Long-term in vitro maintenance of neuromuscular junction activity of Drosophila larvae

Ryan Ball, Bin Xing, Philip Bonner, Joseph Shearer, Robin L. Cooper

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The larval Drosophila neuromuscular junction (NMJ) has proven to be an excellent system to test fundamental aspects of synaptic transmission, such as relationships among ion channel function, subtypes of glutamate receptors, and the functions of synaptic proteins in the presynaptic compartment. Recent advances in understanding bi-directional communication between nerves and muscles of Drosophila are helping uncover developmental as well as maintenance cues that could be applicable to all chemical synapses. The development of HL3 medium makes it possible to record synaptic responses at NMJs for prolonged periods of time. We demonstrate that media commonly used to culture CNS neurons and imaginal disks of Drosophila such as Schneider's and M3 completely block glutamatergic synaptic transmission at the NMJ. The depressed postsynaptic excitatory junction potentials (EJPs) partially recover from exposure to such media shortly after switching to the HL3 medium. Preliminary results from NMJs of filleted 3rd instar larvae for 4 days in vitro bathed in a modified HL3 medium show great promise. The resting membrane potential and the EJP amplitudes after 4 days in vitro are normal. These results demonstrate the possibility for chronic studies of developmental regulation in culture, which in some cases are impractical in the whole animal.

Original languageEnglish
Pages (from-to)247-255
Number of pages9
JournalComparative Biochemistry and Physiology - A Molecular and Integrative Physiology
Volume134
Issue number2
DOIs
StatePublished - Feb 1 2003

Bibliographical note

Funding Information:
Funding was provided by a University of Kentucky Summer Fellowship for undergraduates (RB) and in part by NSF grants IBN-9808631 (RLC) and NSF-ILI-DUE 9850907 (RLC). Illustrations were provided by the courtesy of Ms Hye Won Cooper. Appreciation is given to Dr Grace Jones (Univ. of KY) for initial help in the use of a culture hood.

Keywords

  • Behavior
  • Development
  • Neuromuscular junction
  • Neurotransmission

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology

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