title: The abcc6a gene expression is required for normal zebrafish development.
abstract: Pseudoxanthoma elasticum (PXE) is caused by mutations in the ABCC6 gene, which encodes a putative efflux transporter, ABCC6. The zebrafish (Danio rerio) has two ABCC6-related sequences. To study the function of abcc6 during zebrafish development, the mRNA expression levels were measured using RT-PCR and in situ hybridization. The abcc6a showed a relatively high level of expression at 5 days post-fertilization (d.p.f.) and the expression was specific to the Kupffer's vesicles. The abcc6b expression was evident at 6 hours post-fertilization (h.p.f.) and remained high up to 8 d.p.f., corresponding to embryonic kidney proximal tubules. Morpholinos were designed to both genes to prevent pre-mRNA splicing and block translation. Injection of the abcc6a morpholinos into 1-4 cell zebrafish embryos decreased gene expression by 54-81%, and induced a phenotype, pericardial edema and curled tail associated with death at around 8 d.p.f. Microinjecting zebrafish embryos with full-length mouse Abcc6 mRNA together with the morpholino completely rescued this phenotype. No phenotypic changes were observed when the abcc6b gene morpholino was injected into embryos with knock-down efficiency of 100%. These results suggest that abcc6a is an essential gene for normal zebrafish development and provide insight into the function of ABCC6, the gene mutated in PXE.
authors.affiliation: Department of Dermatology and Cutaneous Biology, Jefferson Medical College and Jefferson Institute of Molecular Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors.forename: Qiaoli
authors.initials: Q
authors.lastname: Li
authors.forename: Sara
authors.initials: S
authors.lastname: Sadowski
authors.forename: Michael
authors.initials: M
authors.lastname: Frank
authors.forename: Chunli
authors.initials: C
authors.lastname: Chai
authors.forename: Andras
authors.initials: A
authors.lastname: Váradi
authors.forename: Shiu-Ying
authors.initials: SY
authors.lastname: Ho
authors.forename: Hong
authors.initials: H
authors.lastname: Lou
authors.forename: Michael
authors.initials: M
authors.lastname: Dean
authors.forename: Christine
authors.initials: C
authors.lastname: Thisse
authors.forename: Bernard
authors.initials: B
authors.lastname: Thisse
authors.forename: Jouni
authors.initials: J
authors.lastname: Uitto
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D055785:Gene Knockdown Techniques; D007687:Kidney Tubules, Proximal / Q000196:embryology / Q000502:physiology; D051379:Mice; D023421:Models, Animal; D008969:Molecular Sequence Data; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009841:Oligonucleotides / Q000235:genetics / Q000494:pharmacology; D010641:Phenotype; D010802:Phylogeny; D011561:Pseudoxanthoma Elasticum / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009841:Oligonucleotides; D029961:Zebrafish Proteins; C553407:ABCC6A protein, zebrafish; C504743:Abcc6 protein, mouse; C553408:abcc6b.1 protein, zebrafish