Mayford M, Baranes D, Podsypanina K, Kandel E R
Center for Neurobiology and Behavior College of Physicians and Surgeons, Columbia University, New York, NY, USA.
Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13250-5. doi: 10.1073/pnas.93.23.13250.
Neuronal signaling requires that synaptic proteins be appropriately localized within the cell and regulated there. In mammalian neurons, polyribosomes are found not just in the cell body, but also in dendrites where they are concentrated within or beneath the dendritic spine. The alpha subunit of Ca(2+)-calmodulin-dependent protein kinase II (CaMKII alpha) is one of only five mRNAs known to be present within the dendrites, as well as in the soma of neurons. This targeted subcellular localization of the mRNA for CaMKII alpha provides a possible cell biological mechanism both for controlling the distribution of the cognate protein and for regulating independently the level of protein expression in individual dendritic spines. To characterize the cis-acting elements involved in the localization of dendritic mRNA we have produced two lines of transgenic mice in which the CaMKII alpha promoter is used to drive the expression of a lacZ transcript, which either contains or lacks the 3'-untranslated region of the CaMKII alpha gene. Although both lines of mice show expression in forebrain neurons that parallels the expression of the endogenous CaMKII alpha gene, only the lacZ transcripts bearing the 3'-untranslated region are localized to dendrites. The beta-galactosidase protein shows a variable level of expression along the dendritic shaft and within dendritic spines, which suggests that neurons can control the local biochemistry of the dendrite either through differential localization of the mRNA or variations in the translational efficiency at different sites along the dendrite.
神经元信号传导要求突触蛋白在细胞内正确定位并在那里受到调节。在哺乳动物神经元中,多核糖体不仅存在于细胞体中,也存在于树突中,它们集中在树突棘内或其下方。钙调蛋白依赖性蛋白激酶II(CaMKIIα)的α亚基是已知仅有的五种在树突以及神经元胞体中存在的mRNA之一。CaMKIIα mRNA的这种靶向亚细胞定位为控制同源蛋白的分布以及独立调节单个树突棘中蛋白表达水平提供了一种可能的细胞生物学机制。为了表征参与树突mRNA定位的顺式作用元件,我们培育了两系转基因小鼠,其中CaMKIIα启动子用于驱动lacZ转录本的表达,该转录本要么包含要么缺少CaMKIIα基因的3'非翻译区。尽管两系小鼠在前脑神经元中的表达都与内源性CaMKIIα基因的表达平行,但只有带有3'非翻译区的lacZ转录本定位于树突。β-半乳糖苷酶蛋白在树突轴和树突棘内显示出不同水平的表达,这表明神经元可以通过mRNA的差异定位或沿树突不同位点翻译效率的变化来控制树突的局部生物化学过程。