Decoding Arc Transcription: a Live-cell Study of Stimulation Patterns and Transcriptional Output

Overview

Journal Learn Mem

Specialty Neurology

Date 2024 Sep 11

PMID 39260877

Authors

Dong Wook Kim

Hyungseok C Moon

Byung Hun Lee

Hye Yoon Park

Affiliations

Soon will be listed here.

Abstract

Activity-regulated cytoskeleton-associated protein (Arc) plays a crucial role in synaptic plasticity, a process integral to learning and memory. Arc transcription is induced within a few minutes of stimulation, making it a useful marker for neuronal activity. However, the specific neuronal activity patterns that initiate Arc transcription have remained elusive due to the inability to observe mRNA transcription in live cells in real time. Using a genetically encoded RNA indicator (GERI) mouse model that expresses endogenous Arc mRNA tagged with multiple GFPs, we investigated Arc transcriptional activity in response to various electrical field stimulation patterns. The GERI mouse model was generated by crossing the Arc-PBS knock-in mouse, engineered with binding sites in the 3' untranslated region (UTR) of Arc mRNA, and the transgenic mouse expressing the cognate binding protein fused to GFP. In dissociated hippocampal neurons, we found that the pattern of stimulation significantly affects Arc transcription. Specifically, theta-burst stimulation consisting of high-frequency (100 Hz) bursts delivered at 10 Hz frequency induced the highest rate of Arc transcription. Concurrently, the amplitudes of nuclear calcium transients also reached their peak with 10 Hz burst stimulation, indicating a correlation between calcium concentration and transcription. However, our dual-color single-cell imaging revealed that there were no significant differences in calcium amplitudes between Arc-positive and Arc-negative neurons upon 10 Hz burst stimulation, suggesting the involvement of other factors in the induction of Arc transcription. Our live-cell RNA imaging provides a deeper insight into the complex regulation of transcription by activity patterns and calcium signaling pathways.

Citing Articles

Brain Plasticity and Cell Competition: Immediate Early Genes Are the Focus.

Tregub P, Komleva Y, Kukla M, Averchuk A, Vetchinova A, Rozanova N Cells. 2025; 14(2).

PMID: 39851571 PMC: 11763428. DOI: 10.3390/cells14020143.

References

Yilmaz-Rastoder E, Miyamae T, Braun A, Thiels E . LTP- and LTD-inducing stimulations cause opposite changes in arc/arg3.1 mRNA level in hippocampal area CA1 in vivo. Hippocampus. 2010; 21(12):1290-301. PMC: 3006082. DOI: 10.1002/hipo.20838. View

West A, Chen W, Dalva M, Dolmetsch R, Kornhauser J, Shaywitz A . Calcium regulation of neuronal gene expression. Proc Natl Acad Sci U S A. 2001; 98(20):11024-31. PMC: 58677. DOI: 10.1073/pnas.191352298. View

Nonaka M, Kim R, Fukushima H, Sasaki K, Suzuki K, Okamura M . Region-specific activation of CRTC1-CREB signaling mediates long-term fear memory. Neuron. 2014; 84(1):92-106. DOI: 10.1016/j.neuron.2014.08.049. View

Vazdarjanova A, McNaughton B, Barnes C, Worley P, Guzowski J . Experience-dependent coincident expression of the effector immediate-early genes arc and Homer 1a in hippocampal and neocortical neuronal networks. J Neurosci. 2002; 22(23):10067-71. PMC: 6758761. View

Guzowski J, Miyashita T, Chawla M, Sanderson J, Maes L, Houston F . Recent behavioral history modifies coupling between cell activity and Arc gene transcription in hippocampal CA1 neurons. Proc Natl Acad Sci U S A. 2006; 103(4):1077-82. PMC: 1347968. DOI: 10.1073/pnas.0505519103. View

Gartner A, Staiger V . Neurotrophin secretion from hippocampal neurons evoked by long-term-potentiation-inducing electrical stimulation patterns. Proc Natl Acad Sci U S A. 2002; 99(9):6386-91. PMC: 122958. DOI: 10.1073/pnas.092129699. View

Jiang Y, VanDongen A . Selective increase of correlated activity in Arc-positive neurons after chemically induced long-term potentiation in cultured hippocampal neurons. eNeuro. 2021; . PMC: 8658543. DOI: 10.1523/ENEURO.0540-20.2021. View

Kawashima T, Okuno H, Nonaka M, Adachi-Morishima A, Kyo N, Okamura M . Synaptic activity-responsive element in the Arc/Arg3.1 promoter essential for synapse-to-nucleus signaling in activated neurons. Proc Natl Acad Sci U S A. 2009; 106(1):316-21. PMC: 2629236. DOI: 10.1073/pnas.0806518106. View

Buzsaki G, Logothetis N, Singer W . Scaling brain size, keeping timing: evolutionary preservation of brain rhythms. Neuron. 2013; 80(3):751-64. PMC: 4009705. DOI: 10.1016/j.neuron.2013.10.002. View

10.

Miyashita T, Kubik S, Haghighi N, Steward O, Guzowski J . Rapid activation of plasticity-associated gene transcription in hippocampal neurons provides a mechanism for encoding of one-trial experience. J Neurosci. 2009; 29(4):898-906. PMC: 2749324. DOI: 10.1523/JNEUROSCI.4588-08.2009. View

11.

Waltereit R, Dammermann B, Wulff P, Scafidi J, Staubli U, Kauselmann G . Arg3.1/Arc mRNA induction by Ca2+ and cAMP requires protein kinase A and mitogen-activated protein kinase/extracellular regulated kinase activation. J Neurosci. 2001; 21(15):5484-93. PMC: 6762636. View

12.

Pulimood N, Contreras M, Pruitt M, Tarasiewicz A, Medina A . Phosphorylation of CREB at Serine 142 and 143 Is Essential for Visual Cortex Plasticity. eNeuro. 2021; 8(5). PMC: 8555886. DOI: 10.1523/ENEURO.0217-21.2021. View

13.

Vera M, Biswas J, Senecal A, Singer R, Park H . Single-Cell and Single-Molecule Analysis of Gene Expression Regulation. Annu Rev Genet. 2016; 50:267-291. PMC: 5149423. DOI: 10.1146/annurev-genet-120215-034854. View

14.

Balkowiec A, Katz D . Cellular mechanisms regulating activity-dependent release of native brain-derived neurotrophic factor from hippocampal neurons. J Neurosci. 2002; 22(23):10399-407. PMC: 6758764. View

15.

Colgan L, Parra-Bueno P, Holman H, Tu X, Jain A, Calubag M . Dual Regulation of Spine-Specific and Synapse-to-Nucleus Signaling by PKCδ during Plasticity. J Neurosci. 2023; 43(30):5432-5447. PMC: 10376934. DOI: 10.1523/JNEUROSCI.0208-22.2023. View

16.

Guzowski J, McNaughton B, Barnes C, Worley P . Environment-specific expression of the immediate-early gene Arc in hippocampal neuronal ensembles. Nat Neurosci. 1999; 2(12):1120-4. DOI: 10.1038/16046. View

17.

Lyford G, Yamagata K, Kaufmann W, Barnes C, Sanders L, Copeland N . Arc, a growth factor and activity-regulated gene, encodes a novel cytoskeleton-associated protein that is enriched in neuronal dendrites. Neuron. 1995; 14(2):433-45. DOI: 10.1016/0896-6273(95)90299-6. View

18.

Peebles C, Yoo J, Thwin M, Palop J, Noebels J, Finkbeiner S . Arc regulates spine morphology and maintains network stability in vivo. Proc Natl Acad Sci U S A. 2010; 107(42):18173-8. PMC: 2964216. DOI: 10.1073/pnas.1006546107. View

19.

Mayr B, Montminy M . Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol. 2001; 2(8):599-609. DOI: 10.1038/35085068. View

20.

Gao M, Sossa K, Song L, Errington L, Cummings L, Hwang H . A specific requirement of Arc/Arg3.1 for visual experience-induced homeostatic synaptic plasticity in mouse primary visual cortex. J Neurosci. 2010; 30(21):7168-78. PMC: 2881313. DOI: 10.1523/JNEUROSCI.1067-10.2010. View