Comprehensive Analysis of Differences of N6-Methyladenosine of Lncrnas Between Atrazine-Inducedand Normal Xenopus Laevis Testis

Abstract Background: Increasing evidence suggested N6-methyladenosine (m6A) plays an important role in RNA stability, degradation, splicing and translation. M6A is found in different RNA including long non-coding RNA (lncRNA) which has been found possess significant biological functions. Our previous study examined the m6A profile of mRNAs in testis tissues of Xenopus laevis (X. laevis) with and without treatment with 100 µg/L atrazine (AZ). The result revealed that m6A is a highly conserved modification across the species. Methods: In this study, we apply previous approach to further investigate m6A modification profile of lncRNAs and predict the potential mechanism. In brief, m6A was sequenced by MeRIP sequencing using the latest Illumina HiSeq sequencer. Pathway enrichment analysis was used to maps genes to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Results: The results showed that m6A of lncRNAs enriched around intergenic region in testes of X. laevis. We further investigated the differential expression of lncRNAs m6A in testes of AZ-exposed compared with that in animals from control group. The results indicated that up to 198 differentially methylated m6A sites were detected within 188 lncRNAs, in which 89 sites were significantly up-regulated and 109 sites were significantly down-regulated. Data from Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that AZ-affected lncRNAs m6A sites were mainly involved in 10 pathways in which 3 mutual pathways were found in the result of differentially m6A-methylated mRNAs. Conclusions: These findings suggest that differentially m6A-methylated lncRNAs and these 3 pathways may act on regulatory roles in abnormal testis development of AZ-exposed X. laevis. This study for the first time provide insights into the profile of lncRNAs m6A modifications in amphibian species.