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The Journal of Neuroscience, August 31, 2005, 25(35):8104-8111; doi:10.1523/JNEUROSCI.2611-05.2005

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Cellular/Molecular
Paired Motor Neuron–Muscle Recordings in Zebrafish Test the Receptor Blockade Model for Shaping Synaptic Current

Hua Wen and Paul Brehm

Department of Neurobiology and Behavior, State University of New York at Stony Brook, Stony Brook, New York 11794

The transparent spinal cord and electrically compact fast muscle of zebrafish offer the first opportunity to perform simultaneous whole-cell patch-clamp recordings from both motor neuron and target skeletal muscle in situ. Our paired recordings reveal the fastest reported kinetics for both spontaneous and evoked synaptic currents at any synapse and a large quantal size that facilitates the resolution of spontaneous synaptic currents. We used this preparation to test the recent proposal that open channel block of the acetylcholine receptor by acetylcholine modulates the kinetics and timing of transmission between nerve and muscle in larval zebrafish (Legendre et al., 2000). Contrary to the predictions of this model, we find similar delay and onset kinetics of synaptic current at positive and negative muscle membrane potentials, even after inhibition of acetylcholinesterase. In contrast, blockade of motor neuron K channels by 4-aminopyridine prolonged the action potential and introduced a significant delay and slowing of evoked synaptic currents, demonstrating our ability to measured altered transmitter release with this system. We conclude that the kinetics of neuromuscular synaptic currents in zebrafish is not governed by receptor block.

Key words: neuromuscular; channel block; cholinergic; quantal analysis; patch clamp; spinal cord; K channel

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Received June 24, 2005; revised July 26, 2005; accepted July 26, 2005.

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