Visualization of peroxisomes in living plant cells reveals acto-myosin-dependent cytoplasmic streaming and peroxisome budding
Visualization of peroxisomes in living plant cells reveals acto-myosin-dependent cytoplasmic streaming and peroxisome budding
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DOI：
10.1093/pcp/pcf045
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发表时间：
2002-04-01
影响因子：
4.9
通讯作者：
Chua, NH
中科院分区：
文献类型：
--
作者：
Jedd, G;Chua, NH
在这里，我们使用表达绿色荧光蛋白(GFP)和过氧化物酶体靶向信号1(PTS1)的转基因拟南芥植物来检测活植物细胞中的过氧化物体。利用延时激光扫描共聚焦显微镜，我们发现植物过氧化物体呈现快速的定向运动，峰值速度接近10微米S(-1)。与哺乳动物的过氧化体在微管上运动不同，植物的过氧化物酶体的运动依赖于肌动蛋白微丝和肌球蛋白马达，因为它分别被菌丝素B和丁二酮单肟处理所阻断。相比之下，破坏微管的药物对过氧化物酶体流没有影响。通过分别使用GFP-talin和GFP-PTS1融合蛋白同时显示活细胞中的肌动蛋白细丝和过氧酶体，进一步证明了过氧酶体与肌动蛋白细胞骨架的联系。此外，还观察到了过氧化体的萌发，这可能是植物过氧化体增殖的一种机制。与GFP-PTS1标记相关的强信号也使我们能够观察不同类型细胞中的细胞质流动。在细长的细胞和参与长距离运输的细胞中，过氧化物体的运动最强烈，这表明高等植物利用细胞质流动来帮助小泡和细胞器进行长距离的运输。
Here we examine peroxisomes in living plant cells using transgenic Arabidopsis thaliana plants expressing the green fluorescent protein (GFP) fused to the peroxisome targeting signal 1 (PTS1). Using time-lapse laser scanning confocal microscopy we find that plant peroxisomes exhibit fast directional movement with peak velocities approaching 10 mum s(-1). Unlike mammalian peroxisomes which move on microtubules, plant peroxisome movement is dependent on actin microfilaments and myosin motors, since it is blocked by treatment with latrunculin B and butanedione monoxime, respectively. In contrast, microtubule-disrupting drugs have no effect on peroxisome streaming. Peroxisomes were further shown to associate with the actin cytoskeleton by the simultaneous visualization of actin filaments and peroxisomes in living cells using GFP-talin and GFP-PTS1 fusion proteins, respectively. In addition, peroxisome budding was observed, suggesting a possible mechanism of plant peroxisome proliferation. The strong signal associated with the GFP-PTS1 marker also allowed us to survey cytoplasmic streaming in different cell types. Peroxisome movement is most intense in elongated cells and those involved in long distance transport, suggesting that higher plants use cytoplasmic streaming to help transport vesicles and organelles over long distances.