PUBLICATION
Observation of miRNA Gene Expression in Zebrafish Embryos by In Situ Hybridization to MicroRNA Primary Transcripts
- Authors
- He, X., Yan, Y.L., Delaurier, A., and Postlethwait, J.H.
- ID
- ZDB-PUB-110207-25
- Date
- 2011
- Source
- Zebrafish 8(1): 1-8 (Journal)
- Registered Authors
- DeLaurier, April, He, Xinjun, Postlethwait, John H., Yan, Yi-Lin
- Keywords
- none
- MeSH Terms
-
- MicroRNAs/chemistry
- MicroRNAs/genetics*
- MicroRNAs/metabolism
- Digoxigenin/chemistry
- RNA Processing, Post-Transcriptional
- DNA, Intergenic
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- Gene Expression Regulation, Developmental
- Animals
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- SOX9 Transcription Factor/genetics
- SOX9 Transcription Factor/metabolism
- Embryo, Nonmammalian/metabolism
- In Situ Hybridization/methods*
- PubMed
- 21288128 Full text @ Zebrafish
Citation
He, X., Yan, Y.L., Delaurier, A., and Postlethwait, J.H. (2011) Observation of miRNA Gene Expression in Zebrafish Embryos by In Situ Hybridization to MicroRNA Primary Transcripts. Zebrafish. 8(1):1-8.
Abstract
MicroRNAs (miRNAs) add a previously unexpected layer to the post-transcriptional regulation of protein production. Although locked nucleic acids (LNAs) reveal the distribution of mature miRNAs by in situ hybridization (ISH) experiments in zebrafish and other organisms, high cost has restricted their use. Further, LNA probes designed to recognize mature miRNAs do not distinguish expression patterns of two miRNA genes that produce the same mature miRNA sequence. Riboprobes are substantially less expensive than LNAs, but have not been used to detect miRNA gene expression because they do not bind with high affinity to the short, 22-nucleotide-long mature miRNAs. To solve these problems, we capitalized on the fact that miRNAs are initially transcribed into long primary transcripts (pri-mRNAs). We show here that conventional digoxigenin-labeled riboprobes can bind to primary miRNA transcripts in zebrafish embryos. We tested intergenic and intronic miRNAs (miR-10d, miR-21, miR-27a, miR-126a, miR-126b, miR-138, miR-140, miR-144, miR-196a1, miR-196a2, miR-196a2b [miR-196c], miR-196b, miR-196b1b [miR-196d], miR-199, miR-214, miR-200, and miR-222) in whole mounts and some of these in histological sections. Results showed that pri-miRNA ISH provides an attractive and cost-effective tool to study miRNA expression by ISH. We use this method to show that miR-126a and miR-126b are transcribed in the caudal vasculature in the pattern of their neighboring gene ci116 or host gene egfl7, respectively, and that the chondrocyte miRNA mir-140 lies downstream of Sox9 in development of the craniofacial skeleton.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping