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对新生小鼠视网膜进行全层免疫荧光染色,以研究体内血管生成。

Whole mount immunofluorescent staining of the neonatal mouse retina to investigate angiogenesis in vivo.

作者信息

Tual-Chalot Simon, Allinson Kathleen R, Fruttiger Marcus, Arthur Helen M

机构信息

Institute of Genetic Medicine, Newcastle University, Australia.

出版信息

J Vis Exp. 2013 Jul 9(77):e50546. doi: 10.3791/50546.

Abstract

Angiogenesis is the complex process of new blood vessel formation defined by the sprouting of new blood vessels from a pre-existing vessel network. Angiogenesis plays a key role not only in normal development of organs and tissues, but also in many diseases in which blood vessel formation is dysregulated, such as cancer, blindness and ischemic diseases. In adult life, blood vessels are generally quiescent so angiogenesis is an important target for novel drug development to try and regulate new vessel formation specifically in disease. In order to better understand angiogenesis and to develop appropriate strategies to regulate it, models are required that accurately reflect the different biological steps that are involved. The mouse neonatal retina provides an excellent model of angiogenesis because arteries, veins and capillaries develop to form a vascular plexus during the first week after birth. This model also has the advantage of having a two-dimensional (2D) structure making analysis straightforward compared with the complex 3D anatomy of other vascular networks. By analyzing the retinal vascular plexus at different times after birth, it is possible to observe the various stages of angiogenesis under the microscope. This article demonstrates a straightforward procedure for analyzing the vasculature of a mouse retina using fluorescent staining with isolectin and vascular specific antibodies.

摘要

血管生成是一个复杂的新血管形成过程,其定义为从预先存在的血管网络中萌发出新的血管。血管生成不仅在器官和组织的正常发育中起关键作用,而且在许多血管形成失调的疾病中也起关键作用,如癌症、失明和缺血性疾病。在成年期,血管通常处于静止状态,因此血管生成是新型药物开发的一个重要靶点,旨在尝试在疾病中特异性地调节新血管的形成。为了更好地理解血管生成并制定适当的调节策略,需要能够准确反映所涉及的不同生物学步骤的模型。小鼠新生视网膜提供了一个极好的血管生成模型,因为动脉、静脉和毛细血管在出生后的第一周内发育形成一个血管丛。与其他血管网络复杂的三维解剖结构相比,该模型还具有二维(2D)结构的优势,使得分析变得简单直接。通过在出生后的不同时间分析视网膜血管丛,可以在显微镜下观察血管生成的各个阶段。本文展示了一种使用异凝集素和血管特异性抗体进行荧光染色来分析小鼠视网膜脉管系统的简单方法。

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