Defect of centriole or cilia genes were highlighted as associated with inherited retinal diseases and ciliopathy, which has a broad spectrum of phenotype including vision impairment. The mechanism may be interrupt of cilia formation. poc1b is an abundant protein in centriole proteomics, and it was found a novel gene for inherited retinal diseases.
Purpose Our aim is to study the role of poc1b in the photoreceptor sensory cilia formation and importance in vision function.
Methods Knock down of poc1b in mIMCD3 cell was used to study its role in general cilia formation. Gene blast, RT-PCR and western blot were used to study the conserved expression of poc1b. V5-poc1b and Flag-poc1b expression plasmids were constructed; and then in vitro (cell culture) and in vivo (rat sub-retinal injection) poc1b plasmid transfection were performed to detect location of poc1b in cilia and photoreceptor sensory cilia. For function study of poc1b, zebrafish gene knockdown, vision test, histology were processed.
Results poc1b was critical for cilia formation in mIMCD3 cell; poc1b was conserved expressed and was located in the basal body of mIMCD3 cell and mouse photoreceptor cilia; knockdown of poc1b leads to a spectrum phenotypes associated with cilia defect including vision impairment. Histology results show abnormal photoreceptor sensory cilia in defected larvae.
Conclusion poc1b is required for photoreceptor sensory cilia formation and normal vision in zebrafish.