title: Maternally supplied Smad5 is required for ventral specification in zebrafish embryos prior to zygotic Bmp signaling.
abstract: We have previously shown that the maternal effect dorsalization of zebrafish embryos from sbn(dtc24) heterozygous mothers is caused by a dominant negative mutation in Smad5, a transducer of ventralizing signaling by the bone morphogenetic proteins Bmp2b and Bmp7. Since sbn(dtc24) mutant Smad5 protein not only blocks wild-type Smad5, but also other family members like Smad1, it remained open to what extent Smad5 itself is required for dorsoventral patterning. Here, we report the identification of novelsmad5 alleles: three new isolates coming from a dominant enhancer screen, and four former isolates initially assigned to the cpt and pgy complementation groups. Overexpression analyses demonstrate that three of the new alleles, m169, fr5, and tc227, are true nulls (amorphs), whereas the initial dtc24 allele is both antimorphic and hypomorphic. We rescued m169 mutant embryos by smad5 mRNA injection. Although adult mutants are smaller than their siblings, the eggs laid by m169(-/-) females are larger than normal eggs. Embryos lacking maternal Smad5 function (Mm169(-/-) embryos) are even more strongly dorsalized thanbmp2b or bmp7 null mutants. They do not respond to injected bmp2b mRNA, indicating that Smad5 is absolutely essential for ventral development and Bmp2/7 signaling. Most importantly, Mm169(-/-) embryos display reducedbmp7 mRNA levels during blastula stages, when bmp2b and bmp7 mutants are still normal. This indicates that maternally supplied Smad5 is already required to mediate ventral specification prior to zygotic Bmp2/7 signaling to establish the initial dorsoventral asymmetry.
authors.affiliation: Max-Planck Institute for Immunobiology, Stuebeweg 51, D-79108 Freiburg, Germany.
authors.forename: Carina
authors.initials: C
authors.lastname: Kramer
authors.forename: Thomas
authors.initials: T
authors.lastname: Mayr
authors.forename: Matthias
authors.initials: M
authors.lastname: Nowak
authors.forename: Jennifer
authors.initials: J
authors.lastname: Schumacher
authors.forename: Greg
authors.initials: G
authors.lastname: Runke
authors.forename: Hermann
authors.initials: H
authors.lastname: Bauer
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Wagner
authors.forename: Bettina
authors.initials: B
authors.lastname: Schmid
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Imai
authors.forename: William S
authors.initials: WS
authors.lastname: Talbot
authors.forename: Mary C
authors.initials: MC
authors.lastname: Mullins
authors.forename: Matthias
authors.initials: M
authors.lastname: Hammerschmidt
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019521:Body Patterning / Q000235:genetics / Q000502:physiology; D055396:Bone Morphogenetic Protein 2; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000502:physiology*; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology*; D004742:Enhancer Elements, Genetic; D005260:Female; D005816:Genetic Complementation Test; D006720:Homozygote; D008969:Molecular Sequence Data; D009154:Mutation; D009866:Oogenesis / Q000235:genetics / Q000502:physiology; D010641:Phenotype; D010750:Phosphoproteins / Q000235:genetics* / Q000502:physiology*; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051902:Smad5 Protein; D015534:Trans-Activators / Q000235:genetics* / Q000502:physiology*; D016212:Transforming Growth Factor beta; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics* / Q000502:physiology; D029961:Zebrafish Proteins; D015053:Zygote / Q000254:growth & development
chemical_list: D055396:Bone Morphogenetic Protein 2; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D004268:DNA-Binding Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D051902:Smad5 Protein; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D029961:Zebrafish Proteins; C405936:bmp2b protein, zebrafish; C525826:bmp7a protein, zebrafish; C497801:smad5 protein, zebrafish; D004247:DNA