A transgenic mouse for imaging activity-dependent dynamics of endogenous arc mRNA in live neurons

Sulagna Das, Hyungseok C. Moon, Robert H. Singer, Hye Yoon Park

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

Localized translation plays a crucial role in synaptic plasticity and memory consolidation. However, it has not been possible to follow the dynamics of memory-associated mRNAs in living neurons in response to neuronal activity in real time. We have generated a novel mouse model where the endogenous Arc/Arg3.1 gene is tagged in its 3′ untranslated region with stem-loops that bind a bacteriophage PP7 coat protein (PCP), allowing visualization of individual mRNAs in real time. The physiological response of the tagged gene to neuronal activity is identical to endogenous Arc and reports the true dynamics of Arc mRNA from transcription to degradation. The transcription dynamics of Arc in cultured hippocampal neurons revealed two novel results: (i) A robust transcriptional burst with prolonged ON state occurs after stimulation, and (ii) transcription cycles continue even after initial stimulation is removed. The correlation of stimulation with Arc transcription and mRNA transport in individual neurons revealed that stimulus-induced Ca2+ activity was necessary but not sufficient for triggering Arc transcription and that blocking neuronal activity did not affect the dendritic transport of newly synthesized Arc mRNAs. This mouse will provide an important reagent to investigate how individual neurons transduce activity into spatiotemporal regulation of gene expression at the synapse.

Original languageEnglish (US)
Article numbereaar3448
JournalScience Advances
Volume4
Issue number6
DOIs
StatePublished - Jun 20 2018
Externally publishedYes

Bibliographical note

Funding Information:
We thank C. Guo and Janelia Gene Targeting and Transgenics Resources for cloning, electroporation, ES cell screening, and microinjection. We also thank A. Senecal for help with analysis of transcription sites from live imaging. For carrying out the behavioral experiments, we thank M. Gulinello from Behavioral Core, Albert Einstein College of Medicine (Bronx, NY). We thank C. Nwokafor and S. H. Kim for animal maintenance and M. L. Jones for genotyping. This research was supported by NIH grant NS083085 to R.H.S. and Samsung Science and Technology Foundation project number SSTF-BA1602-11 to H.Y.P.

Publisher Copyright:
Copyright © 2018 The Authors.

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