Co-culture Synaptogenic Assay: A New Look at Fluorescence Reporters and Technological Devices

Methods Mol Biol. 2017;1538:13-27. doi: 10.1007/978-1-4939-6688-2_2.

Abstract

The mechanism underlying the differentiation of pre- and postsynaptic specifications involves the sequential and dynamic recruitment of specific molecules coordinated by bidirectional signaling across the synaptic cleft. In this chapter, we describe the co-culture assay, a useful method to evaluate cell-surface molecules through its ability to promote the recruitment of proteins required for synapse structure and function. The versatility of this simple and reliable method is illustrated by the wide variety of applications ranging from analysis of synaptogenic activity to evaluation of soluble compounds with therapeutic potential. In addition, we provide a framework to enable the co-culture assay as a tool for high-throughput studies, thereby improving the efficiency and sensitivity of this classic method in neuroscience.

Keywords: Adhesion molecules; Co-culture assay; Confocal imaging; High throughput; Synaptogenesis.

MeSH terms

  • Animals
  • Biomarkers
  • Cell Adhesion Molecules / metabolism
  • Cell Line
  • Coculture Techniques*
  • Drug Discovery / methods
  • Gene Expression
  • Genes, Reporter
  • High-Throughput Screening Assays
  • Hippocampus / cytology
  • Hippocampus / physiology
  • Humans
  • Immunohistochemistry
  • Microscopy, Confocal / methods
  • Neurons / cytology
  • Neurons / physiology
  • Small Molecule Libraries
  • Synapses / drug effects
  • Synapses / physiology*
  • Synaptic Transmission*

Substances

  • Biomarkers
  • Cell Adhesion Molecules
  • Small Molecule Libraries