ArticleFrontiers in cellular neuroscience2013
Combined fluorescent in situ hybridization for detection of microRNAs and immunofluorescent labeling for cell-type markers.
Article in Frontiers in cellular neuroscience, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed.
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- Subcellular Transcriptomics and Proteomics: A Comparative Methods Review.Molecular & cellular proteomics : MCP · 2022Review
- Inhibition of microRNA-494-3p activates Wnt signaling and reduces proinflammatory macrophage polarization in atherosclerosis.Molecular therapy. Nucleic acids · 2021Article
- A comprehensive study to delineate the role of an extracellular vesicle-associated microRNA-29a in chronic methamphetamine use disorder.Journal of extracellular vesicles · 2021Article
- Expression of miR-200c corresponds with increased reactive oxygen species and hypoxia markers after transient focal ischemia in mice.Neurochemistry international · 2021Article
- Epigenetic down-regulation of microRNA-126 in scleroderma endothelial cells is associated with impaired responses to VEGF and defective angiogenesis.Journal of cellular and molecular medicine · 2021Article
- Article
- A novel approach for microRNA in situ hybridization using locked nucleic acid probes.Scientific reports · 2021Article
- How to distinguish between different cell lineages sharing common markers using combinations of double in-situ-hybridization and immunostaining in avian embryos: CXCR4-positive mesodermal and neural crest-derived cells.Histochemistry and cell biology · 2021Article
- Identification of ncRNA-Mediated Functions of Nucleus-Localized miR-320 in Cardiomyocytes.Molecular therapy. Nucleic acids · 2020Article
- circDLPAG4/HECTD1 mediates ischaemia/reperfusion injury in endothelial cells via ER stress.RNA biology · 2020Article
- Pre-treatment with microRNA-181a Antagomir Prevents Loss of Parvalbumin Expression and Preserves Novel Object Recognition Following Mild Traumatic Brain Injury.Neuromolecular medicine · 2019Article
- In the Telencephalon, GluN2C NMDA Receptor Subunit mRNA is Predominately Expressed in Glial Cells and GluN2D mRNA in Interneurons.Neurochemical research · 2019Article
- Introduction to Digital Image Analysis in Whole-slide Imaging: A White Paper from the Digital Pathology Association.Journal of pathology informatics · 2019Review
- Astrocyte EV-Induced lincRNA-Cox2 Regulates Microglial Phagocytosis: Implications for Morphine-Mediated Neurodegeneration.Molecular therapy. Nucleic acids · 2018Article
- Digital Microscopy, Image Analysis, and Virtual Slide Repository.ILAR journal · 2018Article
- Anti-miRNA oligonucleotides: A comprehensive guide for design.RNA biology · 2018Review
- Tat-Mediated Induction of miRs-34a & -138 Promotes Astrocytic Activation via Downregulation of SIRT1: Implications for Aging in HAND.Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology · 2017Article
- Recent advances in high-performance fluorescent and bioluminescent RNA imaging probes.Chemical Society reviews · 2017Review
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Authors and funding
3 authors.
Funding
Abstract
Identification of the cell type of origin for normal or aberrant gene expression is critical for many studies, and poses a significant problem for some regulatory RNAs such as microRNAs. MicroRNAs are small non-coding RNAs that regulate cellular function by targeting specific mRNAs and reducing the level of their protein product. Aberrant expression of miRNAs in cell-types where they are not normally expressed occurs in several disease conditions. Therefore, it is important to determine not only the expression level of microRNAs, but also where they are expressed. Here we describe a detailed method for fluorescent in situ hybridization (FISH) combined with immunofluorescent labeling for cell-type markers in formalin fixed paraffin embedded (FFPE) sections along with modifications required to adapt the protocol for primary neurons grown in culture. We have combined the specificity and stability of locked nucleic acid (LNA) probes with tyramide signal amplification. To prevent loss of small RNA species, we performed post-fixation with ethylcarbodiimide (EDC). Additionally by omitting protease digestion and using only high temperature with sodium citrate buffer for FFPE sections, we were able to perform immunolabeling for proteins concurrently with in situ hybridization without compromising efficacy of either procedure.
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