Journal
JOURNAL OF NEUROSCIENCE METHODS
Volume 183, Issue 2, Pages 158-164Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2009.06.025
Keywords
Action potential; Physiology; Neuronal excitability; Current injection; Current clamp; Synapse; MNTB; Calyx of Held
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Funding
- MRC
- MRC [MC_U132681855] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BBS/B/07276] Funding Source: researchfish
- Medical Research Council [MC_U132681855] Funding Source: researchfish
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Most current clamp studies trigger action potentials (APs) by step current injection through the recording electrode and assume that the resulting APs are essentially identical to those triggered by orthodromic synaptic inputs. However this assumption is not always valid, particularly when the synaptic conductance is of large magnitude and of close proximity to the axon initial segment. We addressed this question of similarity using the Calyx of Held/MNTB synapse; we compared APs evoked by long duration step current injections, short step current injections and orthodromic synaptic stimuli. Neither injected current protocol evoked APs that matched the evoked orthodromic AP waveform, showing differences in AP height, half-width and after-hyperpolarization. We postulated that this 'error' could arise from changes in the instantaneous conductance during the combined synaptic and AP waveforms, since the driving forces for the respective ionic currents are integrating and continually evolving over this time-course. We demonstrate that a simple Ohm's law manipulation of the EPSC waveform, which accounts for the evolving driving force on the synaptic conductance during the AP, produces waveforms that closely mimic those generated by physiological synaptic stimulation. This stimulation paradigm allows supra-threshold physiological stimulation (single stimuli or trains) without the variability caused by quantal fluctuation in transmitter release, and can be implemented without a specialised dynamic clamp system. Combined with pharmacological tools this method provides a reliable means to assess the physiological roles of postsynaptic ion channels without confounding affects from the presynaptic input. (C) 2009 Published by Elsevier B.V.
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