Detection of Mammalian Microrna Expression by in Situ Hybridization with Rna Oligonucleotides.
From: Molecular and Behavioral Neuroscience Institute, University of Michigan, Ann Arbor, Michigan 48109-2200, USA.
Developmental dynamics : an official publication of the American Association of Anatomists
- Publish Date: Sep 2006
- ISSN: 1058-8388
- Volume: 235
- Issue: 9
- Pages: 2538-48
- Medium: Print
- Language: English
- Citation (JAMA): Deo Monika, Yu Jenn-Yah, Chung Kwan-Ho, et al. Detection of Mammalian Microrna Expression by in Situ Hybridization with Rna Oligonucleotides.. Dev. Dyn. Sep 2006;235:2538-48
Abstract
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.
Mesh Headings (Keywords): Animals, Base Sequence, Brain, Cell Line, Central Nervous System, Embryonic Development, Eye, Female, Fluorescein, Gene Expression, In Situ Hybridization, Mice, MicroRNAs, Oligonucleotide Probes, Oligoribonucleotides, Pregnancy
Check for Full Text / PubMed Unique Identifier (PMID): 16736490
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