Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological pro...Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological processes including development,aging,metabolism,and immunity,however the functions of KDM4A in fish are still unclear.There are two copies of the kdm4a gene in zebrafish,namely kdm4aa and kdm4ab,kdm4aa was edited using CRISPR/Cas9 technology in the present study,then homozygous kdm4aa mutants(kdm4aa^(-/-))were obtained,and loss of kdm4aa was confirmed by sequencing and increased H3K9me3.Whole-mount in situ hybridization showed that kdm4aa is widely expressed during the embryonic development of zebrafish.Compared with WT zebrafish,kdm4aa^(-/-)zebrafish showed no significant difference in gamete formation and fertilization,but the survival rate of kdm4aa^(-/-)embryos dramatically reduced to 21%at 26 hpf.Further observation showed that about 80%of survived kdm4aa^(-/-)zebrafish experienced disruption in stripe formation,and 10%of survived kdm4aa^(-/-)zebrafish underwent vertebral malformation.Alizarin red S staining demonstrated the abnormal spinal development in kdm4aa^(-/-)zebrafish.These results indicated that kdm4aa is required for normal embryonic development of zebrafish,loss of kdm4aa function leads to decreased survival during the early stages of zebrafish development and morphological variation in adult zebrafish.展开更多
基金from Shanghai Agriculture Applied Technology Development Program,China(Grant No.G20190102)National Natural Science Foundation of China(Grant No.81770165 to BH)Shanghai Municipal Education Commission(Project for Gaofeng Discipline of Fishery).
文摘Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological processes including development,aging,metabolism,and immunity,however the functions of KDM4A in fish are still unclear.There are two copies of the kdm4a gene in zebrafish,namely kdm4aa and kdm4ab,kdm4aa was edited using CRISPR/Cas9 technology in the present study,then homozygous kdm4aa mutants(kdm4aa^(-/-))were obtained,and loss of kdm4aa was confirmed by sequencing and increased H3K9me3.Whole-mount in situ hybridization showed that kdm4aa is widely expressed during the embryonic development of zebrafish.Compared with WT zebrafish,kdm4aa^(-/-)zebrafish showed no significant difference in gamete formation and fertilization,but the survival rate of kdm4aa^(-/-)embryos dramatically reduced to 21%at 26 hpf.Further observation showed that about 80%of survived kdm4aa^(-/-)zebrafish experienced disruption in stripe formation,and 10%of survived kdm4aa^(-/-)zebrafish underwent vertebral malformation.Alizarin red S staining demonstrated the abnormal spinal development in kdm4aa^(-/-)zebrafish.These results indicated that kdm4aa is required for normal embryonic development of zebrafish,loss of kdm4aa function leads to decreased survival during the early stages of zebrafish development and morphological variation in adult zebrafish.
