Unit

UNIT 2D.15 Revealing Neuronal Circuitry Using Stem Cell-Derived Neurons

  1. Isabella Garcia1,2,
  2. Cynthia Kim2,3,
  3. Benjamin R. Arenkiel1,3,4

Published Online: 1 MAY 2013

DOI: 10.1002/9780470151808.sc02d15s25

Current Protocols in Stem Cell Biology

Current Protocols in Stem Cell Biology

How to Cite

Garcia, I., Kim, C. and Arenkiel, B. R. 2013. Revealing Neuronal Circuitry Using Stem Cell-Derived Neurons. Current Protocols in Stem Cell Biology. 25:D:2D.15:2D.15.1–2D.15.18.

Author Information

  1. 1

    Program in Developmental Biology, Baylor College of Medicine, Houston, Texas

  2. 2

    Medical Scientist Training Program, Baylor College of Medicine, Houston, Texas

  3. 3

    Department of Molecular & Human Genetics, Baylor College of Medicine, Houston, Texas

  4. 4

    Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, Houston, Texas

Publication History

  1. Published Online: 1 MAY 2013

Abstract

Mouse embryonic stem cell (mESC)-derived neurons are a renewable cell source for investigation of neuronal circuits. Engineering circuit-tracing components into stem cells facilitates studies on mechanisms of synaptic coupling and circuitogenesis. This unit details methods for the generation of mESC-derived neurons harboring trans-synaptic viral tracing elements, which are used for investigation of synaptic connections within circuits in vitro, ex vivo, and in vivo. The first protocol describes procedures for feeder-free passaging of mESCs, modified to carry reporter and rabies virus tracing elements. The second protocol describes in vitro generation of neurons from these ESCs. The last protocols describe the use of ESC-derived neurons as “source cells” for rabies virus circuit-tracing to identify inputs onto synaptically connected neurons. Given the broad applicability, these protocols can be applied to investigate the ability of in vitro-derived neurons to establish/maintain synaptic connections in disease models, and/or with human-induced pluripotent stem cells. Curr. Protoc. Stem Cell Biol. 25:2D.15.1-2D.15.18. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • monosynaptic;
  • embryonic stem cells;
  • retrograde;
  • rabies;
  • circuit;
  • neuron;
  • synapse;
  • TVA;
  • rabies-G