Kiebler M A, Hemraj I, Verkade P, Köhrmann M, Fortes P, Marión R M, Ortín J, Dotti C G
European Molecular Biology Laboratory, 69012 Heidelberg, Germany.
J Neurosci. 1999 Jan 1;19(1):288-97. doi: 10.1523/JNEUROSCI.19-01-00288.1999.
In hippocampal neurons, certain mRNAs have been found in dendrites (), and their localization and translation have been implicated in synaptic plasticity (). One attractive candidate to achieve transport of mRNAs into dendrites is Staufen (Stau), a double-stranded RNA-binding protein, which plays a pivotal role in mRNA transport, localization, and translation in Drosophila (). Using antibodies raised against a peptide located in the RNA-binding domain IIa and a polyclonal antibody raised against a recently cloned human Staufen homolog, we identify a 65 kDa rat homolog in cultured rat hippocampal neurons. In agreement with the exclusive somatodendritic localization of mRNAs in these cells, we find that Staufen is restricted to the same domain. By immunoelectron microscopy, we show enrichment of the mammalian homolog of Stau (mStau) in the vicinity of smooth endoplasmic reticulum and microtubules near synaptic contacts. Finally, the association of the mStau with neuronal mRNAs is suggested by the colocalization with ribonucleoprotein particles specifically in distal dendrites known to contain mRNA, ribosomes, and translation factors (). These results suggest a role for mStau in the polarized transport and localization of mRNAs in mammalian neurons.
在海马神经元中,已在树突中发现某些信使核糖核酸(mRNA),其定位和翻译与突触可塑性有关。实现mRNA向树突运输的一个有吸引力的候选分子是Staufen(Stau),一种双链RNA结合蛋白,它在果蝇的mRNA运输、定位和翻译中起关键作用。利用针对位于RNA结合结构域IIa的一种肽产生的抗体以及针对最近克隆的人类Staufen同源物产生的多克隆抗体,我们在培养的大鼠海马神经元中鉴定出一种65 kDa的大鼠同源物。与这些细胞中mRNA仅存在于胞体-树突区域的定位一致,我们发现Staufen也局限于同一区域。通过免疫电子显微镜,我们显示Stau的哺乳动物同源物(mStau)在突触接触附近的光滑内质网和微管附近富集。最后,mStau与神经元mRNA的关联通过其与核糖核蛋白颗粒的共定位得以体现,特别是在已知含有mRNA、核糖体和翻译因子的远端树突中。这些结果表明mStau在哺乳动物神经元mRNA的极化运输和定位中发挥作用。