Detection of mammalian microRNA expression by in situ hybridization with RNA oligonucleotides

We have developed an in situ hybridization procedure for the detection of microRNAs (miRNAs) in tissue sections from mouse embryos and adult organs. The method uses highly specific washing conditions for RNA oligonucleotide probes conjugated to a fluorescein hapten. We show that this method detects...

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Bibliographic Details
Published in:Developmental dynamics 2006-09, Vol.235 (9), p.2538-2548
Main Authors: Deo, Monika, Yu, Jenn‐Yah, Chung, Kwan‐Ho, Tippens, Melissa, Turner, David L.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
brain
Brain - growth & development
Brain - metabolism
Cell Line
Central Nervous System - embryology
Central Nervous System - metabolism
choroid plexus
Embryonic Development - genetics
Eye - growth & development
Eye - metabolism
Female
Fluorescein
Gene Expression
In Situ Hybridization
Mice
microRNA
MicroRNAs - genetics
Oligonucleotide Probes - genetics
Oligoribonucleotides - genetics
Pregnancy
retina
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Summary:We have developed an in situ hybridization procedure for the detection of microRNAs (miRNAs) in tissue sections from mouse embryos and adult organs. The method uses highly specific washing conditions for RNA oligonucleotide probes conjugated to a fluorescein hapten. We show that this method detects predominantly mature miRNAs rather than the miRNA precursors or primary transcripts. We have determined expression patterns for several miRNAs expressed in the developing and adult nervous system, including miR‐124a, miR‐9, miR‐92, and miR‐204. Whereas miR‐124a is expressed in neurons, miR‐9 is expressed in neural progenitors and some neurons, and miR‐204 is expressed in the choroid plexus, retinal pigment epithelium, and ciliary body. miR‐204 is located in an intron of the TRPM3 gene, and the TRPM3 mRNA is coexpressed with miR‐204 in the choroid plexus. We also find that primary transcripts for miR‐124a and miR‐9 genes are expressed in patterns similar to their respective mature miRNAs. The ability to visualize expression of specific miRNAs in embryos and tissues should aid studies on miRNA function. Developemental Dynamics 235:2538–2548, 2006. © 2006 Wiley‐Liss, Inc.
ISSN:1058-8388
1097-0177
DOI:10.1002/dvdy.20847