斑马鱼幼体和成体牙齿组织的全组织免疫组化和原位杂交

Whole mount immunohistochemistry and in situ hybridization of larval and adult zebrafish dental tissues.

作者信息

Verstraeten Barbara, Sanders Ellen, Huysseune Ann

机构信息

Evolutionary Developmental Biology, Department of Biology, Ghent University, Ghent, Belgium.

出版信息

Methods Mol Biol. 2012;887:179-91. doi: 10.1007/978-1-61779-860-3_16.

DOI:10.1007/978-1-61779-860-3_16

PMID:22566056

Abstract

Tooth development is increasingly being studied in a variety of vertebrate model organisms, each contributing its own perspective to our understanding of dental diversity. In situ hybridization and immunohistochemistry are well-established and frequently used techniques to study the presence of mRNA and protein. Here, we describe a protocol for whole mount immunohistochemistry and in situ hybridization that can be applied to all stages of zebrafish development and dissected bony parts. The description of these protocols is followed by the outline of a quick decalcification method and the procedure for embedding in epoxy resin to obtain serial sections with high histological quality.

摘要

牙齿发育越来越多地在各种脊椎动物模型生物中进行研究，每种生物都为我们理解牙齿多样性提供了自己的视角。原位杂交和免疫组织化学是研究mRNA和蛋白质存在的成熟且常用的技术。在这里，我们描述了一种用于整体免疫组织化学和原位杂交的方案，该方案可应用于斑马鱼发育的所有阶段以及解剖后的骨骼部分。在这些方案的描述之后，是一种快速脱钙方法的概述以及环氧树脂包埋的程序，以获得具有高组织学质量的连续切片。