Culturing conditions determine neuronal and glial excitability

Sandra Stoppelkamp, Gernot Riedel, Bettina Platt

Research output: Contribution to journalArticle

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Abstract

The cultivation of pure neuronal cultures is considered advantageous for the investigation of cell-type specific responses (such as transmitter release and also pharmacological agents), however, divergent results are a likely consequence of media modifications and culture composition. Using Fura-2 based imaging techniques, we here set out to compare calcium responses of rat hippocampal neurones and glia to excitatory stimulation with l-glutamate in different culture types and media. Neurones in neurone-enriched cultures had increased responses to 10 µM and 100 µM l-glutamate (+43 and 45%, respectively; p's < 0.001) and a slower recovery compared to mixed cultures, indicating heightened excitability. In matured (15–20 days in vitro) mixed cultures, neuronal responder rates were suppressed in a neurone-supportive medium (Neurobasal-A, NB: 65%) compared to a general-purpose medium (supplemented minimal essential medium, MEM: 96%). Glial response size in contrast did not differ greatly in isolated or mixed cultures maintained in MEM, but responder rates were suppressed in both culture types in NB (e.g. 10 µM l-glutamate responders in mixed cultures: 29% in NB, 71% in MEM). This indicates that medium composition is more important for glial excitability than the presence of neurones, whereas the presence of glia has an important impact on neuronal excitability. Therefore, careful consideration of culturing conditions is crucial for interpretation and comparison of experimental results. Especially for investigations of toxicity and neuroprotection mixed cultures may be more physiologically relevant over isolated cultures as they comprise aspects of mutual influences between glia and neurones.
Original languageEnglish
Pages (from-to)132-138
Number of pages7
JournalJournal of Neuroscience Methods
Volume194
Issue number1
Early online date15 Oct 2010
DOIs
Publication statusPublished - 15 Dec 2010

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Neuroglia
Neurons
Glutamic Acid
Culture Media
Fura-2
Pharmacology
Calcium

Keywords

  • calcium homeostasis
  • l-Glutamate
  • Fura-2
  • neurobasal
  • MEM
  • rat
  • hippocampus

Cite this

Culturing conditions determine neuronal and glial excitability. / Stoppelkamp, Sandra; Riedel, Gernot; Platt, Bettina.

In: Journal of Neuroscience Methods, Vol. 194, No. 1, 15.12.2010, p. 132-138.

Research output: Contribution to journalArticle

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