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用于远程高通量表型分析的小鼠眼球摘除术。

Mouse eye enucleation for remote high-throughput phenotyping.

作者信息

Mahajan Vinit B, Skeie Jessica M, Assefnia Amir H, Mahajan Maryann, Tsang Stephen H

机构信息

Department of Ophthalmology and Visual Sciences, University of Iowa, USA.

出版信息

J Vis Exp. 2011 Nov 19(57):3184. doi: 10.3791/3184.

Abstract

The mouse eye is an important genetic model for the translational study of human ophthalmic disease. Blinding diseases in humans, such as macular degeneration, photoreceptor degeneration, cataract, glaucoma, retinoblastoma, and diabetic retinopathy have been recapitulated in transgenic mice.(1-5) Most transgenic and knockout mice have been generated by laboratories to study non-ophthalmic diseases, but genetic conservation between organ systems suggests that many of the same genes may also play a role in ocular development and disease. Hence, these mice represent an important resource for discovering new genotype-phenotype correlations in the eye. Because these mice are scattered across the globe, it is difficult to acquire, maintain, and phenotype them in an efficient, cost-effective manner. Thus, most high-throughput ophthalmic phenotyping screens are restricted to a few locations that require on-site, ophthalmic expertise to examine eyes in live mice. (6-9) An alternative approach developed by our laboratory is a method for remote tissue-acquisition that can be used in large or small-scale surveys of transgenic mouse eyes. Standardized procedures for video-based surgical skill transfer, tissue fixation, and shipping allow any lab to collect whole eyes from mutant animals and send them for molecular and morphological phenotyping. In this video article, we present techniques to enucleate and transfer both unfixed and perfusion fixed mouse eyes for remote phenotyping analyses.

摘要

小鼠眼睛是人类眼科疾病转化研究的重要遗传模型。人类的致盲性疾病,如黄斑变性、光感受器变性、白内障、青光眼、视网膜母细胞瘤和糖尿病性视网膜病变,已在转基因小鼠中得到重现。(1-5)大多数转基因和基因敲除小鼠是由实验室培育出来用于研究非眼科疾病的,但器官系统之间的遗传保守性表明,许多相同的基因也可能在眼部发育和疾病中发挥作用。因此,这些小鼠是发现眼睛新基因型-表型相关性的重要资源。由于这些小鼠分布在全球各地,难以以高效、经济的方式获取、饲养并对其进行表型分析。因此,大多数高通量眼科表型筛选仅限于少数几个需要现场眼科专业知识来检查活体小鼠眼睛的地点。(6-9)我们实验室开发的一种替代方法是一种远程组织采集方法,可用于对转基因小鼠眼睛进行大规模或小规模调查。基于视频的手术技能传授、组织固定和运输的标准化程序使任何实验室都能够从突变动物身上采集全眼,并将其送去进行分子和形态学表型分析。在这篇视频文章中,我们展示了摘除并转移未固定和灌注固定的小鼠眼睛以进行远程表型分析的技术。

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