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SRF binding to SRE 6.9 in the Arc promoter is essential for LTD in cultured Purkinje cells

Nature Neuroscience volume 13, pages 1082–1089 (2010)Cite this article

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Abstract

It has been suggested that gene expression and protein synthesis are required for both long-term memory consolidation and late phases of long-term potentiation and long-term depression (LTD). The necessary genes and the specific transcription factor binding sites in their promoters remain unknown. We found that inhibition of the transcription factor SRF or its cofactor MAL blocked the late phase of LTD in mouse cultured cerebellar Purkinje cells, as did deletion of the immediate early gene Arc. Using neuronal bacterial artificial chromosome (BAC) transfection, we found that, in Arc−/− cells transfected with a wild-type Arc BAC, late-phase LTD was rescued. However, mutation of one SRF-binding site in the Arc promoter (SRE 6.9) blocked this rescue. Co-transfection of wild-type Arc and SRF engineered to bind mutated SRE 6.9 restored late-phase LTD in Arc−/−, SRE 6.9 mutant BAC cells. Thus, SRF binding to SRE 6.9 in the Arc promoter is required for the late phase of cerebellar LTD.

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Figure 1: Interfering with SRF blocks the late phase of cerebellar LTD in cultured Purkinje cells.
Figure 2: The late phase of cerebellar LTD is blocked in Arc−/− cultured Purkinje cells.
Figure 3: Expression of a BAC construct containing Arc and its genomic regulatory regions rescues early- and late-phase LTD in Arc−/− neurons.
Figure 4: The failure of the SRE 6.9 mutant Arc-BAC to rescue late-phase LTD is reversed by co-transfection with a plasmid encoding a Gal4–SRF fusion protein.
Figure 5: Manipulations that inhibit actin-MAL signaling block the late phase of cerebellar LTD in cultured Purkinje cells.

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Acknowledgements

We thank D. VanNess for excellent technical assistance and to members of the Linden and Worley laboratories for helpful suggestions. M. Greenberg's laboratory (Harvard University) provided the Gal4-SRF fusion construct plasmid. The authors thank B. Knöll (University of Tübingen) for sharing β-actin constructs and for helpful suggestions. The FLAG-tagged dominant-negative MAL was a generous gift from R. Prywes (Columbia University). This work supported by grants from the US Public Health Service (NIH R37 MH51106 to D.J.L., P50 MH084020 to D.J.L., D.D.G., P.F.W. and R.L.H., and NIH 5K12 NS001696-07 to C.S.-H.), the National Basic Research Program of China (2004CB518800 to B.X. and 2004CB518800 to X.Z.) and the Science Fund for Creative Research Group, China (B.X.).

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Author notes

  1. Jason D Shepherd

    Present address: Present address: The Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.,

Authors and Affiliations

  1. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Constance Smith-Hicks, Bo Xiao, Jason D Shepherd, Richard L Huganir, David D Ginty, Paul F Worley & David J Linden

  2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Constance Smith-Hicks & Paul F Worley

  3. The State Key Laboratory of Biotherapy, West-China Hospital, Sichuan University, Chengdu, China

    Bo Xiao, Rongkang Deng, Yifei Ji & Xia Zhao

  4. Howard Hughes Medical Institute, USA

    Jason D Shepherd, Richard L Huganir & David D Ginty

  5. Department of Molecular Biology, Max Planck Institute of Biochemistry, Martinsried, Germany

    Guido Posern

  6. Institute for Molecular and Cellular Cognition, Center for Molecular Neurobiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Dietmar Kuhl

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Contributions

C.S.-H. performed the immunohistochemistry and designed a portion of the study. B.X., R.D., Y.J. and X.Z. engineered the BACs. J.D.S. performed immunohistochemistry and wrote an early draft of the manuscript. G.P. provided mutant actin reagents and helped to design the experiments that used them. D.K. provided Arc−/− mice. R.L.H., D.D.G. and P.F.W. supervised the project. D.J.L. provided overall supervision of the project and conducted the electrophysiological and imaging experiments.

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Correspondence to David J Linden.

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Smith-Hicks, C., Xiao, B., Deng, R. et al. SRF binding to SRE 6.9 in the Arc promoter is essential for LTD in cultured Purkinje cells. Nat Neurosci 13, 1082–1089 (2010). https://doi.org/10.1038/nn.2611

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