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The Journal of Neuroscience, July 1, 1999, 19(13):5311-5321
Fast Excitatory Synaptic Transmission Mediated by Nicotinic
Acetylcholine Receptors in Drosophila Neurons
Daewoo
Lee and
Diane K.
O'Dowd
Departments of Developmental and Cell Biology, Anatomy and
Neurobiology, University of California Irvine, Irvine, California
92697-1280
Difficulty in recording from single neurons in vivo
has precluded functional analyses of transmission at central synapses in Drosophila, where the neurotransmitters and receptors
mediating fast synaptic transmission have yet to be identified. Here we demonstrate that spontaneously active synaptic connections form between
cultured neurons prepared from wild-type embryos and provide the first
direct evidence that both acetylcholine and GABA mediate fast
interneuronal synaptic transmission in Drosophila. The
predominant type of fast excitatory transmission between cultured
neurons is mediated by nicotinic acetylcholine receptors (nAChRs).
Detailed analysis of cholinergic transmission reveals that spontaneous EPSCs (sEPSCs) are composed of both evoked and action
potential-independent [miniature EPSC (mEPSC)] components. The mEPSCs
are characterized by a broad, positively skewed amplitude histogram in
which the variance is likely to reflect differences in the currents
induced by single quanta. Biophysical characteristics of the
cholinergic mEPSCs include a rapid rise time (0.6 msec) and decay ( = 2 msec). Regulation of mEPSC frequency by external calcium and cobalt
suggests that calcium influx through voltage-gated channels influences the probability of ACh release. In addition, brief depolarization of
the cultures with KCl can induce a calcium-dependent increase in sEPSC
frequency that persists for up to 3 hr after termination of the
stimulus, illustrating one form of plasticity at these cholinergic
synapses. These data demonstrate that cultured embryonic neurons,
amenable to both genetic and biochemical manipulations, present a
unique opportunity to define genes/signal transduction cascades
involved in functional regulation of fast excitatory transmission at
interneuronal cholinergic synapses in Drosophila.
Key words:
Drosophila; nAChRs; interneuronal synapses; mEPSC; EPSC; fast excitatory cholinergic transmission
Copyright © 1999 Society for Neuroscience 0270-6474/99/19135311-11$05.00/0
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