Paired box 3(Pax3)is a critical upstream regulator of the onset of myogenesis.We have previously identified two spliced isoforms ofpax3a(pax3a-1 andpax3a-2)and three spliced isoforms ofpax3b(pax3b-1,pax3b-2,and pax3b-3)in olive flounder,but their roles in myogenesis are unknown.In this study,we investigated their cellular localization,transcriptional activity on myod gene regulation,and roles in myogenesis.Different Pax3a and Pax3b isoforms revealed various subcellular localizations,which were re-lated to their corresponding protein structures.Pax3a-1,Pax3a-2,and Pax3b-1 promoted the transcriptional activity of myod to dif-ferent degrees,whereas Pax3b-2 and Pax3b-3 had a slight inhibitory or no effect.The pairwise interaction analysis demonstrated the synergistic effect of Pax3b-1 and Pax3b-3 on myod transcriptional activity.The overexpression of different pax3a and pax3b iso-forms differentially altered the spatial expression patterns of myod and differentially regulated the expression levels of their target genes(myod,myf5,and c-met)in zebrafish embryonic myogenesis.In addition,the different flounder myod promoter-driven pax3a/3b isoform expression vectors were successfully introduced into the skeletal muscles of juvenile flounder by electroporation.How-ever,none of them could change the mRNA expression levels of mstn,myf5,myod,myogenin,pax7a,and pax7b in the electroporated muscles.These results suggest that different Pax3a and Pax3b isoforms may precisely and collaboratively regulate embryonic myo-genesis,but their roles in juvenile myogenesis are uncertain.

JIAO Shuang;TAN Xungang;YOU Feng;PANG Qiuxiang

Anti-Aging&Regenerative Medicine Research Institution,School of Life Sciences and Medicine,Shandong University of Technology,Zibo 255000,China;Laboratory of Developmental and Evolutionary Biology,School of Life Sciences and Medicine,Shandong University of Technology,Zibo 255000,China;CAS and Shandong Province Key Laboratory of Experimental Marine Biology,Center for Ocean Mega-Science,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China;Laboratory for Marine Biology and Biotechnology,Pilot National Laboratory for Marine Science and Technology(Qingdao),Qingdao 266237,China

中国海洋大学学报（自然科学英文版）

[1]JIAO Shuang;TAN Xungang;YOU Feng;PANG Qiuxiang-.Multiple Isoforms of Olive Flounder(Paralichthys olivaceus)Pax3a and Pax3b Display Differential Regulations on Myogenic Differentiation)[J].中国海洋大学学报（自然科学英文版）,2022(05):1295-1306

Isoforms,Flounder,Pax3a,Pax3b,Myogenic,ofpax3a,pax3a,andpax3a,ofpax3b,pax3b,flounder,myod,myf5,pax7a,pax7b,electroporated

Multiple,Olive,Paralichthys,olivaceus,Display,Differential,Regulations,Differentiation,Paired,box,critical,upstream,regulator,onset,myogenesis,We,have,previously,identified,two,spliced,isoforms,three,olive,but,their,roles,are,unknown,In,this,study,investigated,transcriptional,activity,regulation,revealed,various,subcellular,localizations,which,were,corresponding,protein,structures,promoted,degrees,whereas,had,slight,inhibitory,effect,pairwise,interaction,analysis,demonstrated,synergistic,overexpression,differentially,altered,spatial,patterns,regulated,levels,target,met,zebrafish,embryonic,addition,promoter,driven,vectors,successfully,introduced,into,skeletal,muscles,juvenile,by,electroporation,How,ever,none,them,could,change,mstn,myogenin,These,results,suggest,that,may,precisely,collaboratively,uncertain

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