Real-time imaging of Arc/Arg3.1 transcription ex vivo reveals input-specific immediate early gene dynamics

Abstract

The ability of neurons to process and store salient environmental features underlies information processing in the brain. Long-term information storage requires synaptic plasticity and regulation of gene expression. While distinct patterns of activity have been linked to synaptic plasticity, their impact on immediate early gene (IEG) expression remains poorly understood. The activity regulated cytoskeleton associated (Arc) gene has received wide attention as an IEG implicated in synaptic plasticity and memory. Yet, to date, the transcriptional dynamics of Arc in response to compartment and input-specific activity is unclear. By developing a knock-in mouse to fluorescently tag Arc alleles, we studied real-time transcription dynamics after stimulation of dentate granule cells (GCs) in acute hippocampal slices. To our surprise, we found that Arc transcription displayed distinct temporal kinetics depending on the activation of excitatory inputs that convey functionally distinct information, i.e. medial and lateral perforant paths (MPP and LPP, respectively). Moreover, the transcriptional dynamics of Arc after synaptic stimulation was similar to direct activation of GCs, although the contribution of ionotropic glutamate receptors, L-type voltage gated calcium channel, and the endoplasmic reticulum (ER) differed. Specifically, we observed an ER-mediated synapse-to-nucleus signal that supported elevations in nuclear calcium, and rapid induction of Arc transcription following MPP stimulation. However, activation of LPP inputs displayed lower nuclear calcium rise, which could underlie the delayed transcriptional onset of Arc. Our findings highlight how input-specific activity distinctly impacts transcriptional dynamics of an IEG linked to learning and memory. Significance statementEnvironmental experiences trigger neuronal activity that elicits gene expression in the nervous system. Rapid induction of specific genes known as immediate early genes (IEGs) supports activity-dependent changes of neuronal circuits to ultimately influence animal behavior. However, the cellular and molecular mechanisms controlling how distinct forms of neuronal activity modulate IEG expression remains unclear. The activity regulated cytoskeleton associated (Arc) gene is a critical IEG linked to memory. By imaging Arc transcription in real-time after neuronal activity, we identified how different receptors and signaling pathways influence transcriptional induction and dynamics of an IEG. Our findings provide insights into how information received by distinct synaptic inputs could be encoded by modulating IEG dynamics.