The Exocyst Component Exo70 Modulates Dendrite Arbor Formation, Synapse Density, and Spine Maturation in Primary Hippocampal Neurons

Mol Neurobiol. 2019 Jul;56(7):4620-4638. doi: 10.1007/s12035-018-1378-0. Epub 2018 Oct 29.

Abstract

Neurons are highly polarized cells displaying an elaborate architectural morphology. The design of their dendritic arborization and the distribution of their synapses contribute importantly to information processing in the brain. The growth and complexity of dendritic arbors are driven by the formation of synapses along their lengths. Synaptogenesis is augmented by the secretion of factors, like BDNF, Reelin, BMPs, or Wnts. Exo70 is a component of the exocyst complex, a protein complex that guides membrane addition and polarized exocytosis. While it has been linked to cytokinesis and the establishment of cell polarity, its role in synaptogenesis is poorly understood. In this report, we show that Exo70 plays a role in the arborization of dendrites and the development of synaptic connections between cultured hippocampal neurons. Specifically, while the overexpression of Exo70 increases dendritic arborization, synapse number, and spine density, the inhibition of Exo70 expression reduces secondary and tertiary dendrite formation and lowers synapse density. Moreover, increasing Exo70 expression augmented synaptic vesicle recycling as evaluated by FM4-64 dye uptake and the inverse was observed with downregulation of endogenous Exo70. Monitoring the formation of dendritic spines by super-resolution microscopy, we also observed that mRFP-Exo70 accumulates at the tip of EGFP-β-actin-positive filopodia. Together, these results suggest that Exo70 is essentially involved in the formation of synapses and neuronal dendritic morphology.

Keywords: Dendrite; Dendritic spine; Exo70; Exocyst; Neuron; Synapse.

MeSH terms

  • Animals
  • Cells, Cultured
  • Dendritic Spines / metabolism*
  • Down-Regulation / genetics
  • HEK293 Cells
  • Hippocampus / metabolism*
  • Humans
  • Lentivirus / metabolism
  • Models, Biological
  • Phenotype
  • Rats, Sprague-Dawley
  • Synapses / metabolism*
  • Vesicular Transport Proteins / metabolism*

Substances

  • Exoc7 protein, rat
  • Vesicular Transport Proteins