Identification of differentially expressed microRNAs and their target genes in the hippocampal tissues of Fmr1 knockout mice.

Fragile X syndrome (FXS) is one of the most common forms of inherited mental retardation; it is usually associated with the transcriptional silencing of the Fmr1 gene and loss of its encoded protein, the fragile X mental retardation protein (FMRP). FMRP is an RNA-binding protein and participates in regulating the development of dendritic spines and synaptic plasticity. To uncover the possible role of microRNAs (miRNAs) in FXS and their relationship with FMRP, we used microarray analysis to investigate the miRNA expression profiles in the hippocampal tissues of Fmr1 knockout (Fmr1-KO) mice and wild type (WT) mice. A total of 75 differentially expressed miRNAs were identified, of which 58 were significantly upregulated and no miRNAs were significantly downregulated in Fmr1-KO mice. Quantitative real-time PCR (qRT-PCR) analysis was applied to validate the expression of 7 upregulated miRNAs; results indicated that the levels of only miR-449a and miR-720 were significantly upregulated. We further used bioinformatics software and databases to predict the target genes of these two miRNAs. The genes were related to dendritic spine development and synaptic plasticity; the qRT-PCR and western blotting results showed that cyclin-dependent kinase 5 (CDK5) and synaptotagmin 1 (SYT1) were differentially expressed in the Fmr1-KO mice and WT mice. In conclusion, this study evidenced diverse changes in the expression of miRNAs, and validated the miRNAs and their targeted genes in Fmr1-KO mice. Although further studies are required to better understand the function of miRNAs in FXS, the present research highlights a potential role of miRNAs in the pathogenesis of FXS. AJTR