Comparison And Analysis Of MiRNAs Involved In The Regulation Under Microgravity In Caenorhabditid Elegans

MiRNAs are a class of non-coding RNAs which participate in the growth and development, locomotion and oviposition by regulating the expression level of the targeted genes. Our previous study performed miRNA microarray analysis in the wild-type (WT) and dys-1 mutant strains of dauer larvae after Shenzhou-8 spacecraft mission and found that cel-miR-52,56,124 may involve in regulating the locomotion-and behavior under microgravity condition. Therefore, this study is to investigate whether cel-miR-52,56,124 participated in locomotion and oviposition of WT and dys-1 mutant under the microgravity environment.Our research observed and proved that the dys-1 mutant worms owned the head over-bending. The locomotion and oviposition were similar between WT and dys-1 mutant worms. qRT-PCR was further used to determine and found the expression levels of miR-52 and 56 were significantly increased at dauer stage of dys-1 mutant but not different at the adult stage, indicading that miR-52,56 may play a biological role in dys-1 mutant dauer. However, the expression level of miR-124 showed significant changes at both dauer and adult stage, which indicated that miR-124 may be related to the loss of dys-1 but not the development period.To determine the locomotion, oviposition and expression levels of the three miRNAs and the target genes of the nematode FO and F1 generation, we used the 3D clinostat to simulate microgravity for 16.5 days. The results indicated that locomotion ability was reduced for FO generation in both WT and dys-1 mutation worms, but the WT was by significant reduced compared to WT by simulating microgravity. For the F1 generation, the reduction of locomotion was observed only in WT worms under microgravity condition, implying that the dys-1 mutation was insensitive to microgravity. miR-52,56 and miR-124 of dys-1 mutant were expressed differently by simulating microgravity. The target genes of nep-11 and C07H4.1 of miR-52 and 56 may involve in regulating the reduction of locomotion in both WT and dys-1 mutation. C07H4.1 may relate to the locomation of F1 generation WT and dys-1 mutation under microgravity condition, and thus the gene was speculated to participate in the functional regulation of neuromuscular system. The amounts of spawning were no significant different at different developmentl period in both WT and dys-1 mutant worms.Our research illustrated the expression levels of three miRNAs (cel-miR-52,56 and 124) at the dauer and adult stage in WT and mutant worms. The three miRNAs were speculated to involve in the regulation of locomotion of microgravity-treated mutant worms by analyzing the expression of miRNAs and their target genes. The result provid an important evidence that the dys-1 gene involved in the molecular machanism of muscle atrophy under microgravity condition.