title: Visualization of newly synthesized neuronal RNA in vitro and in vivo using click-chemistry.
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.
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Güney
authors: G
authors: Akbalik
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Kasper
authors: K
authors: Langebeck-Jensen
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Georgi
authors: G
authors: Tushev
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Sivakumar
authors: S
authors: Sambandan
authors: b Institute for Organic Chemistry and Chemical Biology, Goethe University , Frankfurt , Germany.
authors: Jennifer
authors: J
authors: Rinne
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Irina
authors: I
authors: Epstein
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Iván
authors: I
authors: Cajigas
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Irena
authors: I
authors: Vlatkovic
authors: a Max Planck Institute for Brain Research , Frankfurt , Germany.
authors: Erin M
authors: EM
authors: Schuman
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D057930:Click Chemistry; D020459:Genes, rRNA; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism*; D039102:Poly(A)-Binding Proteins / Q000378:metabolism; D017966:Pyramidal Cells / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish
chemical_list: D039102:Poly(A)-Binding Proteins; D012333:RNA, Messenger; D012313:RNA
keywords: Nascent RNA; PABP; RNA in zebrafish brain; Y10B; neuronal RNA; newly synthesized RNA; visualization of RNA