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首页> 外文期刊>Molecular biology of the cell >An InCytes from MBC Selection: Microtubule-mediated Src Tyrosine Kinase Trafficking in Neuronal Growth Cones
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An InCytes from MBC Selection: Microtubule-mediated Src Tyrosine Kinase Trafficking in Neuronal Growth Cones

机译：从MBC选择的InCytes：神经元生长锥中的微管介导的Src酪氨酸激酶贩运。

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Src family tyrosine kinases are important signaling enzymes in the neuronal growth cone, and they have been implicated in axon guidance; however, the detailed localization, trafficking, and cellular functions of Src kinases in live growth cones are unclear. Here, we cloned two novel Aplysia Src kinases, termed Src1 and Src2, and we show their association with both the plasma membrane and the microtubule cytoskeleton in the growth cone by live cell imaging, immunocytochemistry, and cell fractionation. Activated Src2 is enriched in filopodia tips. Interestingly, Src2-enhanced green fluorescent protein–positive endocytic vesicles and tubulovesicular structures undergo microtubule-mediated movements that are bidirectional in the central domain and mainly retrograde in the peripheral domain. To further test the role of microtubules in Src trafficking in the growth cone, microtubules were depleted with either nocodazole or vinblastine treatment, resulting in an increase in Src2 plasma membrane levels in all growth cone domains. Our data suggest that microtubules regulate the steady-state level of active Src at the plasma membrane by mediating retrograde recycling of endocytosed Src. Expression of constitutively active Src2 results in longer filopodia that protrude from smaller growth cones, implicating Src2 in controlling the size of filopodia and lamellipodia.
机译：Src家族的酪氨酸激酶是神经元生长锥中重要的信号转导酶，它们已经参与了轴突的引导。但是，尚不清楚Src激酶在活生长锥中的详细定位，运输和细胞功能。在这里，我们克隆了两个新的Aplysia Src激酶，分别称为Src1和Src2，并通过活细胞成像，免疫细胞化学和细胞分级分离显示了它们与质膜和生长锥中的微管细胞骨架的相关性。激活的Src2富含丝状伪足提示。有趣的是，Src2增强的绿色荧光蛋白阳性的内吞囊泡和微管小泡结构经历了微管介导的运动，这些运动在中央区域是双向的，在周边区域主要是逆行的。为了进一步测试微管在生长锥中Src转运中的作用，用nocodazole或长春碱处理消除了微管，导致所有生长锥结构域中Src2质膜水平增加。我们的数据表明，微管通过介导内吞Src的逆行循环来调节质膜上活性Src的稳态水平。组成型活性Src2的表达导致更长的丝状伪足从较小的生长锥突出，这暗示Src2可以控制丝状伪足和片状脂质体的大小。

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