Zfp503/Nlz2 Is Required for RPE Differentiation and Optic Fissure Closure.
Zfp503/Nlz2 Is Required for RPE Differentiation and Optic Fissure Closure.
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DOI：
10.1167/iovs.63.12.5
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发表时间：
2022-11-01
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
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葡萄膜缺损是一种先天性眼部畸形，在人类早期发育过程中，由于视裂未能闭合而导致。尽管在确定其调节对于执行该闭合很重要的基因方面取得了重大进展，但使用已知基因组在少数情况下检测到突变，这意味着额外的遗传复杂性。我们先前已经证明了在斑马鱼中敲低znf 503（小鼠Zfp 503的直系同源物）会导致缺损。在这里，我们表征Zfp 503敲除（KO）小鼠，并评估突变体与野生型（WT）视网膜色素上皮（RPE）/脉络膜的转录组学分析。 通过使用同源重组的基因靶向产生Zfp 503 KO小鼠。对胚胎进行大体和组织学表征。用免疫染色/原位杂交检测发育相关蛋白/基因的模式和水平。将E11.5 KO RPE/脉络膜的转录组谱与WT的转录组谱进行比较。 zfp 503在发育中的小鼠眼睛中动态表达，其表达缺失导致葡萄膜缺损。KO胚胎表现出参与眼睛发育的几种关键转录因子的mRNA水平和表达模式的改变，包括Otx 2、Mitf、Pax 6、Pax 2、Vax 1和Vax 2，导致无法维持假定的RPE，如黑色素沉着减少及其分化为神经视网膜样谱系所证明的。WT和KO E11.5胚胎的RNA测序数据的比较表明黑色素相关基因的表达减少，并且与已知在视裂处动态调节的基因显著重叠。这些结果证明了Zfp 503在维持RPE命运和视裂闭合中的关键作用。
Uveal coloboma is a congenital eye malformation caused by failure of the optic fissure to close in early human development. Despite significant progress in identifying genes whose regulation is important for executing this closure, mutations are detected in a minority of cases using known gene panels, implying additional genetic complexity. We have previously shown knockdown of znf503 (the ortholog of mouse Zfp503) in zebrafish causes coloboma. Here we characterize Zfp503 knockout (KO) mice and evaluate transcriptomic profiling of mutant versus wild-type (WT) retinal pigment epithelium (RPE)/choroid. Zfp503 KO mice were generated by gene targeting using homologous recombination. Embryos were characterized grossly and histologically. Patterns and level of developmentally relevant proteins/genes were examined with immunostaining/in situ hybridization. The transcriptomic profile of E11.5 KO RPE/choroid was compared to that of WT. Zfp503 is dynamically expressed in developing mouse eyes, and loss of its expression results in uveal coloboma. KO embryos exhibit altered mRNA levels and expression patterns of several key transcription factors involved in eye development, including Otx2, Mitf, Pax6, Pax2, Vax1, and Vax2, resulting in a failure to maintain the presumptive RPE, as evidenced by reduced melanin pigmentation and its differentiation into a neural retina–like lineage. Comparison of RNA sequencing data from WT and KO E11.5 embryos demonstrated reduced expression of melanin-related genes and significant overlap with genes known to be dynamically regulated at the optic fissure. These results demonstrate a critical role of Zfp503 in maintaining RPE fate and optic fissure closure.
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影响因子：
3.7
作者：
Ou J;Bharti K;Nodari A;Bertuzzi S;Arnheiter H
通讯作者：
Arnheiter H
影响因子：
2.7
作者：
Dakubo, Gabriel D.;Mazerolle, Chantal;Wallace, Valerie A.
通讯作者：
Wallace, Valerie A.
影响因子：
5.6
作者：
Eckert, Priska;Knickmeyer, Max D.;Heermann, Stephan
通讯作者：
Heermann, Stephan
影响因子：
2.5
作者：
Hoyle, J;Tang, YP;Sive, H
通讯作者：
Sive, H
影响因子：
5.5
作者：
Chan BHC;Moosajee M;Rainger J
通讯作者：
Rainger J