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Postsynaptic Scaffolds of Excitatory and Inhibitory Synapses in Hippocampal Neurons: Maintenance of Core Components Independent of Actin Filaments and Microtubules
Open accessAbstract
The mechanisms responsible for anchoring molecular components of postsynaptic specializations in the mammalian brain are not well understood but are presumed to involve associations with cytoskeletal elements. Here we build on previous studies of neurotransmitter receptors (Allison et al., 1998) to analyze the modes of attachment of scaffolding and signal transducing proteins of both glutamate and GABA postsynaptic sites to either the microtubule or microfilament cytoskeleton. Hippocampal pyramidal neurons in culture were treated with latrunculin A to depolymerize actin, with vincristine to depolymerize microtubules, or with Triton X-100 to extract soluble proteins. The synaptic clustering of PSD-95, a putative NMDA receptor anchoring protein and a core component of the postsynaptic density (PSD), was unaffected by actin depolymerization, microtubule depolymerization, or detergent extraction. The same was largely true for GKAP, a PSD-95-interacting protein. In contrast, the synaptic clustering of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII)alpha, another core component of the PSD, was completely dependent on an intact actin cytoskeleton and was partially disrupted by detergent. Drebrin and alpha-actinin-2, actin-binding proteins concentrated in spines, were also dependent on F-actin for synaptic localization but were unaffected by detergent extraction. Surprisingly, the subcellular distributions of the inhibitory synaptic proteins GABA(A)R and gephyrin, which has a tubulin-binding motif, were unaffected by depolymerization of microtubules or actin or by detergent extraction. These studies reveal an unsuspected heterogeneity in the modes of attachment of postsynaptic proteins to the cytoskeleton and support the idea that PSD-95 and gephyrin may be core scaffolding components independent of the actin or tubulin cytoskeleton.
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“Lastly, the actin cytoskeleton is known to mediate receptor positioning (Gudheti et al., 2013) and its depolymerization was shown to reduce glutamate receptor accumulations in spines (Allison et al., 2000).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Allison et al., 2000
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Metadata
- Authors
- Allison, D. W., Chervin, A. S., Gelfand, V. I., & Craig, A. M.
- Affiliations
- University of Illinois Urbana-Champaign (US)
- Venue
- The Journal of Neuroscience
- Publisher
- Society for Neuroscience
- Published
- 2000-06-15
- Volume
- 20
- Issue
- 12
- Pages
- 4545-4554
- ISSN
- 0270-6474 (ISSN-L)
- 1529-2401
- PMC
- PMC6772458
- OpenAlex
- W2148256023
- Cited by
- 245
- References
- 79
- Citations by year
- 2025: 5 · 2024: 3 · 2023: 2 · 2022: 4 · 2021: 10 · 2020: 4 · 2019: 3 · 2018: 4 · 2017: 5 · 2016: 2 · 2015: 5 · 2014: 12 · 2013: 8 · 2012: 7
- FWCI
- 8.28
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation
- National Institutes of Health (GM-52111)
- National Institute of General Medical Sciences (R01GM052111, GM52111-01)
- National Institute of Neurological Disorders and Stroke (NS33184, R01NS033184)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetics and Neurodevelopmental Disorders
- Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
- Keywords
- postsynaptic scaffolds, postsynaptic density, actin cytoskeleton, microtubules, PSD-95, GKAP, CaMKIIα, drebrin, ACTN2, GABAA receptor, gephyrin, glutamatergic synapses, inhibitory synapses, hippocampal pyramidal neurons, latrunculin A, vincristine, detergent extraction, dendritic spines
- MeSH terms
- Disks Large Homolog 4 Protein, Actins, Animals, Cells, Cultured, Evoked Potentials, Hippocampus, Marine Toxins, Membrane Proteins, Microtubules, Nerve Tissue Proteins, Neurons, Synapses, Thiazoles, Vincristine, Octoxynol, Bridged Bicyclo Compounds, Heterocyclic, Excitatory Postsynaptic Potentials, Intracellular Signaling Peptides and Proteins, Rats, Thiazolidines
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