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Activity-dependent regulation of dendritic synthesis and trafficking of AMPA receptors

Nature Neuroscience volume 7, pages 244–253 (2004)Cite this article

Abstract

Regulation of AMPA receptor (AMPAR) trafficking is important for neural plasticity. Here we examined the trafficking and synthesis of the GluR1 and GluR2 subunits using ReAsH-EDT2 and FlAsH-EDT2 staining. Activity blockade of rat cultured neurons increased dendritic GluR1, but not GluR2, levels. Examination of transected dendrites revealed that both AMPAR subunits were synthesized in dendrites and that activity blockade enhanced dendritic synthesis of GluR1 but not GluR2. In contrast, acute pharmacological manipulations increased dendritic synthesis of both subunits. AMPARs synthesized in dendrites were inserted into synaptic plasma membranes and, after activity blockade, the electrophysiological properties of native synaptic AMPARs changed in the manner predicted by the imaging experiments. In addition to providing a novel mechanism for synaptic modifications, these results point out the advantages of using FlAsH-EDT2 and ReAsH-EDT2 for studying the trafficking of newly synthesized proteins in local cellular compartments such as dendrites.

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Figure 1: ReAsH-EDT2 and FlAsH-EDT2 are not toxic and specifically stain tetracysteine-tagged GluR1 and GluR2.
Figure 2: Characterization of basal AMPAR subunit trafficking.
Figure 3: Chronic activity blockade increased dendritic levels, but not the rate of transport, of GluR1.
Figure 4: Local synthesis of AMPAR subunits occurs in isolated dendrites.
Figure 5: Chronic activity blockade enhances local dendritic synthesis of GluR1.
Figure 6: A decrease in lysosome-mediated degradation does not account for the increase in GluR1 following activity blockade.
Figure 7: Acute treatments increase local dendritic synthesis of both GluR1 and GluR2.
Figure 8: Chronic activity blockade enhances synaptic localization of GluR1 in isolated dendrites and changes the subunit composition of native synaptic AMPARs.

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Acknowledgements

We thank J. Fisher and E. Saura for technical assistance and members of the Malenka and Garner labs for suggestions. cDNAs were gifts from M. Sheng (AMPAR subunits), Z. Jia (AMPAR subunits), H. Schulman (α and βCaMKII) and M. Mayford (α and βCaMKII). This work was supported by grants from the National Institutes of Health (R.C.M., MH63394; R.Y.T., NS27177; M.E., P41-RR04050) and Howard Hughes Medical Institute (R.Y.T.).

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Authors and Affiliations

  1. Department of Psychiatry and Behavioral Sciences, Nancy Friend Pritzker Laboratory, 1201 Welch Road, Stanford University School of Medicine, Palo Alto, 94304, California, USA

    William Ju, Wade Morishita, Jennifer Tsui, Craig C Garner & Robert C Malenka

  2. Department of Neurosciences 0608, National Center for Microscopy & Imaging Research, University of California San Diego, 9500 Gilman Drive, La Jolla, 92093, California, USA

    Guido Gaietta, Thomas J Deerinck & Mark H Ellisman

  3. Department of Pharmacology, Howard Hughes Medical Institute, University of California San Diego, 9500 Gilman Drive, La Jolla, 92093, California, USA

    Stephen R Adams & Roger Y Tsien

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Correspondence to Robert C Malenka.

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Competing interests

R.Y. Tsien is co-inventor on the patents for FlAsH and ReAsH, which the University of California has licensed, and will receive a share of any royalties that the University of California collects. He has no financial relationship with Invitrogen, the company that is now commercializing these reagents.

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Ju, W., Morishita, W., Tsui, J. et al. Activity-dependent regulation of dendritic synthesis and trafficking of AMPA receptors. Nat Neurosci 7, 244–253 (2004). https://doi.org/10.1038/nn1189

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