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发育中的真皮淋巴管的组织学和形态学特征

Histological and Morphological Characterization of Developing Dermal Lymphatic Vessels.

作者信息

Betterman Kelly L, Harvey Natasha L

机构信息

Centre for Cancer Biology, University of South Australia and SA Pathology, Adelaide, SA, Australia.

出版信息

Methods Mol Biol. 2018;1846:19-35. doi: 10.1007/978-1-4939-8712-2_2.

Abstract

The capacity to visualize the lymphatic vasculature in three-dimensions has revolutionized our understanding of the morphogenetic mechanisms important for constructing the lymphatic vascular network during development. Two complementary approaches are commonly employed to assess the function of genes and signaling pathways important for development of the dermal lymphatic vasculature in the mouse embryo. The first of these is whole-mount immunostaining of embryonic skin to analyze dermal lymphatic vessel network patterning and morphology in two and three dimensions. The second is immunostaining of thin tissue sections to examine lymphatic vessel identity, lumen formation and protein localization within discrete lymphatic endothelial cells in a two-dimensional setting. Here we present detailed protocols for multicolor immunofluorescent immunostaining of embryonic dorsal skin and thin tissue cryosections. Each of these methods generates high-resolution images of the dermal lymphatic vasculature, yielding information integral to in-depth characterization of lymphatic vessel phenotypes in the developing mouse embryo.

摘要

在三维空间中可视化淋巴管系统的能力彻底改变了我们对发育过程中构建淋巴管网络至关重要的形态发生机制的理解。通常采用两种互补方法来评估对小鼠胚胎真皮淋巴管系统发育重要的基因和信号通路的功能。其中第一种是对胚胎皮肤进行整体免疫染色,以在二维和三维空间中分析真皮淋巴管网络的模式和形态。第二种是对薄组织切片进行免疫染色,以在二维环境中检查离散淋巴管内皮细胞内的淋巴管身份、管腔形成和蛋白质定位。在这里,我们展示了胚胎背部皮肤和薄组织冰冻切片的多色免疫荧光免疫染色的详细方案。这些方法中的每一种都能生成真皮淋巴管系统的高分辨率图像,为深入表征发育中小鼠胚胎的淋巴管表型提供不可或缺的信息。

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