MicroRNAs regulate synthesis of the neurotransmitter substance P in human mesenchymal stem cell-derived neuronal cells
MicroRNAs regulate synthesis of the neurotransmitter substance P in human mesenchymal stem cell-derived neuronal cells
复制标题
DOI：
10.1073/pnas.0703037104
复制
发表时间：
2007-09-25
影响因子：
11.1
通讯作者：
Rameshwar, Pranela
中科院分区：
文献类型：
--
作者：
Greco, Steven J.;Rameshwar, Pranela
MicroRNAs （miRNAs）是一类19- 23nt的非编码小rna，可结合靶mrna的3' UTR介导动物的翻译抑制。mirna已被证明可以调节发育过程，如干细胞的自我更新、神经元分化、肌肉发生和癌症。mirna在调节神经递质合成中的功能作用尚未确定。我们以间充质干细胞（MSCs）为模型，研究mirna介导的神经递质调控在发育中的神经细胞。间充质干细胞是中胚层来源的细胞，主要存在于成人骨髓中，可以产生功能性神经细胞。我们之前已经证明，人类间质干细胞衍生的神经细胞表达神经递质基因Tac1，但除非受到炎症介质IL-1 α的刺激，否则不会合成该基因编码的肽，即神经递质物质P （SP）。这些发现提示了mirna在SP合成调控中的潜在作用。在这里，我们使用miRNA特异性生物阵列报道了未分化的人间质干细胞和间质干细胞衍生的神经元细胞的miRNA谱。通过miRanda算法分析IL-1 α刺激后神经元细胞中增加和减少的mirna，以预测Tac1 mRNA靶标。推测的miR-130a、miR-206和miR-302a结合位点在Tac1的3' UTR内。使用荧光素酶报告系统进行靶标验证，确认了miR130a和miR-206位点。神经元细胞中miR-130a和miR-206的特异性抑制导致SP的合成和释放。这些研究为ml在调节神经递质合成中的新调控作用提供了一种不同的途径。
MicroRNAs (miRNAs) are a class of 19- to 23-nt, small, noncoding RNAs, which bind the 3' UTR of target mRNAs to mediate translational repression in animals. miRNAs have been shown to regulate developmental processes, such as self-renewal of stem cells, neuronal differentiation, myogenesis, and cancer. A functional role of miRNAs in the regulation of neurotransmitter synthesis has yet to be ascribed. We used mesenchymal stem cells (MSCs) as a model to study miRNA-mediated neurotransmitter regulation in developing neuronal cells. MSCs are mesoderm-derived cells, primarily resident in adult bone marrow, which can generate functional neuronal cells. We have previously shown that human MSC-derived neuronal cells express the neurotransmitter gene, Tac1, but do not synthesize the gene's encoded peptide, the neurotransmitter substance P (SP), unless stimulated with the inflammatory mediator IL-1 alpha. These findings suggested a potential role for miRNAs in the regulation of SP synthesis. Here, we report on the miRNA profile of undifferentiated human MSCs and MSC-derived neuronal cells by using miRNA-specific bioarrays. miRNAs that were increased in the neuronal cells and decreased after IL-1 alpha stimulation were analyzed by the miRanda algorithm to predict Tac1 mRNA targets. Putative miR-130a, miR-206, and miR-302a binding sites were predicted within the 3' UTR of Tac1. Target validation using a luciferase reporter system confirmed the miR130a and miR-206 sites. Specific inhibition of miR-130a and miR-206 in the neuronal cells resulted in SP synthesis and release. The studies provide a different approach in ascribing a new regulatory role for ml in regulating neurotransmitter synthesis.