Comprehensive analysis of differences of N6-methyladenosine of lncRNAs between atrazine-induced and normal Xenopus laevis testis.

Background: Increasing evidence suggested N⁶-methyladenosine (m⁶A) modification is crucial for male germline development. However, m⁶A modification of lncRNAs gains a little attention in amphibians in recent years. Xenopus laevis (X. laevis) was chosen to be an ideal model organism for testing environmental endocrine disrupting chemicals (EDCs) exposure and resultant effects. Atrazine (AZ) as an endocrine disrupt can effect development of testis in amphibians. Our previous study revealed that m⁶A is a highly conserved modification across the species.

Results: The results of m⁶A sequences showed that m⁶A-methylated lncRNAs enriched in intergenic region in testes of X. laevis. We further examined the differential expression of lncRNAs m⁶A sites in testes of AZ-exposed and 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 significantly up-methylated sites and 109 significantly down-methylated sites. Data from 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 suggested 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 provides insights into the profile of lncRNAs m⁶A modifications in amphibian species.