JoVE Encyclopedia of Experiments
Neuroscience
0 views • 5:18 min • June 17th, 2025
Take a paralyzed transgenic zebrafish larva mounted in a perfusion chamber.
The larva expresses membrane-localized calcium indicators in the hair cells of the neuromast, a component of the lateral-line system that detects water movement.
The hair cells comprise apical hair bundles and basal presynaptic regions, which form synapses with afferent neurons to transmit signals.
Under a confocal microscope, use low magnification to position a fluid-jet pipette near the larva..
Increase magnification to locate a neuromast, reposition the pipette near it, and switch to
fluorescence imaging.
Deliver fluid pulses to deflect hair bundles, opening mechanotransduction channels for apical cation influx, including calcium ions.
Weakly fluorescent indicators bind to calcium, causing a localized increase in fluorescence.
The cation influx depolarizes the cell, triggering calcium entry through voltage-gated channels at the basal presynapse, where it binds to the indicators and increases fluorescence locally.
Visualize the apical and basal calcium dynamics in real time.
In this procedure, backfill 10 microliter of neuronal buffer into a properly broken fluid jet needle using a gel loading tip. Load the soluti
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