Differential participation of actin-and tubulin-based vesicle transport systems during secretion in bovine chromaffin cells
Differential participation of actin-and tubulin-based vesicle transport systems during secretion in bovine chromaffin cells
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DOI：
10.1046/j.1460-9568.2003.02801.x
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发表时间：
2003-08-01
影响因子：
3.4
通讯作者：
Gutiérrez, LM
中科院分区：
文献类型：
--
作者：
Neco, P;Giner, D;Gutiérrez, LM
在培养的牛肾上腺髓质嗜铬细胞中，研究了胞吐过程中细胞骨架元件在囊泡运输中的作用。电流测定的去极化依赖性的儿茶酚胺释放与肌动蛋白或肌球蛋白抑制剂处理的单个完整的细胞显示，与未处理的细胞相比，在分泌的快，慢相的变化。相比之下，微管分解剂或稳定剂对分泌有中等影响，仅影响缓慢分泌组分的释放。在使用共聚焦动态显微镜的实验中，我们已经观察到肌动蛋白和肌球蛋白抑制剂在废除整个细胞质中的囊泡运动的剧烈作用，以及微管稳定剂引起的深核周区域中的颗粒流动性的抑制。失去流动性后，囊泡与F-肌动蛋白或微管的细丝相关联。此外，皮层囊泡的流动性受到肌动蛋白-肌球蛋白抑制剂的影响，但不受微管抑制剂。对活细胞中皮质细胞骨架的研究表明，囊泡与致密的管状F-肌动蛋白结构相关，微管呈低密度网络状。这些研究结果表明，在皮质区的细胞骨架元素的分布和密度可能会影响在分泌囊泡池的招聘。
The role of cytoskeletal elements in vesicle transport occurring during exocytosis was examined in adrenal medullary bovine chromaffin cells maintained in culture. Amperometric determination of depolarization-dependent catecholamine release from individual intact cells treated with actin or myosin inhibitors showed alterations in the fast and slow phases of secretion when compared with untreated cells. In contrast, microtubule disassemblers or stabilizers have a moderate effect on secretion, only affecting the release of slow secretory components. In experiments using confocal dynamic microscopy we have observed the drastic effect of actin and myosin inhibitors in abolishing vesicle movement throughout the cytoplasm, and the inhibition of granule mobility in deep perinuclear regions caused by the microtubule stabilizers. Following loss of mobility, vesicles were associated with filaments of F-actin or microtubules. In addition, the mobility of cortical vesicles was affected by actin-myosin inhibitors but not by microtubule inhibitors. The study of cortical cytoskeleton in living cells showed vesicles associated with dense tubular F-actin structures, with microtubules appearing as low density networks. These findings suggest that the distribution and density of both cytoskeletal elements in the cortical region may influence the recruitment of vesicle pools during secretion.