Wang Zhongxiao, Liu Chi-Hsiu, Huang Shuo, Chen Jing
Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School.
Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School; The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School;
J Vis Exp. 2019 May 15(147). doi: 10.3791/59222.
In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed regression of hyaloid vessels can be seen in diseases such as persistent hyperplastic primary vitreous (PHPV), leading to an obstructed light path and impaired visual function. Understanding the mechanisms underlying the hyaloid vessel regression may lead to new molecular insights into the vascular regression process and potential new ways to manage diseases with persistent hyaloid vessels. Here we describe the procedures for imaging hyaloid in live mice with optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) and a detailed technical protocol of isolating and flat-mounting hyaloid ex vivo for quantitative analysis. Low-density lipoprotein receptor-related protein 5 (LRP5) knockout mice were used as an experimental model of persistent hyaloid vessels, to illustrate the techniques. Together, these techniques may facilitate a thorough assessment of hyaloid vessels as an experimental model of vascular regression and studies on the mechanism of persistent hyaloid vessels.
在眼睛中,胚胎玻璃样血管滋养发育中的晶状体和视网膜,并在视网膜血管发育时退化。玻璃样血管持续存在或退化失败可见于诸如持续性增生性原发性玻璃体(PHPV)等疾病中,导致光路受阻和视觉功能受损。了解玻璃样血管退化的潜在机制可能会为血管退化过程带来新的分子见解,并为治疗玻璃样血管持续存在的疾病提供潜在的新方法。在这里,我们描述了用光学相干断层扫描(OCT)和眼底荧光血管造影(FFA)对活体小鼠的玻璃样血管进行成像的方法,以及离体分离和平铺玻璃样血管以进行定量分析的详细技术方案。使用低密度脂蛋白受体相关蛋白5(LRP5)基因敲除小鼠作为玻璃样血管持续存在的实验模型来说明这些技术。总之,这些技术可能有助于全面评估玻璃样血管作为血管退化的实验模型以及对玻璃样血管持续存在机制的研究。