Somatodendritic microRNAs identified by laser capture and multiplex RT-PCR
Somatodendritic microRNAs identified by laser capture and multiplex RT-PCR
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DOI：
10.1261/rna.480407
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发表时间：
2007-08-01
期刊：
影响因子：
4.5
通讯作者：
Kosik, Kenneth S.
中科院分区：
文献类型：
--
作者：
Kye, Min-Jeong;Liu, Tsunglin;Kosik, Kenneth S.
树突中存在的RNA目录代表了有助于突触可塑性的局部翻译的完整库。大多数观点认为，树突状mRNA库被选择性地转运到树突状目的地。这一观点要求树突中的一些mRNA相对于细胞体局部富集;然而，支持这一假设的定量比较目前还不存在。这些与mRNAs的体树突分布相关的问题也适用于microRNAs，类似于21个核苷酸的非编码转录本，它们与靶mRNA结合，抑制其翻译或使其不稳定。我们将激光捕获与多重真实的-时间RT（逆转录）PCR相结合，分别定量神经炎和体细胞隔室中的microRNA。通过RT-PCR测量这两个隔室中的一组mRNA（包括已知的树突状mRNA）来标准化样品。大多数神经元miRNA在树突中检测到。除了少数显著的例外，大多数miRNA通过体树突隔室以几乎恒定的梯度分布。因此，对于丰度较低的miRNA，miRNA的总神经元浓度可以在细胞体中保持容易检测，但从树突中消失。非常少数量的miRNA偏离整个miRNA群体的分布梯度，因为在树突中相对富集或耗尽。
The catalog of RNAs present in dendrites represents the complete repertoire of local translation that contributes to synaptic plasticity. Most views hold that a pool of dendritic mRNAs is selectively transported to a dendritic destination. This view requires that some mRNAs in the dendrite are locally enriched relative to the cell body; however, quantitative comparisons that would support this assumption do not currently exist. These issues related to somatodendritic distribution of mRNAs also apply to the microRNAs, similar to 21 nucleotide noncoding transcripts that bind to target mRNAs and either inhibit their translation or destabilize them. We combined laser capture with multiplex real- time RT (reverse transcription) PCR to quantify microRNAs in the neuritic and somatic compartments separately. The samples were standardized by RT-PCR measurements of a set of mRNAs, including known dendritic mRNAs, in these two compartments. Most neuronal miRNAs were detected in dendrites. With a few notable exceptions, most miRNAs were distributed through the somatodendritic compartment across a nearly constant gradient. Thus for lower-abundance miRNAs, the total neuronal concentration of the miRNA can remain readily detectable in the cell body but vanish from the dendrite. A very small number of miRNAs deviate from the distribution gradient across the miRNA population as relatively enriched or depleted in the dendrite.