Huang Fen, Chotiner Jennifer K, Steward Oswald
Reeve-Irvine Research Center, Department of Anatomy and Neurobiology, University of California, Irvine, California 92697, USA.
J Neurosci. 2005 Aug 3;25(31):7199-209. doi: 10.1523/JNEUROSCI.1779-05.2005.
There is increasing evidence that long-lasting forms of activity-dependent synaptic plasticity, such as long-term potentiation (LTP) and long-term depression (LTD), require local synthesis of proteins within dendrites. Identifying novel dendritic mRNAs and determining how their distribution and translation is regulated is a high priority. We demonstrate here that the mRNA for the elongation factor 1 alpha (EF1alpha) is present in vivo in the dendrites of neurons that exhibit LTP and LTD, and that its translation is locally regulated. The subcellular distribution of EF1alpha mRNA differs from any of the dendritic mRNAs that have been described previously. In the hippocampus, the mRNA is highly expressed in cell bodies and is also concentrated in the zone of termination of commissural/associational afferents in the inner molecular layer, suggesting that mRNA localization is in some way related to the distribution of different types of synapses. Nevertheless, the localization of EF1alpha mRNA is not altered by prolonged periods of synaptic activation that are sufficient to cause a dramatic redistribution of Arc mRNA. Local application of the metabotropic glutamate receptor agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) led to dramatic increases in immunostaining for EF1alpha protein in dendrites, and treatment of hippocampal slices with DHPG, which is known to induce LTD, led to increases in EF1alpha protein levels. Both responses were blocked by the protein synthesis inhibitor anisomycin. In contrast, stimulation of the perforant path using patterns of stimulation that induce LTP caused rapid increases of immunostaining for EF1alpha protein in the activated dendritic lamina, but these increases were not blocked by anisomycin or rapamycin. The findings suggest that local synthesis of EF1alpha protein may be important for the synaptic mechanisms that underlie protein synthesis-dependent LTD.
越来越多的证据表明,长期存在的依赖活动的突触可塑性形式,如长时程增强(LTP)和长时程抑制(LTD),需要在树突内进行蛋白质的局部合成。识别新的树突mRNA并确定其分布和翻译如何受到调控是当务之急。我们在此证明,延伸因子1α(EF1α)的mRNA在体内存在于表现出LTP和LTD的神经元的树突中,并且其翻译受到局部调控。EF1α mRNA的亚细胞分布不同于先前描述的任何树突mRNA。在海马体中,该mRNA在细胞体中高度表达,并且还集中在内分子层中连合/联合传入纤维的终止区域,这表明mRNA定位在某种程度上与不同类型突触的分布有关。然而,EF1α mRNA的定位不会因足以导致Arc mRNA发生显著重新分布的长时间突触激活而改变。代谢型谷氨酸受体激动剂(R,S)-3,5-二羟基苯甘氨酸(DHPG)的局部应用导致树突中EF1α蛋白的免疫染色显著增加,并且用已知可诱导LTD的DHPG处理海马切片会导致EF1α蛋白水平升高。这两种反应均被蛋白质合成抑制剂茴香霉素阻断。相比之下,使用诱导LTP的刺激模式刺激穿通路径会导致活化树突层中EF1α蛋白的免疫染色迅速增加,但这些增加不受茴香霉素或雷帕霉素的阻断。这些发现表明,EF1α蛋白的局部合成可能对蛋白质合成依赖性LTD的突触机制很重要。