Knockdown of ttc26 disrupts ciliogenesis of the photoreceptor cells and the pronephros in zebrafish.
In our effort to understand genetic disorders of the photoreceptor cells of the retina, we have focused on intraflagellar transport in photoreceptor sensory cilia. From previous mouse proteomic data we identified a cilia protein Ttc26, orthologue of dyf-13 in Caenorhabditis elegans, as a target. We localized Ttc26 to the transition zone of photoreceptor and to the transition zone of cilia in cultured murine inner medullary collecting duct 3 (mIMCD3) renal cells. Knockdown of Ttc26 in mIMCD3 cells produced shortened and defective primary cilia, as revealed by immunofluorescence and scanning electron microscopy. To study Ttc26 function in sensory cilia in vivo, we utilized a zebrafish vertebrate model system. Morpholino knockdown of ttc26 in zebrafish embryos caused ciliary defects in the pronephric kidney at 27 h postfertilization and distension/dilation of pronephros at 5 d postfertilization (dpf). In the eyes, the outer segments of photoreceptor cells appeared shortened or absent, whereas cellular lamination appeared normal in retinas at 5 dpf. This suggests that loss of ttc26 function prevents normal ciliogenesis and differentiation in the photoreceptor cells, and that ttc26 is required for normal development and differentiation in retina and pronephros. Our studies support the importance of Ttc26 function in ciliogenesis and suggest that screening for TTC26 mutations in human ciliopathies is justified.
Ocular Genomics Institute & Berman-Gund Laboratory, Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA 02114, USA.
Qi
Q
Zhang
Qin
Q
Liu
Chrissy
C
Austin
Iain
I
Drummond
Eric A
EA
Pierce
D000818:Animals; D002460:Cell Line; D002923:Cilia / Q000502:physiology*; D055785:Gene Knockdown Techniques; D007814:Larva / Q000166:cytology; D008297:Male; D051379:Mice; D060172:Morpholinos / Q000235:genetics; D010641:Phenotype; D010786:Photoreceptor Cells / Q000166:cytology* / Q000378:metabolism; D060910:Pronephros / Q000166:cytology* / Q000378:metabolism; D021381:Protein Transport; D034622:RNA Interference; D051381:Rats; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
D060172:Morpholinos; C577010:Ttc26 protein, zebrafish; D029961:Zebrafish Proteins