title: Locust primary neuronal culture for the study of synaptic transmission.
abstract: We have designed a cell culture system for thoracic neurons of adult Locusta migratoria that enables the establishment of functional synapses in vitro. Patch-clamp recordings revealed three different neuron classes. About half of the neurons (47%) had unexcitable somata with outward and no inward conductance. The other half generated either single (37%) or multiple action potentials (18%) and differed mainly in lower outward conductance. Selectively stained motor neurons were analyzed to demonstrate varied physiological properties due to culture conditions. Using paired patch clamp recordings we demonstrate directly synaptic transmission in morphologically connected neurons in vitro. Presynaptic stimulation resulted in postsynaptic potentials in 42 pairs of neurons tested, independent of the type of neuron. According to pharmacological experiments most of these synapses were either glutamatergic or GABAergic. In addition to these chemical synapses, electrical synapses were found. With the demonstration of synapse formation in cell culture of adult locust neurons, this study provides the basis for the future analysis of more defined insect neuronal circuits in culture.
authors.affiliation: Peter Grünberg Institute, Institute of Complex Systems, Bioelectronics, Forschungszentrum Jülich, 52425, Jülich, Germany. stefan.weigel@wzw.tum.de
authors.forename: Stefan
authors.initials: S
authors.lastname: Weigel
authors.forename: Petra
authors.initials: P
authors.lastname: Schulte
authors.forename: Simone
authors.initials: S
authors.lastname: Meffert
authors.forename: Peter
authors.initials: P
authors.lastname: Bräunig
authors.forename: Andreas
authors.initials: A
authors.lastname: Offenhäusser
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000818:Animals; D018929:Cell Culture Techniques; D054351:Electrical Synapses / Q000187:drug effects / Q000502:physiology*; D005260:Female; D018682:GABA Agents / Q000494:pharmacology; D005724:Ganglia / Q000166:cytology / Q000502:physiology; D006110:Grasshoppers / Q000502:physiology*; D008297:Male; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D013268:Stimulation, Chemical; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*
chemical_list: D018682:GABA Agents