Pediatric Nephrology

, Volume 23, Issue 3, pp 377–387 | Cite as

Defects in ciliary localization of Nek8 is associated with cystogenesis

  • Melissa L. Trapp
  • Alevtina Galtseva
  • Danielle K. Manning
  • David R. Beier
  • Norman D. Rosenblum
  • Lynne M. Quarmby
Original Article


Mutations in the human NIMA (Never in Mitosis gene A)-related kinase 8 (Nek8) are associated with a rare form of the juvenile renal cystic disease, nephronophthisis type 9, and mutations in murine Nek8 cause renal cysts in jck mice. Cystogenesis involves dysfunctional ciliary signaling, and we have previously reported that Nek8 localizes to the primary cilium in mouse kidney epithelial cells. We now report that in developing mouse kidney, Nek8 is detected in the cilia of a subset of ureteric-bud-derived tubules at embryonic day (E)15.5. An increasing proportion of ureteric-bud-derived tubules express ciliary Nek8 until E18.5. Postnatal day 1 and 7 Nek8 is observed with equal frequency in both ureteric-bud and non-ureteric-bud-derived tubules. To investigate the cell biological consequences of kinase-deficient and jck mutant forms of Nek8, we transiently expressed green fluorescent protein (GFP)-tagged constructs in vitro. Mutations in the kinase and C-terminal domains of Nek8 adversely affected ciliary targeting but did not affect ciliogenesis or ciliary length. Consistent with these in vitro observations, kidneys from homozygous jck mice revealed reduced ciliary expression of Nek8 compared with kidneys from heterozygous (unaffected) mice. These data indicate that the ciliary localization of Nek8 in a subset of ureteric-bud-derived kidney tubules is essential for maintaining the integrity of those tubules in the mammalian kidney.


Polycystic kidney disease Nephronophthisis Cilia Kinases 



We thank Lin Chen of the Rosenblum lab for invaluable assistance. We also thank Michel Leroux and his lab members for the use of their tissue culture facilities. MLT was supported by graduate fellowships from the Michael Smith Foundation for Health Research and the Natural Sciences and Engineering Research Council of Canada. Work in the Quarmby lab was supported by operating grants from the Kidney Foundation of Canada and by the Canadian Institutes of Health Research (MOP-37861). Work in the Rosenblum lab was supported by grants from the Canadian Institutes of Health Research and the Canada Research Chairs Program.

Supplementary material

467_2007_692_MOESM1_ESM.doc (1.1 mb)
Fig. S1 Condensing mesenchyme does not express NIMA (never in mitosis gene a)-related kinase 8 (Nek8). Paraffin-embedded sections of embryonic day (E)18.5 CD1/129 mouse kidneys stained with anti-Nek8 antibody, anti-neural-cell adhesion molecule (NCAM) antibody, and 4′,6-diamidino-2-phenylindole (DAPI) (blue). a Tubules in the cortex labeled with anti-NCAM (red) do not express luminal Nek8 (green). b NCAM-positive tubules (green) stained with anti-acetylated tubulin (red), indicating cilia (DOC 1.12 MB)
467_2007_692_MOESM2_ESM.doc (145 kb)
Fig. S2 In vitro ciliary measurements. Confluent inner medullary collecting duct (IMCD)-3 cells transfected with no DNA (mock), GFP alone, green fluorescent protein (GFP)-wild-type NIMA (never in mitosis gene a)-related kinase 8 (Nek8), GFP-K33M Nek8, and GFP-jck Nek8 were stained with anti-acetylated tubulin, and ciliary length was measured. Also shown is untransfected (untransf.) cells. The average of two trials is shown. Error bars = standard error of mean (DOC 145 KB)
467_2007_692_MOESM3_ESM.doc (6 mb)
Fig. S3 The jck renal morphology. Paraffin-embedded sections of postnatal day 7 jck heterozygous (+/jck) and homozygous (jck/jck) mouse kidneys from litter mates were stained with hematoxylin and eosin (DOC 6.00 MB)


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Copyright information

© IPNA 2007

Authors and Affiliations

  • Melissa L. Trapp
    • 1
  • Alevtina Galtseva
    • 2
  • Danielle K. Manning
    • 3
  • David R. Beier
    • 3
  • Norman D. Rosenblum
    • 2
    • 4
  • Lynne M. Quarmby
    • 1
  1. 1.Department of Molecular Biology and BiochemistrySimon Fraser UniversityBurnabyCanada
  2. 2.Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, Department of PhysiologyUniversity of TorontoTorontoCanada
  3. 3.Genetics Division, Brigham and Women’s HospitalHarvard Medical SchoolBostonUSA
  4. 4.Department of PaediatricsUniversity of TorontoTorontoCanada

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