单足动物胚胎的全载原位杂交。

CSH protocols Pub Date : 2008-10-01 DOI:10.1101/pdb.prot5076
Anna L Keyte, Kathleen K Smith
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引用次数: 5

摘要

家鼠,灰色短尾或实验室负鼠,是最常用的实验室有袋动物。除了使它成为一种方便的实验动物(体型小,易于护理,非季节性繁殖)的因素外,它还是第一个基因组被测序的有袋动物。Monodelphis已被证明是一种有用的模式生物,用于研究脊髓再生,紫外线(UV)诱导的黑色素瘤,遗传对胆固醇的影响,以及免疫系统的比较研究。此外,Monodelphis已被用来了解嗅觉系统的基本功能和各种嗅觉化学物质在社会和生殖行为中的作用。最近,Monodelphis已被用于了解有袋动物发展的基本方面,解剖学,进化,以及衍生的有袋动物发展和繁殖模式的进化后果。本协议详细说明了单足动物胚胎的全安装原位杂交,但它广泛适用于任何有袋动物。有袋动物胚胎的各个阶段的特殊条件都包括在整个方案中。然而,由于角质层的形成阻止了探针和抗体的渗透,整个早产儿胚胎阶段(~ 33期至出生)已被证明很难进行研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Whole-mount in situ hybridization in monodelphis embryos.

INTRODUCTIONMonodelphis domestica, the gray, short-tailed, or laboratory opossum, is the most commonly used laboratory marsupial. In addition to the factors that make it a convenient laboratory animal (small size, ease of care, nonseasonal breeding), it is the first marsupial whose genome has been sequenced. Monodelphis has proven useful as a model organism for studies on spinal cord regeneration, ultraviolet (UV)-induced melanoma, and genetic influences on cholesterol, as well as comparative studies of the immune system. In addition, Monodelphis has been used to understand the basic functions of the olfactory system and the role of various olfactory chemicals in social and reproductive behavior. Recently, Monodelphis has been used to understand fundamental aspects of marsupial development, anatomy, evolution, and evolutionary consequences of the derived marsupial mode of development and reproduction. This protocol details whole-mount in situ hybridization of Monodelphis embryos, but it is broadly applicable to any marsupial. Special conditions have been included throughout the protocol for various stages of marsupial embryos. Nevertheless, whole, preterm embryonic stages (~stage 33 to birth) have proven to be difficult to work with because formation of the cuticle prevents probe and antibody penetration.

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