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Mingzhe Cao, Shuyi Chen; Afap1l2 controls the optic fissure closure in zebrafish. Invest. Ophthalmol. Vis. Sci. 2016;57(12):No Pagination Specified.
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© ARVO (1962-2015); The Authors (2016-present)
Ocular coloboma is a major cause for childhood blindness, which is caused by failure of the optic fissure (OF) closure during the eye morphogenesis. Currectly, the regulatory mechanisms underlying the OF closure remain largely undefined, hindering the diagnosis and treatment of the disease. In this study, we used zebrafish as the model system to study the function of afap1l2, an OF specifically expressed gene, in OF closure, with the hope of further elucidating the etiology of coloboma.
Whole-mount zebrafish embryo in situ hybridization was used to examine the dynamic expression patterns of afap1l2 during zebrafish OF development. Morpholinos and mRNA injection was used to study the function of afap1l2 in zebrafish OF closure. Whole-mount or cryo-section in situ hybridization and immunostaining was used to analyze gene expression changes in afap1l2 knockdown morphant fish. Images are analyzed under LEICA stereo- and ZEISS confocal microscopes.
24-72hpf zebrafish embryos whole-mount in situ hybridization results showed that afap1l2 is specifically expressed in the neural retina surrounding the closing OF at 48hpf. In addition, afap1l2 is also expressed along the optic stalk and the midline of the brain. Morpholino knockdown and mRNA rescue experiments showed that afap1l2 knockdown caused 55.7% to 58% zebrafish developed coloboma, which can be rescued by co-injecting afap1l2 mRNA, demonstrating that afap1l2 is required for the closure of the OF in zebrafish. Gene expression analysis showed that Pax2.1 expression was upregulated at the OF area, and shh was also upregulated at the forebrain midline.
afap1l2 is required for the OF closure in zebrafish. afap1l2 probably regulates the OF closure through controlling the forebrain midline shh signaling.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
afap1l2 is specifically expressed in the neural retina surrounding the closing OF at 48hpf.
afap1l2 knockdown morphant fish developed coloboma phenotype.
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