Regenerated leg segment patterns are regulated epigenetically by histone H3K27 methylation in the cricket Gryllus bimaculatus

Abstract

Hemimetabolous insects such as the cricket Gryllus bimaculatus regenerate lost tissue parts using blastemal cells, which is a population of dedifferentiated-proliferating cells. The gene expression of several epigenetic factors is upregulated in the blastema compared with the expression in differentiated tissue, suggesting that epigenetic changes in gene expression may control the differentiation status of blastema cells during regeneration. To clarify the molecular basis of epigenetic regulation during regeneration, we focused on the function of the Gryllus Enhancer of zeste (Gb’E(z)) and Ubiquitously-transcribed tetratricopeptide repeat gene on the X chromosome (Gb’Utx) homologues that regulate the methylation and demethylation on histone H3 27th lysine residue (H3K27), respectively. Methylated histone H3K27 in the regenerating leg was diminished by Gb’E(z)RNAi and was increased by Gb’UtxRNAi. Regenerated Gb’E(z)RNAi cricket legs exhibited extra leg segment formation between the tibia and tarsus, and regenerated Gb’UtxRNAi cricket legs showed leg joint formation defects in the tarsus. In the Gb’E(z)RNAi-regenerating leg, the Gb’dac expression domain expanded in the tarsus. In contrast, in the Gb’UtxRNAi-regenerating leg, Gb’Egfr expression in the middle of the tarsus was diminished. These results suggest that regulation of the histone H3K27 methylation state is involved in the repatterning process during leg regeneration among cricket species via the epigenetic regulation of leg patterning gene expression.