Knockdown of zebrafish crim1 results in a bent tail phenotype with defects in somite and vascular development.
The Crim1 gene encodes a transmembrane protein containing six cysteine-rich repeats similar to those found in the BMP antagonist, chordin (chd). To investigate its physiological role, zebrafish crim1 was cloned and shown to be both maternally and zygotically expressed during zebrafish development in sites including the vasculature, intermediate cell mass, notochord, and otic vesicle. Bent or hooked tails with U-shaped somites were observed in 85% of morphants from 12 hpf. This was accompanied by a loss of muscle pioneer cells. While morpholino knockdown of crim1 showed some evidence of ventralisation, including expansion of the intermediate cell mass (ICM), reduction in head size bent tails and disruption to the somites and notochord, this did not mimic the classically ventralised phenotype, as assessed by the pattern of expression of the dorsal markers chordin, otx2 and the ventral markers eve1, pax2.1, tal1 and gata1 between 75% epiboly and six-somites. From 24 hpf, morphants displayed an expansion of the ventral mesoderm-derived ICM, as evidenced by expansion of tal1, lmo2 and crim1 itself. Analysis of the crim1 morphant phenotype in Tg(fli:EGFP) fish showed a clear reduction in the endothelial cells forming the intersegmental vessels and a loss of the dorsal longitudinal anastomotic vessel (DLAV). Hence, the primary role of zebrafish crim1 is likely to be the regulation of somitic and vascular development.
Kinna
Genevieve
G
Institute for Molecular Bioscience, The University of Queensland, Brisbane, Qld 4072, Australia.
Kolle
Gabriel
G
Carter
Adrian
A
Key
Brian
B
Lieschke
Graham J
GJ
Perkins
Andrew
A
Little
Melissa H
MH
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001808:Blood Vessels / Q000196:embryology*; D019521:Body Patterning / Q000235:genetics; D052004:Bone Morphogenetic Protein Receptors / Q000235:genetics / Q000502:physiology*; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000235:genetics; D016376:Oligonucleotides, Antisense; D010641:Phenotype; D017385:Sequence Homology; D019170:Somites / Q000166:cytology*; D013623:Tail / Q000196:embryology*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000502:physiology; D029961:Zebrafish Proteins / Q000235:genetics / Q000502:physiology*
D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D029961:Zebrafish Proteins; C512401:crim1 protein, zebrafish; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D052004:Bone Morphogenetic Protein Receptors