Institute of Neuroscience and Anatomy, Zhejiang University School of Medicine, Hangzhou, China.
Institute of Life Science, Zhejiang University, Hangzhou, China.
Gene. 2014 Sep 1;547(2):319-28. doi: 10.1016/j.gene.2014.06.063. Epub 2014 Jun 30.
Semaphorin3A (sema3a), mainly localized in the olfactory neuron layer and periglomerular layer, is essential for the normal arrangement of axons in the olfactory bulb both in embryonic and adult mice functioning through its dynamic spatiotemporal expression. The regulators that can modulate the expression of sema3a by direct interaction, however, are unknown. In order to find the regulators of sema3a in the olfactory bulb, we focused on microRNAs, well-known post-transcriptional regulators. We found that axon guidance is the main molecular and biological process ongoing in the steady-state olfactory bulb of the adult mouse by screening the abundant microRNAs and exploring their functions in the olfactory bulb via our customized microRNA arrays, Gene Ontology and Kyoto Encyclopedia of Genes annotation. Furthermore, we traced the expression of three candidate regulators (miR-30c, miR-200b, and miR-429) and sema3a by the quantitative real-time polymerase chain reaction and immunohistochemistry. The results showed that only miR-30c expression corresponded inversely with sema3a. Finally, miR-30c was verified to be a specific regulator of sema3a by dual luciferase reporter assay in vitro. Taken together, our results suggested that miR-30c is a potential regulator in axon-guidance by suppressing the expression of sema3a, which will give new insights in elucidating the mechanism of architectonic and functional maintenance of the olfactory bulb.
信号蛋白 3A(sema3a)主要定位于嗅神经元层和肾小球旁层,对于胚胎和成年期小鼠嗅球中轴突的正常排列是必不可少的,其通过动态的时空表达发挥作用。然而,能够通过直接相互作用调节 sema3a 表达的调节剂尚不清楚。为了在嗅球中找到 sema3a 的调节剂,我们专注于 microRNA,这是一种众所周知的转录后调节剂。我们通过筛选丰富的 microRNA 并通过我们定制的 microRNA 阵列、基因本体论和京都基因与基因组百科全书注释来探索它们在嗅球中的功能,发现轴突导向是成年小鼠嗅球中稳态下进行的主要分子和生物学过程。此外,我们通过定量实时聚合酶链反应和免疫组织化学追踪了三个候选调节剂(miR-30c、miR-200b 和 miR-429)和 sema3a 的表达。结果表明,只有 miR-30c 的表达与 sema3a 呈负相关。最后,通过体外双荧光素酶报告基因检测证实 miR-30c 是 sema3a 的特异性调节剂。总之,我们的结果表明,miR-30c 通过抑制 sema3a 的表达,是轴突导向的潜在调节剂,这将为阐明嗅球结构和功能维持的机制提供新的见解。