Dynes Joseph L, Steward Oswald
Reeve-Irvine Research Center, University of California, Irvine, Irvine, California 92697, USA.
J Comp Neurol. 2007 Jan 20;500(3):433-47. doi: 10.1002/cne.21189.
The mRNA for Arc (activity-regulated cytoskeletal protein) is delivered into dendrites and localizes selectively at active synapses. Here we use a green fluorescent protein-based labeling system and confocal microscopy to define the transport kinetics of exogenously expressed mRNA from chimaeric Arc constructs (Arc/MS2 mRNA) in the dendrites of living rat neurons in culture. Arc/MS2 mRNA assembles into particles that move independently, bidirectionally, and intermittently in a fashion indicative of transport. Transport velocities range from below 6 to 65 mum/minute, which is consistent with actin-based and microtubule-based transport, respectively. In general, orthograde translocations are longer than retrograde translocations. Rapidly translocating Arc/MS2 mRNA particles sometimes reverse direction and decrease velocity just before stopping, suggesting that local signals regulate Arc mRNA targeting movements. These observations identify several phases of Arc mRNA movement that serve as potential points for regulating Arc mRNA localization.
Arc(活性调节细胞骨架蛋白)的信使核糖核酸(mRNA)被输送到树突中,并选择性地定位于活跃突触处。在此,我们使用基于绿色荧光蛋白的标记系统和共聚焦显微镜,来确定培养的活大鼠神经元树突中外源表达的嵌合Arc构建体(Arc/MS2 mRNA)的mRNA转运动力学。Arc/MS2 mRNA组装成颗粒,以一种表明转运的方式独立、双向且间歇性地移动。转运速度范围从低于6微米/分钟到65微米/分钟,分别与基于肌动蛋白和基于微管的转运一致。一般来说,顺行易位比逆行易位更长。快速转运的Arc/MS2 mRNA颗粒有时会在停止前改变方向并降低速度,这表明局部信号调节Arc mRNA的靶向运动。这些观察结果确定了Arc mRNA运动的几个阶段,这些阶段可作为调节Arc mRNA定位的潜在关键点。