title: A pair of Sox: distinct and overlapping functions of zebrafish sox9 co-orthologs in craniofacial and pectoral fin development.
abstract: Understanding how developmental systems evolve after genome amplification is important for discerning the origins of vertebrate novelties, including neural crest, placodes, cartilage and bone. Sox9 is important for the development of these features, and zebrafish has two co-orthologs of tetrapod SOX9 stemming from an ancient genome duplication event in the lineage of ray-fin fish. We have used a genotype-driven screen to isolate a mutation deleting sox9b function, and investigated its phenotype and genetic interactions with a sox9a null mutation. Analysis of mutant phenotypes strongly supports the interpretation that ancestral gene functions partitioned spatially and temporally between Sox9 co-orthologs. Distinct subsets of the craniofacial skeleton, otic placode and pectoral appendage express each gene, and are defective in each single mutant. The double mutant phenotype is additive or synergistic. Ears are somewhat reduced in each single mutant but are mostly absent in the double mutant. Loss-of-function animals from mutations and morpholino injections, and gain-of-function animals injected with sox9a and sox9b mRNAs showed that sox9 helps regulate other early crest genes, including foxd3, sox10, snai1b and crestin, as well as the cartilage gene col2a1 and the bone gene runx2a; however, tfap2a was nearly unchanged in mutants. Chondrocytes failed to stack in sox9a mutants, failed to attain proper numbers in sox9b mutants and failed in both morphogenetic processes in double mutants. Pleiotropy can cause mutations in single copy tetrapod genes, such as Sox9, to block development early and obscure later gene functions. By contrast, subfunction partitioning between zebrafish co-orthologs of tetrapod genes, such as sox9a and sox9b, can relax pleiotropy and reveal both early and late developmental gene functions.
authors.affiliation: Institute of Neuroscience, University of Oregon, Eugene, OR 97403, USA.
authors.forename: Yi-Lin
authors.initials: YL
authors.lastname: Yan
authors.forename: John
authors.initials: J
authors.lastname: Willoughby
authors.forename: Dong
authors.initials: D
authors.lastname: Liu
authors.forename: Justin Gage
authors.initials: JG
authors.lastname: Crump
authors.forename: Catherine
authors.initials: C
authors.lastname: Wilson
authors.forename: Craig T
authors.initials: CT
authors.lastname: Miller
authors.forename: Amy
authors.initials: A
authors.lastname: Singer
authors.forename: Charles
authors.initials: C
authors.lastname: Kimmel
authors.forename: Monte
authors.initials: M
authors.lastname: Westerfield
authors.forename: John H
authors.initials: JH
authors.lastname: Postlethwait
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019521:Body Patterning; D001842:Bone and Bones / Q000196:embryology; D002356:Cartilage / Q000196:embryology; D016923:Cell Death; D019902:Chondrocytes / Q000378:metabolism; D004423:Ear / Q000196:embryology; D047108:Embryonic Development; D019143:Evolution, Molecular; D005121:Extremities; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D006609:High Mobility Group Proteins / Q000096:biosynthesis* / Q000502:physiology*; D017403:In Situ Hybridization; D020287:In Situ Nick-End Labeling; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D055758:SOX9 Transcription Factor; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis* / Q000502:physiology*; D015027:Zebrafish
chemical_list: D006609:High Mobility Group Proteins; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D014157:Transcription Factors