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A MicroRNA Profile in Fmr1 Knockout Mice Reveals MicroRNA Expression Alterations with Possible Roles in Fragile X Syndrome

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Abstract

Fragile X syndrome (FXS), a common form of inherited mental retardation, is caused by a loss of expression of the fragile X mental retardation protein (FMRP). FMRP is involved in brain functions by interacting with mRNAs and microRNAs (miRNAs) that selectively control gene expression at translational level. However, little is known about the role of FMRP in regulating miRNA expression. Here, we found a development-dependant dynamic expression of Fmr1 gene (encoding FMRP) in mouse hippocampus with a small peak at postnatal day 7 (P7). MiRNA microarray analysis showed that the levels of 38 miRNAs showed a significant increase with about 15 ~ 250-folds and the levels of 26 miRNAs showed a significant decrease with only about 2 ~ 4-folds in the hippocampus of P7 Fmr1 knockout (KO) mice. The qRT-PCR assay showed that nine of the most increased miRNAs (>100-folds in microarrays) increased about 40 ~ 70-folds and their pre-miRNAs increased about 5 ~ 10-folds, but no significant difference in their pri-miRNA levels was observed, suggesting that the alterations of partial miRNAs are an indirect consequence of FMRP lacking. We further demonstrated that a set of protein-coding mRNAs, potentially targeted by the nine miRNAs, were down-regulated in the hippocampus of Fmr1 KO mice. Finally, luciferase assays demonstrated that miR-34b, miR-340, and miR-148a could down-regulate the reporter gene expression by interacting with the Met 3′ UTR. Taken together, these findings suggest that the miRNA expression alterations resulted from the absence of FMRP might contribute to molecular pathology of FXS.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (grant numbers 81171073, 30870876, 81371436, 31070928, and 81271434), the Guangzhou Scholar Project (grant number 10A011G), and the Scientific Research of Guangzhou Municipal Colleges and Universities (grant number 10A211). We are indebted to Dr. Ben A. Oostra (Erasmus University, Netherland) for kindly providing the FVB Fmr1 knockout mice and the FVB wild-type mice. We are grateful to the He Shanheng Charity Foundation for contributing to the development of this institute.

Conflict of Interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Key Laboratory of Neurogenetics and Channelopathies of Guangdong Province and the Ministry of Education of China, Collaborative Innovation Center for Neurogenetics and Channelopathies, Institute of Neuroscience and the Second Affiliated Hospital of Guangzhou Medical University, 250 Changgang East Road, Guangzhou, 510260, China

    Ting Liu, Rui-Ping Wan, Ling-Jia Tang, Shu-Jing Liu, Hai-Jun Li, Qi-Hua Zhao, Wei-Ping Liao, Yong-Hong Yi & Yue-Sheng Long

  2. Key Laboratory for Major Obstetric Diseases of Guangdong Province and Key laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, Guangzhou Medical University, Guangzhou, China

    Hai-Jun Li & Xiao-Fang Sun

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  1. Ting Liu
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Correspondence to Yong-Hong Yi or Yue-Sheng Long.

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Ting Liu and Rui-Ping Wan contributed equally to this work.

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Fig. S1

The mRNAs of partial miRNA target genes present in the mouse hippocampus. These data of the mRNA levels determined by in situ hybridization were downloaded from the public data from Allen Brain Atlas Database (http://www.brain-map.org/) (GIF 133 kb)

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Liu, T., Wan, RP., Tang, LJ. et al. A MicroRNA Profile in Fmr1 Knockout Mice Reveals MicroRNA Expression Alterations with Possible Roles in Fragile X Syndrome. Mol Neurobiol 51, 1053–1063 (2015). https://doi.org/10.1007/s12035-014-8770-1

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