Zebrafish embryos possess two major types of myofibers,the slow and fast fibers,with distinct patterns of cell fusion.The fast muscle cells can fuse,while the slow muscle cells cannot.Here,we show that myomaker is exp...Zebrafish embryos possess two major types of myofibers,the slow and fast fibers,with distinct patterns of cell fusion.The fast muscle cells can fuse,while the slow muscle cells cannot.Here,we show that myomaker is expressed in both slow and fast muscle precursors,whereas myomixer is exclusive to fast muscle cells.The loss of Prdm1a,a regulator of slow muscle differentiation,results in strong myomaker and myomixer expression and slow muscle cell fusion.This abnormal fusion is further confirmed by the direct ectopic expression of myomaker or myomixer in slow muscle cells of transgenic models.Using the transgenic models,we show that the heterologous fusion between slow and fast muscle cells can alter slow muscle cell migration and gene expression.Furthermore,the overexpression of myomaker and myomixer also disrupts membrane integrity,resulting in muscle cell death.Collectively,this study identifies that the fiber-type-specific expression of fusogenic proteins is critical for preventing inappropriate fusion between slow and fast fibers in fish embryos,highlighting the need for precise regulation of fusogenic gene expression to maintain muscle fiber integrity and specificity.展开更多
The whole-genome sequence of the olive flounder(Paralichthys olivaceus)provides a basis for gene functional analyses,which is important for the aquaculture industry.Understanding gene function will help us to select b...The whole-genome sequence of the olive flounder(Paralichthys olivaceus)provides a basis for gene functional analyses,which is important for the aquaculture industry.Understanding gene function will help us to select better economic traits such as fast growth and better culture conditions,which further will increase the aquaculture output.Gene knockout is an important reverse genetics approach for in vivo studies of gene function.In this study,the CRISPR/Cas9 genome editing method with a microinjection system using a simple braked needle was employed in olive flounder.After injection in embryos,green fluorescent protein expression was detected in 40%of larvae.The proportion of normal-hatched larvae was approximately 50%.Different mutations,including short indels and fragment deletions,were found in our test genes gsdf and myomaker.Additionally,we detected more than one mutation in a single larva.In summary,our microinjection technique and CRISPR/Cas9 can be applied to study gene functions in olive flounder.展开更多
基金supported by funding from the U.S.National Institute of Health(NIH)National Institute of Arthritis and Musculoskeletal and Skin Diseases(R01AR072703 to S.D.)。
文摘Zebrafish embryos possess two major types of myofibers,the slow and fast fibers,with distinct patterns of cell fusion.The fast muscle cells can fuse,while the slow muscle cells cannot.Here,we show that myomaker is expressed in both slow and fast muscle precursors,whereas myomixer is exclusive to fast muscle cells.The loss of Prdm1a,a regulator of slow muscle differentiation,results in strong myomaker and myomixer expression and slow muscle cell fusion.This abnormal fusion is further confirmed by the direct ectopic expression of myomaker or myomixer in slow muscle cells of transgenic models.Using the transgenic models,we show that the heterologous fusion between slow and fast muscle cells can alter slow muscle cell migration and gene expression.Furthermore,the overexpression of myomaker and myomixer also disrupts membrane integrity,resulting in muscle cell death.Collectively,this study identifies that the fiber-type-specific expression of fusogenic proteins is critical for preventing inappropriate fusion between slow and fast fibers in fish embryos,highlighting the need for precise regulation of fusogenic gene expression to maintain muscle fiber integrity and specificity.
基金the National Natural Sci-ence Foundation of China(Nos.31672636,31772834,and 31972774)the National Key R&D Program of China(Nos.2018YFD0901202 and 2018YFD0900202)the Key Research and Development Program of Shandong Pro-vince,China(No.2019GHY1120070)。
文摘The whole-genome sequence of the olive flounder(Paralichthys olivaceus)provides a basis for gene functional analyses,which is important for the aquaculture industry.Understanding gene function will help us to select better economic traits such as fast growth and better culture conditions,which further will increase the aquaculture output.Gene knockout is an important reverse genetics approach for in vivo studies of gene function.In this study,the CRISPR/Cas9 genome editing method with a microinjection system using a simple braked needle was employed in olive flounder.After injection in embryos,green fluorescent protein expression was detected in 40%of larvae.The proportion of normal-hatched larvae was approximately 50%.Different mutations,including short indels and fragment deletions,were found in our test genes gsdf and myomaker.Additionally,we detected more than one mutation in a single larva.In summary,our microinjection technique and CRISPR/Cas9 can be applied to study gene functions in olive flounder.
