Unit

UNIT 26.4 Compartmented Neuron Cultures for Directional Infection by Alpha Herpesviruses

  1. Dušica Curanović1,
  2. Toh Hean Ch'ng1,2,
  3. Moriah Szpara1,
  4. Lynn Enquist1

Published Online: 1 JUN 2009

DOI: 10.1002/0471143030.cb2604s43

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Curanović, D., Ch'ng, T. H., Szpara, M. and Enquist, L. 2009. Compartmented Neuron Cultures for Directional Infection by Alpha Herpesviruses. Current Protocols in Cell Biology. 43:26.4:26.4.1–26.4.23.

Author Information

  1. 1

    Department of Molecular Biology, Princeton University, Princeton, New Jersey

  2. 2

    University of California at Los Angeles, Los Angeles, California

Publication History

  1. Published Online: 1 JUN 2009
  2. Published Print: JUN 2009

Abstract

Compartmented neuronal cultures allow experimenters to establish separate fluid environments for neuronal axons and the soma from which they emanate. Physical isolation of cell bodies and axons is achieved by culturing neurons in tri-chambered Teflon rings. Dissociated ganglia are plated in one end compartment of the trichamber, and axonal growth is guided underneath watertight silicone grease barriers into a separate compartment. Since the axons and cell bodies are located in different compartments, they can be infected and assayed separately. We describe the assembly and use of compartmented neuronal cultures for in vitro study of directional infection of neurons by alpha herpesviruses. Selective application of viral inoculum to only one compartment ensures that the remainder of the neuron is not contaminated by input inoculum. This allows for quantification of viral spread, and unambiguous interpretation of immunofluorescence and electron microscopy images. Curr. Protoc. Cell Biol. 43:26.4.1-26.4.23. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • compartmented neuron culture;
  • Campenot chambers;
  • tri-chamber;
  • anterograde;
  • retrograde;
  • alpha herpesvirus;
  • virus