins involved in determining cell contact and shape were analyzed in granulosa cells during their differentiation on extracellular matrix (ECM)-coated culture plates. Rat granulosa cells from preovulatory follicles displayed an epithelial shape on ECM and formed multilayered aggregates with numerous gap junctions between neighboring cells. These cells had few actin cables and often only a diffuse pattern of actin and a low amount of vinculin in very thin focal adhesion sites. In contrast, cells grown on plastic formed a monolayer of flat cells with a reduced number of gap junctions but with numerous stress fibers and abundant large vinculin-containing focal contacts. On ECM, the cells were stimulated to produce high levels of progesterone, while only trace amounts of the steroid accumulated in cells on plastic dishes. Two-dimensional gel electrophoretic analysis of [35S]methionine-labeled cells revealed a dramatic decrease in vinculin, .alpha.-actinin, and actin synthesis in cells grown on ECM, as compared to cells grown on plastic, while the synthesis of the tubulins and of the intermediate filament protein vimentin remained unchanged. RNA blot analysis showed a marked decrease in actin mRNA levels in cells from ECM plates, while the level of tubulin mRNA remained essentially unchanged. It is concluded that the differentiation of granulosa cells on ECM in vitro is associated with changes in cell shape and cell contacts and that such changes in the organization and expression of cytoskeletal proteins that are involved in determining these cellular structures.