Real-Time Quantitative PCR and Fluorescence In Situ Hybridization for Subcellular Localization of miRNAs in Neurons

Antonis Tatarakis; Danesh Moazed

doi:10.1007/978-1-0716-1916-2_1

Real-Time Quantitative PCR and Fluorescence In Situ Hybridization for Subcellular Localization of miRNAs in Neurons

Methods Mol Biol. 2022:2417:1-17. doi: 10.1007/978-1-0716-1916-2_1.

Authors

Antonis Tatarakis¹, Danesh Moazed²

Affiliations

¹ Department of Cell Biology, and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA. antonis_tatarakis@hms.harvard.edu.
² Department of Cell Biology, and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA. danesh@hms.harvard.edu.

PMID: 35099787
DOI: 10.1007/978-1-0716-1916-2_1

Abstract

Neuronal miRNAs play major roles in regulation of synaptic development and plasticity. The small size of miRNAs and, in some cases, their low level of expression make their quantification and detection challenging. Here, we outline methods to quantify steady state levels of miRNAs in neurons and the brain by using real-time quantitative PCR (RT-qPCR) and to determine miRNA subcellular localization in primary neurons by a sensitive fluorescence in situ hybridization (FISH) method.

Keywords: Brain; Expression profile; Isolation; Neurons; miRNA RT-qPCR assays; miRNA fluorescence in situ hybridization; miRNAs.

MeSH terms

In Situ Hybridization, Fluorescence
MicroRNAs* / metabolism
Neurons / metabolism
Real-Time Polymerase Chain Reaction / methods

Substances

MicroRNAs