全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
PLOS ONE  2013 

Acute Inactivation of PSD-95 Destabilizes AMPA Receptors at Hippocampal Synapses

DOI: 10.1371/journal.pone.0053965

Full-Text   Cite this paper   Add to My Lib

Abstract:

Postsynatptic density protein (PSD-95) is a 95 kDa scaffolding protein that assembles signaling complexes at synapses. Over-expression of PSD-95 in primary hippocampal neurons selectively increases synaptic localization of AMPA receptors; however, mice lacking PSD-95 display grossly normal glutamatergic transmission in hippocampus. To further study the scaffolding role of PSD-95 at excitatory synapses, we generated a recombinant PSD-95-4c containing a tetracysteine motif, which specifically binds a fluorescein derivative and allows for acute and permanent inactivation of PSD-95. Interestingly, acute inactivation of PSD-95 in rat hippocampal cultures rapidly reduced surface AMPA receptor immunostaining, but did not affected NMDA or transferrin receptor localization. Acute photoinactivation of PSD-95 in dissociated neurons causes ~80% decrease in GluR2 surface staining observed by live-cell microscopy within 15 minutes of PSD-95-4c ablation. These results confirm that PSD-95 stabilizes AMPA receptors at postsynaptic sites and provides insight into the dynamic interplay between PSD-95 and AMPA receptors in live neurons.

References

[1]  Funke L, Dakoji S, Bredt DS (2005) Membrane-associated guanylate kinases regulate adhesion and plasticity at cell junctions. Annual Review of Biochemistry 74: 219–245.
[2]  Kim E, Sheng M (2004) PDZ domain proteins of synapses. Nature Reviews Neuroscience 5: 771–781.
[3]  Sheng M, Sala C (2001) PDZ domains and the organization of supramolecular complexes. Annual Review of Neuroscience 24: 1–29.
[4]  Opazo P, Sainlos M, Choquet D (2011) Regulation of AMPA receptor surface diffusion by PSD-95 slots. Current opinion in neurobiology. doi:10.1016/j.conb.2011.10.010.
[5]  Tochio H, Mok YK, Zhang Q, Kan HM, Bredt DS, et al. (2000) Formation of nNOS/PSD-95 PDZ dimer requires a preformed beta-finger structure from the nNOS PDZ domain. Journal of Molecular Biology 303: 359–370.
[6]  Olsen O, Bredt DS (2003) Functional analysis of the nucleotide binding domain of membrane-associated guanylate kinases. The Journal of Biological Chemistry 278: 6873–6878.
[7]  Bredt DS, Nicoll R a (2003) AMPA receptor trafficking at excitatory synapses. Neuron 40: 361–379.
[8]  El-Husseini AE, Schnell E, Chetkovich DM, Nicoll RA, Bredt DS (2000) PSD-95 involvement in maturation of excitatory synapses. Science 290: 1364–1368.
[9]  Schnell E, Sizemore M, Karimzadegan S, Chen L, Bredt DS, et al. (2002) Direct interactions between PSD-95 and stargazin control synaptic AMPA receptor number. Proceedings of the National Academy of Sciences of the United States of America 99: 13902–13907.
[10]  Stein V, House DRC, Bredt DS, Nicoll RA (2003) Postsynaptic density-95 mimics and occludes hippocampal long-term potentiation and enhances long-term depression. Journal of Neuroscience 23: 5503–5506.
[11]  Jackson AC, Nicoll RA (2011) The expanding social network of ionotropic glutamate receptors: TARPs and other transmembrane auxiliary subunits. Neuron 70: 178–199.
[12]  Tomita S, Adesnik H, Sekiguchi M, Zhang W, Wada K, et al. (2005) Stargazin modulates AMPA receptor gating and trafficking by distinct domains. Nature 435: 1052–1058.
[13]  Chen L, Chetkovich DM, Petralia RS, Sweeney NT, Kawasaki Y, et al. (2000) Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms. Nature 408: 936–943.
[14]  Bats C, Groc L, Choquet D (2007) The interaction between Stargazin and PSD-95 regulates AMPA receptor surface trafficking. Neuron 53: 719–734.
[15]  Sainlos M, Tigaret C, Poujol C, Olivier NB, Bard L, et al. (2011) Biomimetic divalent ligands for the acute disruption of synaptic AMPAR stabilization. Nature chemical biology 7: 81–91.
[16]  Griffin BA, Adams SR, Tsien RY (1998) Specific covalent labeling of recombinant protein molecules inside live cells. Science 281: 269–272.
[17]  Marek KW, Davis GW (2002) Transgenically encoded protein photoinactivation (FlAsH-FALI): acute inactivation of synaptotagmin I. Neuron. 36: 805–813.
[18]  Habets RLP, Verstreken P (2011) FlAsH-FALI Inactivation of a Protein at the Third-Instar Neuromuscular Junction. Cold Spring Harbor Protocols 2011: pdb.prot5597–pdb.prot5597.
[19]  Beck S, Sakurai T, Eustace BK, Beste G, Schier R, et al. (2002) Fluorophore-assisted light inactivation: a high-throughput tool for direct target validation of proteins. Proteomics 2: 247–255.
[20]  Habets RLP, Verstreken P (2011) Construction and Expression of Tetracysteine-Tagged Proteins for FlAsH-FALI. Cold Spring Harbor Protocols 2011: pdb.prot5596–pdb.prot5596.
[21]  El-Husseini AE-D, Schnell E, Dakoji S, Sweeney N, Zhou Q, et al. (2002) Synaptic strength regulated by palmitate cycling on PSD-95. Cell 108: 849–863.
[22]  DiCiommo DP, Bremner R (1998) Rapid, high level protein production using DNA-based Semliki Forest virus vectors. The Journal of Biological Chemistry 273: 18060–18066.
[23]  Craven SE, El-Husseini AE, Bredt DS (1999) Synaptic targeting of the postsynaptic density protein PSD-95 mediated by lipid and protein motifs. Neuron 22: 497–509.
[24]  Brewer GJ, Torricelli JR, Evege EK, Price PJ (1993) Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. Journal of Neuroscience Research 35: 567–576.
[25]  El-Husseini AE, Craven SE, Chetkovich DM, Firestein BL, Schnell E, et al. (2000) Dual palmitoylation of PSD-95 mediates its vesiculotubular sorting, postsynaptic targeting, and ion channel clustering. Journal Of Cell Biology 148: 3030–3035.
[26]  El-Husseini AE-D, Schnell E, Dakoji S, Sweeney N, Zhou Q, et al. (2002) Synaptic Strength Regulated by Palmitate Cycling on PSD-95. Cell 108: 849–863.
[27]  Bredt DS, Nicoll RA (2003) AMPA Receptor Trafficking at Excitatory Synapses. Neuron 40: 361–379.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133