Nephrocystin-3 is required for ciliary function in zebrafish embryos.
Nephronophthisis (NPHP) is the most frequent genetic cause of end-stage renal failure in the first three decades of life. It is characterized primarily by renal cysts with extrarenal involvements of the eye and brain. Ten recessive genes responsible for NPHP have been identified by positional cloning. This discovery supported a unifying theory of renal cystic disease, which states that all proteins mutated in cystic kidney diseases of human, mice, or zebrafish are expressed in primary cilia of renal epithelial cells. Mutations in nephrocystin-3 (NPHP3) are the cause of human nephronophthisis type 3 and polycystic kidney disease (pcy) mouse mutants. To study the functional role of NPHP3 in normal embryonic development and in the pathogenesis of cystic kidney disease, we characterized the zebrafish ortholog nphp3 by morpholino oligo (MO)-mediated knockdown. When nphp3 function was suppressed by either of the two MOs blocking the translation of the protein or the splicing of mRNA, zebrafish embryos displayed hydrocephalus and pronephric cysts. Knockdown of nphp3 also led to situs inversus phenotypes due to defective cilia at Kupffer's vesicle. We showed that nphp3 genetically interacts with nphp2/inversin and human NPHP3 localizes to primary cilia in Madin-Darby canine kidney cells. Like nphp2/inversin, nphp3 knockdown affected morphogenic cell movement during gastrulation, suggesting nphp3 is essential to regulate convergent extension. Thus nphp3, cooperating with nphp2/inversin, plays an essential role related to ciliary function, and the knockdown provides an animal model that may be used for studies of the pathogenesis and therapy for this disease.
Zhou
Weibin
W
Department of Pediatrics, University of Michigan, Ann Arbor, Michigan, USA.
Dai
Julie
J
Attanasio
Massimo
M
Hildebrandt
Friedhelm
F
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002923:Cilia / Q000378:metabolism*; D004285:Dogs; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000378:metabolism; D054262:Gastrulation; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D005838:Genotype; D006801:Humans; D006849:Hydrocephalus / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000002:abnormalities / Q000378:metabolism*; D052177:Kidney Diseases, Cystic / Q000235:genetics / Q000378:metabolism; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009025:Morpholines / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D010641:Phenotype; D012857:Situs Inversus / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D015027:Zebrafish / Q000002:abnormalities / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
D009025:Morpholines; D009841:Oligonucleotides; D014404:Tubulin; D029961:Zebrafish Proteins; C550737:nephrocystin-3, zebrafish; C528697:nephrocystin-3, human; D016547:Kinesins
J Am Soc Nephrol. 2006 Oct;17(10):2706-18
16943304
Development. 2004 Aug;131(16):4085-93
15269167
Nat Genet. 2007 Nov;39(11):1350-60
17906624
Semin Cell Dev Biol. 2002 Jun;13(3):251-60
12137734
Development. 2005 Mar;132(6):1247-60
15716348
Gene Expr Patterns. 2009 Jun;9(5):282-8
19332147
Nat Genet. 2005 Mar;37(3):282-8
15723066
Development. 2005 Apr;132(8):1907-21
15790966
Pediatr Nephrol. 2008 Nov;23(11):2095-9
18604564
Nat Genet. 2005 Oct;37(10):1135-40
16170314
J Pathol. 2004 Nov;204(4):470-7
15495266
Nature. 2010 Apr 15;464(7291):1048-51
20393563
Am J Hum Genet. 2003 Mar;72(3):650-8
12567324
Am J Kidney Dis. 2009 May;53(5):790-5
19303681
Kidney Int. 2009 Apr;75(8):839-47
19177160
Nat Genet. 2003 Aug;34(4):413-20
12872123
Hum Mutat. 2008 Mar;29(3):418-26
18076122
Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15765-70
17884984
Nat Med. 2003 Oct;9(10):1323-6
14502283
Mol Biol Cell. 2002 Sep;13(9):3096-106
12221118
Stain Technol. 1974 Jan;49(1):9-14
4131626
Am J Hum Genet. 2008 Apr;82(4):959-70
18371931
J Am Soc Nephrol. 2009 Jan;20(1):23-35
19118152
Development. 1999 Jun;126(14):3253-62
10375514
Development. 2003 Jun;130(11):2303-16
12702646
Hum Mol Genet. 2008 Dec 1;17(23):3655-62
18723859
Nature. 2006 Jan 19;439(7074):326-30
16327777
Dev Biol. 2008 Feb 15;314(2):261-75
18178183
Nat Genet. 2003 Aug;34(4):455-9
12872122
Dev Cell. 2008 Jun;14(6):954-61
18539122
Hum Mol Genet. 2006 Mar 1;15(5):667-77
16399798
Development. 2007 Apr;134(8):1605-15
17360770
Nat Genet. 2006 Feb;38(2):155-7
16415886
Nat Genet. 2006 Jun;38(6):674-81
16682973
J Am Soc Nephrol. 2007 Jun;18(6):1855-71
17513324
Nat Genet. 2005 May;37(5):537-43
15852005