MicroRNA Profiling of Alzheimer’s Disease Cerebrospinal Fluid

  • Johannes Denk
  • Holger JahnEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1750)


MicroRNAs (miRNAs) are a class of small, highly conserved, and noncoding RNAs that modulate gene expression by regulating the activity and stability of target mRNAs. MiRNAs play significant roles by controlling fundamental cellular processes and its deregulation is associated with various diseases. Ubiquitous expression and its release into circulation make them interesting biomarkers, which can be measured by different platforms. In this book chapter, we provide a specific protocol that describes the detection of circulating miRNAs in CSF by using RT-qPCR.

Key words

RT-qPCR CSF MicroRNA MIQE Biomarker SYBR-green CNS disorders 


  1. 1.
    Weber JA, Baxter DH, Zhang S et al (2010) The microRNA spectrum in 12 body fluids. Clin Chem 56(11):1733–1741. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Mitchell PS, Parkin RK, Kroh EM et al (2008) Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A 105(30):10513–10518. CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Balusu S, Van Wonterghem E, De Rycke R et al (2016) Identification of a novel mechanism of blood-brain communication during peripheral inflammation via choroid plexus-derived extracellular vesicles. EMBO Mol Med 8(10):1162–1183. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Pritchard CC, Cheng HH, Tewari M (2012) MicroRNA profiling: approaches and considerations. Nat Rev Genet 13(5):358–369. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Mestdagh P, Hartmann N, Baeriswyl L et al (2014) Evaluation of quantitative miRNA expression platforms in the microRNA quality control (miRQC) study. Nat Methods 11(8):809–815. CrossRefPubMedGoogle Scholar
  6. 6.
    Witwer KW, Halushka MK (2016) Toward the promise of microRNAs—enhancing reproducibility and rigor in microRNA research. RNA Biol 13(11):1103–1116. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4):611–622. CrossRefPubMedGoogle Scholar
  8. 8.
    Bustin SA (2010) Why the need for qPCR publication guidelines? The case for MIQE. Methods 50(4):217–226. CrossRefPubMedGoogle Scholar
  9. 9.
    Bustin SA, Beaulieu JF, Huggett J et al (2010) MIQE précis: practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments. BMC Mol Biol 11:74. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Kirschner MB, van Zandwijk N, Reid G (2013) Cell-free microRNAs: potential biomarkers in need of standardized reporting. Front Genet 4:56. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Kirschner MB, Edelman JJB, Kao SCH et al (2013) The impact of hemolysis on cell-free microRNA biomarkers. Front Genet 4:94., Article No.: 94CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Mestdagh P, Van Vlierberghe P, De Weer A et al (2009) A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol 10(6):R64. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    D’Haene B, Mestdagh P, Hellemans J et al (2012) miRNA expression profiling: from reference genes to global mean normalization. Methods Mol Biol 822:261–272. CrossRefPubMedGoogle Scholar
  14. 14.
    Schwarzenbach H, da Silva AM, Calin G et al (2015) Data normalization strategies for MicroRNA quantification. Clin Chem 61(11):1333–1342. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Broeders S, Huber I, Grohmann L et al (2014) Guidelines for validation of qualitative real-time PCR methods. Trends Food Sci Tech 37(2):115–126. CrossRefGoogle Scholar
  16. 16.
    Kralik P, Ricchi M (2017) A basic guide to real time PCR in microbial diagnostics: definitions, parameters, and everything. Front Microbiol 8:108. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Vandesompele J, De Preter K, Pattyn F et al (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):Research0034CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Andersen CL, Jensen JL, Orntoft TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64(15):5245–5250. CrossRefPubMedGoogle Scholar
  19. 19.
    El-Khoury V, Pierson S, Kaoma T et al (2016) Assessing cellular and circulating miRNA recovery: the impact of the RNA isolation method and the quantity of input material. Sci Rep 6:19529. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    McAlexander MA, Phillips MJ, Witwer KW (2013) Comparison of methods for miRNA extraction from plasma and quantitative recovery of RNA from cerebrospinal fluid. Front Genet 4:83. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Department of Psychiatry and PsychotherapyUniversity Medical Center Hamburg-EppendorfHamburgGermany

Personalised recommendations