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

Autor: Val C. Sheffield, Qihong Zhang, Alexander G. Bassuk, Trudi A. Westfall, Diane C. Slusarski, Xue Mei
Rok vydání: 2014
Předmět:
Retinal neurogenesis
Morpholinos
Mice
0302 clinical medicine
Cell Movement
Cell polarity
In Situ Hybridization
Zebrafish
Mice
Knockout
0303 health sciences
Gene knockdown
Microscopy
Confocal
Reverse Transcriptase Polymerase Chain Reaction
Cilium
Cell Polarity
LIM Domain Proteins
Immunohistochemistry
Cell biology
Prickle2
Neural Tube
congenital
hereditary
and neonatal diseases and abnormalities
Neurogenesis
Kupffer׳s vesicle
Biology
Article
Retina
03 medical and health sciences
Bardet–Biedl syndrome
Intraflagellar transport
Ciliogenesis
medicine
Animals
Immunoprecipitation
Cilia
Bardet-Biedl Syndrome
Molecular Biology
Adaptor Proteins
Signal Transducing
DNA Primers
030304 developmental biology
Analysis of Variance
Membrane Proteins
Biological Transport
Cell Biology
Planar cell polarity
medicine.disease
Molecular biology
Cytoskeletal Proteins
Ciliopathy
Melanosome transport
Intracellular transport
030217 neurology & neurosurgery
Molecular Chaperones
Developmental Biology
Zdroj: Developmental Biology. 392:245-255
ISSN: 0012-1606
DOI: 10.1016/j.ydbio.2014.05.020
Popis: 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.
Databáze: OpenAIRE
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