Studying properties of neurotransmitter receptors by non-stationary noise analysis of spontaneous postsynaptic currents and agonist-evoked responses in outside-out patches.
Chemical synaptic transmission depends on neurotransmitter-gated ion channels concentrated in the postsynaptic membrane of specialized synaptic contacts. The functional characteristics of these neurotransmitter receptor channels are important for determining the properties of synaptic transmission. Whole-cell recording of postsynaptic currents (PSCs) and outside-out patch recording of transmitter-evoked currents are important tools for estimating the single-channel conductance and the number of receptors contributing to the PSC activated by a single transmitter quantum. When single-channel activity cannot be directly resolved, non-stationary noise analysis is a valuable tool for determining these parameters. Peak-scaled non-stationary noise analysis can be used to compensate for quantal variability in synaptic currents. Here, we present detailed protocols for conventional and peak-scaled non-stationary noise analysis of spontaneous PSCs and responses in outside-out patches. In addition, we include examples of computer code for individual functions used in the different stages of non-stationary noise analysis. These analysis procedures require 3-8 h.
Hartveit
Espen
E
Department of Biomedicine, University of Bergen, Jonas Lies vei 91, N-5009 Bergen, Norway. espen.hartveit@biomed.uib.no
Veruki
Margaret Lin
ML
D000818:Animals; D015640:Ion Channel Gating / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D008959:Models, Neurological; D009482:Neurophysiology / Q000379:methods*; D018408:Patch-Clamp Techniques / Q000379:methods; D017981:Receptors, Neurotransmitter / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology*
D017981:Receptors, Neurotransmitter