Rodionov Vladimir, Yi Julie, Kashina Anna, Oladipo Abiola, Gross Steven P
University of Connecticut Health Center, Department of Physiology and Center for Biomedical Imaging Technology, Farmington 06032-1507, USA.
Curr Biol. 2003 Oct 28;13(21):1837-47. doi: 10.1016/j.cub.2003.10.027.
Intracellular transport involves the movement of organelles along microtubules (MTs) or actin filaments (AFs) by means of opposite-polarity MT motors or actin-dependent motors of the myosin family. The correct delivery of organelles to their different destinations involves a precise coordination of the two transport systems. Such coordination could occur through regulation of the densities of the two cytoskeletal systems or through regulation of the activities of the cytoskeletal motors by signaling mechanisms.
To investigate the mechanisms of switching between MT and AF-dependent transport, we examine the influence of the densities of the MT and AF network on pigment transport in fish melanophores. We also change signaling by using activators and inhibitors of Protein Kinase A (PKA). We find that the key parameters characterizing pigment granule transport along MTs do not depend on MT density and are not significantly altered by complete disruption of AFs. In contrast, the kinetics of changes in these parameters correlate with the kinetics of changes in the intracellular levels of cAMP and are affected by the inhibitors of PKA, suggesting the regulation of MT- and AF-dependent motors by cAMP-induced signaling. Furthermore, perturbation of cAMP levels prevents the transfer of pigment granules from MTs onto AFs.
We conclude that the switching of pigment granules between the two major cytoskeletal systems is independent of the densities of MT or AF but is tightly controlled by signaling events.
细胞内运输涉及细胞器借助微管(MTs)或肌动蛋白丝(AFs),通过相反极性的MT马达或肌球蛋白家族的肌动蛋白依赖性马达进行移动。将细胞器正确运送到其不同目的地需要两个运输系统的精确协调。这种协调可以通过调节两个细胞骨架系统的密度或通过信号机制调节细胞骨架马达的活性来实现。
为了研究MT和AF依赖性运输之间的转换机制,我们研究了MT和AF网络密度对鱼类黑素细胞中色素运输的影响。我们还通过使用蛋白激酶A(PKA)的激活剂和抑制剂来改变信号传导。我们发现,表征色素颗粒沿MTs运输的关键参数不依赖于MT密度,并且在AFs完全破坏时不会显著改变。相反,这些参数变化的动力学与细胞内cAMP水平变化的动力学相关,并受PKA抑制剂的影响,这表明cAMP诱导的信号传导对MT和AF依赖性马达有调节作用。此外,cAMP水平的扰动会阻止色素颗粒从MTs转移到AFs上。
我们得出结论,色素颗粒在两个主要细胞骨架系统之间的转换与MT或AF的密度无关,但受信号事件的严格控制。