Cell density‐dependent decrease in cytoskeletal actin and myosin in cultured osteoblastic cells: Correlation with cyclic AMP changes
Cell density‐dependent decrease in cytoskeletal actin and myosin in cultured osteoblastic cells: Correlation with cyclic AMP changes
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培养的成骨细胞中细胞骨架肌动蛋白和肌球蛋白的细胞密度依赖性减少：与环 AMP 变化的相关性
DOI：
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发表时间：
1991
影响因子：
4
通讯作者：
G. Rodan
中科院分区：
文献类型：
--
作者：
J. Egan;G. Gronowicz;G. Rodan
在骨发育过程中，成骨细胞沿着最近沉积的类骨质形成连续层，其形态从成纤维细胞样变为立方形。在培养中，随着细胞密度的增加，也会发生类似的变化。我们研究了环AMP在这一过程中可能发挥的作用，因为据报道，环AMP在成纤维细胞中随着细胞密度的增加而增加，并且在对甲状旁腺激素的反应中观察到类似的形状变化，甲状旁腺激素也增加了成骨细胞中的细胞环AMP。以增加的密度接种成骨细胞富集的大鼠颅骨细胞。通过聚丙烯酰胺凝胶电泳估计Triton X-100可提取和不可提取的肌动蛋白和肌球蛋白之间的分布。通过用3H-腺嘌呤预标记细胞ATP池，然后通过高效液相色谱法提取和分离3H-cAMP来估计细胞内cAMP。我们发现成骨细胞每个细胞含有约40 pg肌动蛋白和5.3 pg肌球蛋白。在细胞密度为10，000至50，000个细胞/cm 2时，约60%的肌动蛋白和70%的肌球蛋白处于不可提取（交联）形式。高于50，000个细胞/cm 2时，交联肌动蛋白和肌球蛋白呈细胞密度依赖性减少，同时细胞环AMP增加。与磷酸二酯酶抑制剂孵育产生的环AMP的可比上升，以及与其他增加环AMP的药物治疗产生了类似的细胞骨架肌动蛋白和肌球蛋白水平的下降。细胞松弛素B治疗，通过其对肌动蛋白聚合的影响，产生类似的细胞形状和细胞骨架肌动蛋白的变化。研究结果表明，细胞内环AMP的升高可能在成骨细胞中观察到的细胞形状和微丝组织的密度依赖性变化中发挥作用。
During bone development, osteoblasts form a contiguous layer along recently deposited osteoid and their morphology changes from fibroblast‐like to cuboidal. In culture, similar changes occur with increased cell density. We examined the possible role of cyclic AMP in this process since cyclic AMP was reported to increase in fibroblasts with increased cell density and similar shape changes were seen in response to parathyroid hormone, which also increases cellular cyclic AMP in osteoblastic cells. Osteoblast‐enriched rat calvaria cells were seeded at increasing density. The distribution between Triton X‐100 extractable and nonextractable actin and myosin was estimated by polyacrylamide gel electrophoresis. Intracellular cyclic AMP was estimated by prelabeling the cellular ATP pool with 3H‐adenine, followed by extraction and separation of 3H‐cAMP by high‐performance liquid chromatography. We found that osteoblastic cells contain about 40 pg actin and 5.3 pg myosin per cell. Around 60% of the actin and 70% of the myosin were in the nonextractable (crosslinked) form at cell densities of 10,000 to 50,000 cells per cm2. Above 50,000 cells/cm2, there was a cell density‐dependent reduction in crosslinked actin and myosin and a concomitant increase in cellular cyclic AMP. A comparable rise in cyclic AMP, produced by incubation with phosphodiesterase inhibitors, and treatment with other agents that increase cyclic AMP produced a similar decrease in the level of cytoskeletal actin and myosin. Cytochalasin B treatment, through its effect on actin polymerization, produced similar changes in cell shape and cytoskeletal actin. The findings suggest that an elevation in intracellular cyclic AMP may play a role in the density‐dependent changes in cell shape and microfilament organization observed in osteoblasts.
兔中性粒细胞中与细胞骨架相关的肌动蛋白趋化因子的刺激。
DOI：
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发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
White,JR;Naccache,PH;Sha'afi,RI
通讯作者：
Sha'afi,RI
凝血酶激活后人血小板细胞骨架结构的变化。
DOI：
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发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Jennings,LK;Fox,JE;Edwards,HH;Phillips,DR
通讯作者：
Phillips,DR