State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Guangdong Province Key Laboratory of Brain Function and Disease, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
Invest Ophthalmol Vis Sci. 2018 Mar 1;59(3):1167-1177. doi: 10.1167/iovs.17-22827.
Coloboma is a sight-threatening congenital eye disease caused by a failure in optic fissure (OF) closure. The aim of this study was to investigate the role of Adamts16, a metalloproteinase, in OF closure.
RNA in situ hybridization was used to examine the expression of Adamts16 in developing mouse and zebrafish eyes. Morpholino knockdowns were performed to study adamts16 function during zebrafish eye development. Additionally, immunofluorescent staining, RNA in situ hybridization, bromodeoxyuridine (BrdU) labeling, TUNEL assays, and high-throughput sequencing were used to examine altered cellular and molecular events in adamts16-morphant optic cups (OCs).
Adamts16 is expressed at the edges of the closing OF in both mice and zebrafish eyes. Zebrafish adamts16 knockdown resulted in coloboma formation. In adamts16-morphant eyes, the basement membrane failed to disassemble at the closing OF edges, OC cells exhibited decreased proliferation and increased apoptosis, and fibroblast growth factor 8 (fgf8) was ectopically upregulated in the OC.
adamts16 is required for proper OF closure in zebrafish eyes. adamts16 controls OF closure possibly through the combined functions of degrading the basement membrane at the closing OF edges, promoting cell proliferation and survival, and restricting fgf8 expression. Our study linked a metalloproteinase to OF closure, which may facilitate future etiologic studies on human coloboma cases.
视窝是一种由视裂（OF）闭合失败引起的威胁视力的先天性眼部疾病。本研究旨在探讨金属蛋白酶 Adamts16 在 OF 闭合中的作用。
采用 RNA 原位杂交技术检测 Adamts16 在发育中小鼠和斑马鱼眼中的表达。进行形态发生素敲低实验以研究 adamts16 在斑马鱼眼部发育过程中的功能。此外，还进行了免疫荧光染色、RNA 原位杂交、溴脱氧尿苷（BrdU）标记、TUNEL 检测和高通量测序，以研究 adamts16 突变体视杯（OC）中改变的细胞和分子事件。
Adamts16 在小鼠和斑马鱼眼睛的闭合 OF 边缘表达。斑马鱼 adamts16 敲低导致视窝形成。在 adamts16 突变体眼睛中，基底膜在闭合 OF 边缘未能解体，OC 细胞增殖减少，凋亡增加，OC 中纤维母细胞生长因子 8（fgf8）异位上调。
adamts16 是斑马鱼眼睛中 OF 正常闭合所必需的。adamts16 通过降解闭合 OF 边缘的基底膜、促进细胞增殖和存活以及限制 fgf8 表达的综合功能来控制 OF 闭合。我们的研究将一种金属蛋白酶与 OF 闭合联系起来，这可能有助于未来对人类视窝病例的病因研究。
