Visualization of newly synthesized neuronal RNA in vitro and in vivo using click-chemistry

利用点击化学技术在体内和体外可视化新合成的神经元 RNA

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作者:Güney Akbalik, Kasper Langebeck-Jensen, Georgi Tushev, Sivakumar Sambandan, Jennifer Rinne, Irina Epstein, Iván Cajigas, Irena Vlatkovic, Erin M Schuman

Abstract

The neuronal transcriptome changes dynamically to adapt to stimuli from the extracellular and intracellular environment. In this study, we adapted for the first time a click chemistry technique to label the newly synthesized RNA in cultured hippocampal neurons and intact larval zebrafish brain. Ethynyl uridine (EU) was incorporated into neuronal RNA in a time- and concentration-dependent manner. Newly synthesized RNA granules observed throughout the dendrites were colocalized with mRNA and rRNA markers. In zebrafish larvae, the application of EU to the swim water resulted in uptake and labeling throughout the brain. Using a GABA receptor antagonist, PTZ (pentylenetetrazol), to elevate neuronal activity, we demonstrate that newly transcribed RNA signal increased in specific regions involved in neurogenesis.

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