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In Situ Detection of Neuron-Specific MicroRNAs in Frozen Brain Tissue

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Epigenetic Methods in Neuroscience Research

Part of the book series: Neuromethods ((NM,volume 105))

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Abstract

MicroRNAs (miRNAs) are 22–24 nucleotide long small RNA molecules, which regulate the expression of protein coding genes by binding to their 3′-untranslated region and causing inhibition of protein synthesis either by degradation of the target mRNA or by repression of translation. The vast majority of microRNAs are found in the brain, yet their functions are still to be investigated. Therefore, being able to analyze the spatial and the temporal expression of the miRNAs at the tissue and cellular levels in the brain is of greatest interest. Due to small size of miRNAs and low expression levels, it has been a challenge to detect miRNAs in situ. Here, we describe a fluorescence in situ hybridization (FISH) method based on locked nucleic acid (LNA) probes and the tyramide signal amplification (TSA) system for detection of microRNAs in frozen brain tissue sections. Combining this miRNA-FISH method with immunofluorescence using neuron-specific antibodies allows cell type-specific localization of miRNAs in the brain.

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Acknowledgements

This research was supported by grants from the Swedish Research Council, Funds from Karolinska Institutet, Åhlén-stiftelsen, and The Lundbeck Foundation. We thank Mattias Karlén for help with the schematic drawings.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3, 2200, Copenhagen, Denmark

    Asli Silahtaroglu

  2. Department of Cellular and Molecular Pathology, German Cancer Research Center, Heidelberg, Germany

    Silke Herzer

  3. Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden

    Silke Herzer & Björn Meister

Authors
  1. Asli Silahtaroglu
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  2. Silke Herzer
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  3. Björn Meister
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Corresponding author

Correspondence to Asli Silahtaroglu .

Editor information

Editors and Affiliations

  1. Neuroscience Center, University of Helsinki, Helsinki, Finland

    Nina Karpova

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Silahtaroglu, A., Herzer, S., Meister, B. (2016). In Situ Detection of Neuron-Specific MicroRNAs in Frozen Brain Tissue. In: Karpova, N. (eds) Epigenetic Methods in Neuroscience Research. Neuromethods, vol 105. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2754-8_13

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  • DOI: https://doi.org/10.1007/978-1-4939-2754-8_13

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2753-1

  • Online ISBN: 978-1-4939-2754-8

  • eBook Packages: Springer Protocols

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