Single-cell transcriptomes and runx2b^(-/-)mutants reveal the genetic signatures of intermuscular bone formation in zebrafish  

作　　者：Chun-Hong Nie Shi-Ming Wan Yu-Long Chen Ann Huysseune Ya-Ming Wu Jia-Jia Zhou Alexandre Wagner Silva Hilsdorf Wei-Min Wang Paul Eckhard Witten Qiang Lin Ze-Xia Gao 

机构地区：[1]College of Fisheries,Key Lab of Freshwater Animal Breeding,Ministry of Agriculture and Rural Affairs/Key Lab of Agricultural Animal Genetics,Breeding and Reproduction of Ministry of Education/Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt,Ministry of Education,Huazhong Agricultural University,Wuhan 430070,China [2]Hubei Hongshan Laboratory,Wuhan 430070,China [3]CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology,South China Sea Institute of Oceanology,Innovation Academy of South China Sea Ecology and Environmental Engineering,Chinese Academy of Sciences,Guangzhou 510301,China [4]Department of Biology,Ghent University,Ghent B-9000,Belgium [5]Unit of Biotechnology,University of Mogi das Cruzes,Mogi das Cruzes,Săao Paulo 08780-911,Brazil [6]Guangdong Laboratory for Lingnan Modern Agriculture,Guangzhou 510000,China

出　　处：《National Science Review》2022年第11期92-102,共11页国家科学评论（英文版）

基　　金：supported by the earmarked fund for China Agriculture Research System (CARS-45-01);Hubei Hongshan Laboratory (2021hszd001);National Natural Science Foundation of China (31872559, 41825013 and U21A20263);National Key Research and Development Program of China(2018YFD0900102);Laboratory for Lingnan Modern Agriculture (NZ2021011);Special Project of Science and Technology Innovation of the Ministry of Education (2021ZL08);Wuhan Applied Foundational Frontier Project (2020020601012253);Key Research Program of Frontier Sciences of CAS (ZDBSLY-DQC004)

摘　　要：Intermuscular bones(IBs)are mineralized spicules,present in the myosepta of many,but not all,teleost species.IBs are often small and sharp,and they consequently limit how the fish can be processed;the IBs may cause injury or trauma if lodged in consumers’throats or mouths,and therefore affect the appeal of the fish to many consumers.The development of IBs in teleosts is still not fully understood and the molecular basis of IB development remains to be established.Here,the characteristics of IB tissue are evaluated based on single-cell transcriptomics in wild-type zebrafish.The analysis defined 18 distinct cell types.Differentiation trajectories showed that IBs are derived from tendons and that a core tendon-osteoblast cell lineage is related to IB formation.In particular,the functions of 10 candidate genes were evaluated via CRISPR-Cas9 mutants.Among those,runx2b^(-/-)mutants completely lost IBs,while swimming performance,growth and bone mineral density were not significantly different from runx2b^(+/+)zebrafish.Comparative single-cell RNA sequencing(sc RNA-seq)analysis in runx2b^(-/-)and runx2b^(+/+)zebrafish revealed the role of osteoblasts in IB formation.In addition,differentially expressed genes were enriched in the transforming growth factorβ/bone morphogenetic protein(TGF-β/BMP)pathway after runx2b deletion.This study provides evidence for the crucial role of runx2b regulation in IB formation.Genetic breeding can target runx2b regulation and generate strains of commercial fish species without IBs,which can improve the safe consumption and economic value of many farmed fish species.

关 键 词：TELEOSTS ZEBRAFISH intermuscular bones scRNA-seq tendon-osteoblast cell lineage runx2b genetic breeding 

分 类 号：S917.4[农业科学—水产科学]