Neuronal cell cultures from Aplysia for high-resolution imaging of growth cones

Aih Cheun Lee, Boris Decourt, Daniel M. Suter

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Neuronal growth cones are the highly motile structures at the tip of axons that can detect guidance cues in the environment and transduce this information into directional movement towards the appropriate target cell. To fully understand how guidance information is transmitted from the cell surface to the underlying dynamic cytoskeletal networks, one needs a model system suitable for live cell imaging of protein dynamics at high temporal and spatial resolution. Typical vertebrate growth cones are too small to quantitatively analyze F-actin and microtubule dynamics. Neurons from the sea hare Aplysia californica are 5-10 times larger than vertebrate neurons, can easily be kept at room temperature and are very robust cells for micromanipulation and biophysical measurements. Their growth cones have very defined cytoplasmic regions and a well-described cytoskeletal system. The neuronal cell bodies can be microinjected with a variety of probes for studying growth cone motility and guidance. In the present protocol we demonstrate a procedure for dissection of the abdominal ganglion, culture of bag cell neurons and setting up an imaging chamber for live cell imaging of growth cones.

Original languageEnglish (US)
Article numbere662
JournalJournal of Visualized Experiments
Issue number12
DOIs
StatePublished - Feb 2008

Keywords

  • Abdominal ganglion
  • Aplysia californica
  • Bag cell neuron
  • Cytoskeletal dynamics
  • Growth cone motility and guidance
  • Issue 12
  • Live cell imaging
  • Nervous system
  • Neuronal cell culture
  • Neuronal growth cone
  • Neuroscience

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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