Protocol

Part of the series Neuromethods pp 1-7

Date:

Measuring miRNA Mediated Translational Regulation with Live Cell Imaging

  • Min Jeong KyeAffiliated withInstitute of Human Genetics, University of Cologne Email author 

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Abstract

Numerous microRNAs are detected in synaptic areas such as axons and dendrites. As accurate regulation of local protein synthesis can be crucial for the function of neurons, miRNAs can play very important roles for mRNA translation in the rather isolated cellular spaces such as synaptic area. However, due to the technical limitation, it is very difficult to measure efficiency of protein synthesis in the synaptic area with biochemical methods. Therefore, visualizing translation and measuring protein levels at the synaptic sites by imaging techniques can be a good alternative. Fluorescence recovery after photobleaching (FRAP) has been widely used to measure local protein synthesis rate in axons. This technique allows us to measure speed and efficiency of translation of certain mRNAs. We modified this method to measure how miRNAs influence on local synthesis of proteins in neurons.

Keywords:

MicroRNA Local translation Axon FRAP (fluorescence recovery after photobleaching) Live cell imaging