Drosophila NMJ electrophysiology analyses

Conventional intracellular recordings for assessing NMJ synaptic transmissions were carried out as previously described (55). Briefly, wandering third-instar larvae were dissected in calcium-free HL3.1 saline and recorded in HL3.1 saline containing 0.6 mM Ca2+ using intracellular microelectrodes (10 to 20 megohms) filled with 3 M KCl. Recordings were performed at 20° to 22°C with an Axoclamp 2B amplifier (Molecular Devices) in bridge mode, and the recorded data were processed with pCLAMP version 10.2 software (Molecular Devices). Both EJPs and mEJPs were recorded from muscle 6 of abdominal segment A3. EJPs were evoked by a Grass S48 stimulator (Astro-Grass Inc.) with suprathreshold stimulating pulses at 0.3 Hz, three EJP responses were collected for each animal, and mEJPs were recorded for a period of 60 s after the EJP recording. The data for analyses were recorded from cells with resting membrane potentials ranging from −60 to −65 mV.

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