title: A possible zebrafish model of polycystic kidney disease: knockdown of wnt5a causes cysts in zebrafish kidneys.
abstract: Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The molecular mechanisms leading to cyst formation in PKD remain somewhat unclear, but many genes are thought to be involved. Wnt5a is a non-canonical glycoprotein that regulates a wide range of developmental processes. Wnt5a works through the planar cell polarity (PCP) pathway that regulates oriented cell division during renal tubular cell elongation. Defects of the PCP pathway have been found to cause kidney cyst formation. Our paper describes a method for developing a zebrafish cystic kidney disease model by knockdown of the wnt5a gene with wnt5a antisense morpholino (MO) oligonucleotides. Tg(wt1b:GFP) transgenic zebrafish were used to visualize kidney structure and kidney cysts following wnt5a knockdown. Two distinct antisense MOs (AUG - and splice-site) were used and both resulted in curly tail down phenotype and cyst formation after wnt5a knockdown. Injection of mouse Wnt5a mRNA, resistant to the MOs due to a difference in primary base pair structure, rescued the abnormal phenotype, demonstrating that the phenotype was not due to "off-target" effects of the morpholino. This work supports the validity of using a zebrafish model to study wnt5a function in the kidney.
authors.affiliation: Department of Medicine, Eastern Virginia Medical School; HuangLE@evms.edu.
authors.forename: Liwei
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Medicine, Eastern Virginia Medical School.
authors.forename: An
authors.initials: A
authors.lastname: Xiao
authors.affiliation: Department of Medicine, Eastern Virginia Medical School.
authors.forename: Andrea
authors.initials: A
authors.lastname: Wecker
authors.affiliation: Department of Medicine, Eastern Virginia Medical School.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: McBride
authors.affiliation: Department of Medicine, Medical University of South Carolina.
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
authors.affiliation: Department of Pediatrics, University of Michigan.
authors.forename: Weibin
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Department of Medicine, Medical University of South Carolina.
authors.forename: Joshua H
authors.initials: JH
authors.lastname: Lipschutz
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004195:Disease Models, Animal; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000235:genetics; D007690:Polycystic Kidney Diseases / Q000235:genetics* / Q000378:metabolism; D051153:Wnt Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D000071818:Wnt-5a Protein; D015027:Zebrafish
chemical_list: D016376:Oligonucleotides, Antisense; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C438615:Wnt5a protein, mouse; D049452:Green Fluorescent Proteins