PUBLICATION

Functional characterization of Prickle2 and BBS7 identify overlapping phenotypes yet distinct mechanisms

Authors
Mei, X., Westfall, T.A., Zhang, Q., Sheffield, V.C., Bassuk, A.G., Slusarski, D.C.
ID
ZDB-PUB-140619-9
Date
2014
Source
Developmental Biology   392(2): 245-55 (Journal)
Registered Authors
Slusarski, Diane C.
Keywords
Bardet–Biedl syndrome, Cilia, Intracellular transport, Kupffer’s vesicle, Planar cell polarity, Prickle2, Retinal neurogenesis, Zebrafish
MeSH Terms
  • Biological Transport/physiology
  • Morpholinos/genetics
  • Immunohistochemistry
  • Analysis of Variance
  • Mice, Knockout
  • Bardet-Biedl Syndrome/genetics*
  • Membrane Proteins/genetics
  • Membrane Proteins/metabolism*
  • Retina/embryology
  • Cell Movement/physiology
  • Cilia/pathology
  • Immunoprecipitation
  • Reverse Transcriptase Polymerase Chain Reaction
  • Microscopy, Confocal
  • Molecular Chaperones/metabolism*
  • Cell Polarity/physiology*
  • Mice
  • Animals
  • Neural Tube/cytology
  • Neural Tube/embryology*
  • DNA Primers/genetics
  • LIM Domain Proteins/genetics
  • LIM Domain Proteins/metabolism*
  • Zebrafish
  • In Situ Hybridization
  • Neurogenesis/physiology*
(all 26)
PubMed
24938409 Full text @ Dev. Biol.
Abstract
Ciliopathies are genetic disorders that are caused by dysfunctional cilia and affect multiple organs. One type of ciliopathy, Bardet-Biedl syndrome, is a rare disorder characterized by obesity, retinitis pigmentosa, polydactyly, mental retardation and susceptibility to cardiovascular diseases. The Wnt/Planar cell polarity (PCP) has been associated with cilia function and ciliogenesis in directing the orientation of cilia and basal bodies. Yet the exact relationship between PCP and ciliopathy is not well understood. Here, we examine interactions between a core PCP component, Prickle2 (Pk2), and a central BBS gene, Bbs7, using gene knockdown in the zebrafish. pk2 and bbs7 knockdown both disrupt the formation of a ciliated organ, the Kupffer׳s vesicle (KV), but do not display a synergistic interaction. By measuring cell polarity in the neural tube, we find that bbs7 activity is not required for Pk asymmetric localization. Moreover, BBS protein complex formation is preserved in the Pk2-deficient (Pk2(-/-)) mouse. Previously we reported an intracellular melanosome transport delay as a cardinal feature of reduced bbs gene activity. We find that pk2 knockdown suppresses bbs7-related retrograde transport delay. Similarly, knockdown of ift22, an anterograde intraflagellar transport component, also suppresses the bbs7-related retrograde delay. Notably, we find that pk2 knockdown larvae show a delay in anterograde transport. These data suggest a novel role for Pk2 in directional intracellular transport and our analyses show that PCP and BBS function independently, yet result in overlapping phenotypes when knocked down in zebrafish.
Genes / Markers
Figures
Figure Gallery (5 images)
Show all Figures
Expression
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay Figure
prickle2bWTstandard conditionsProtruding-mouthISH
prickle2bWTstandard conditionsProtruding-mouthISH
1 - 2 of 2
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Phenotype
Fish Conditions Stage Phenotype Figure
WT + MO1-bbs7standard conditions5-9 somites to 10-13 somites
WT + MO1-bbs7standard conditions10-13 somites
WT + MO1-bbs7standard conditions10-13 somites
WT + MO1-bbs7chemical treatment: caffeineDay 5
WT + MO1-bbs7constant dark, chemical treatment: (R)-adrenalineDay 5
WT + MO1-bbs7constant dark, chemical treatment: (R)-adrenalineDay 5
WT + MO1-bbs7 + MO3-ift122constant dark, chemical treatment: (R)-adrenalineDay 5
WT + MO1-bbs7 + MO3-prickle2bstandard conditions5-9 somites to 10-13 somites
WT + MO1-bbs7 + MO3-prickle2bstandard conditions10-13 somites
WT + MO1-bbs7 + MO3-prickle2bstandard conditions10-13 somites
1 - 10 of 23
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
knu3TgTransgenic Insertion
    1 - 1 of 1
    Show
    Human Disease / Model
    Human Disease Fish Conditions Evidence
    Bardet-Biedl syndromeTAS
    1 - 1 of 1
    Show
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    bbs7MO1-bbs7MRPHLNO
    ift122MO3-ift122MRPHLNO
    prickle2bMO3-prickle2bMRPHLNO
    1 - 3 of 3
    Show
    Fish
    Fish
    knu3Tg + MO3-prickle2b
    WT
    WT + MO1-bbs7
    WT + MO1-bbs7 + MO3-ift122
    WT + MO1-bbs7 + MO3-prickle2b
    WT + MO3-ift122
    WT + MO3-prickle2b
    1 - 7 of 7
    Show
    Antibodies
    No data available
    Orthology
    No data available
    Engineered Foreign Genes
    Marker Marker Type Name
    EGFPEFGEGFP
    1 - 1 of 1
    Show
    Mapping
    No data available
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    Citation
    Mei, X., Westfall, T.A., Zhang, Q., Sheffield, V.C., Bassuk, A.G., Slusarski, D.C. (2014) Functional characterization of Prickle2 and BBS7 identify overlapping phenotypes yet distinct mechanisms. Developmental Biology. 392(2):245-55.