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揭示细节：斑马鱼胚胎和成年组织的全胚胎 miRNA 原位杂交方案。

Revealing details: whole mount microRNA in situ hybridization protocol for zebrafish embryos and adult tissues.

机构信息

Hubrecht Institute-KNAW ; Utrecht University Medical Center.

出版信息

Biol Open. 2012 Jun 15;1(6):566-9. doi: 10.1242/bio.2012810. Epub 2012 Apr 11.

DOI:10.1242/bio.2012810
PMID:23213449
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3509442/
Abstract

Non-coding microRNA (miRNA) molecules bind their target mRNAs and thereby modulate the amount of protein produced. To understand the significance of a potential miRNA-mRNA interaction, temporal and spatial information on miRNA and mRNA expression is essential. Here, we provide a detailed protocol for miRNA whole mount in situ hybridization. We introduce the use of Morpholino based oligos as antisense probes for miRNA detection, in addition to the current "gold standard" locked nucleic acid (LNA) probes. Furthermore we have modified existing miRNA in situ protocols thereby improving both sensitivity and resolution of miRNA visualization in whole zebrafish embryos and adult tissues.

摘要

非编码 microRNA（miRNA）分子与其靶 mRNAs 结合，从而调节蛋白质的产生量。为了理解潜在的 miRNA-mRNA 相互作用的意义，miRNA 和 mRNA 表达的时空信息是必不可少的。在这里，我们提供了 miRNA 整体原位杂交的详细方案。我们引入了基于 Morpholino 的寡核苷酸作为 miRNA 检测的反义探针，除了当前的“金标准”锁核酸（LNA）探针。此外，我们还修改了现有的 miRNA 原位方案，从而提高了整个斑马鱼胚胎和成年组织中 miRNA 可视化的灵敏度和分辨率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/b4a814b2a512/bio-01-06-566-f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/da9ed8fa6619/bio-01-06-566-f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/9c86640917b2/bio-01-06-566-f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/b4a814b2a512/bio-01-06-566-f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/da9ed8fa6619/bio-01-06-566-f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/9c86640917b2/bio-01-06-566-f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/07e3/3509442/b4a814b2a512/bio-01-06-566-f03.jpg

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