Working with Xenopus spinal neurons in live cell culture

Timothy M. Gómez, Dan Harrigan, John Henley, Estuardo Robles

Research output: Contribution to journalReview article

32 Scopus citations

Abstract

Neurons from the Xenopus spinal cord are highly versatile and easily manipulated, making them an ideal model system to answer questions regarding the cellular and molecular basis of early neural development and function. Xenopus has been a productive model system in studies ranging from axon growth and guidance to synaptic plasticity. Exogenous molecules, such as proteins, fluorescent tracers, and nucleic acids, can be injected into early blastomeres to load tracers in all neurons or into late blastomeres to target specifc classes of neurons based on established lineage maps. Xenopus spinal neurons also provide an excellent culture system, as neurons extend processes on a variety of substrata and develop at room temperature in minimal salt solutions. Live fluorescent neurons can be imaged for hours with fluorescence microscopy at room temperature in static cultures without neurotrophic support or serum. This highly reduced culture system minimizes variables that can confound interpretation of results. Cultures can be prepared at various stages of development as dissociated neurons or as spinal cord explants. Both excitatory and inhibitory neurons develop in culture, and synaptic contacts among neurons and between neurons and nonneuronal targets form naturally. The simple anatomy and rapid rostral-to-caudal development of the Xenopus spinal cord also make this an excellent in vivo model system to analyze axon guidance by identifiable classes of neurons. This chapter focuses on techniques that exploit both in vitro and in vivo qualities of this system.

Original languageEnglish (US)
Pages (from-to)129-156
Number of pages28
JournalMethods in cell biology
Volume2003
Issue number71
DOIs
StatePublished - 2003

ASJC Scopus subject areas

  • Cell Biology

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    Gómez, T. M., Harrigan, D., Henley, J., & Robles, E. (2003). Working with Xenopus spinal neurons in live cell culture. Methods in cell biology, 2003(71), 129-156. https://doi.org/10.1016/s0091-679x(03)01008-2