Postsynaptic scaffolds of excitatory and inhibitory synapses in hippocampal neurons: Maintenance of core components independent of actin filaments and microtubules

Daniel W. Allison*, Adam S. Chervin, Vladimir I. Gelfand, Ann Marie Craig

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

190 Scopus citations

Abstract

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 Ca2+/calmodulin-dependent protein kinase II (CaMKII)α, another core component of the PSD, was completely dependent on an intact actin cytoskeleton and was partially disrupted by detergent. Drebrin and α- 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.

Original languageEnglish (US)
Pages (from-to)4545-4554
Number of pages10
JournalJournal of Neuroscience
Volume20
Issue number12
DOIs
StatePublished - Jun 15 2000

Keywords

  • Actin
  • CaMKIIα
  • Drebrin
  • GABA receptor
  • GKAP
  • Gephyrin
  • Microtubules
  • PSD- 95
  • Postsynaptic density
  • α-actinin-2

ASJC Scopus subject areas

  • Neuroscience(all)

Fingerprint Dive into the research topics of 'Postsynaptic scaffolds of excitatory and inhibitory synapses in hippocampal neurons: Maintenance of core components independent of actin filaments and microtubules'. Together they form a unique fingerprint.

Cite this