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

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.