Abstract
The axon initial segment (AIS) is an essential neuronal compartment. It is usually where action potentials are initiated. Recent studies demonstrated that the AIS is a plastic structure that can be regulated by neuronal activity and by the activation of metabotropic receptors. Studying the AIS in live tissue can be difficult because its identification is not always reliable. Here we provide a new technique allowing a fast and reliable identification of the AIS in live brain slice preparations. By simultaneous recording of extracellular local field potentials and whole-cell patch-clamp recording of neurons, we can detect sinks caused by inward currents flowing across the membrane. We determine the location of the AIS by comparing the timing of these events with the action potential. We demonstrate that this method allows the unequivocal identification of the AIS of different types of neurons from the brain.
Highlights
The axon initial segment (AIS) is the gatekeeper of neurons
We have demonstrated a novel procedure allowing the online identification of the AIS during electrophysiological recording of neurons
Our method offers several advantages compared to others. It can be done in live tissue, while a neuron is recorded by means of the patch clamp technique. This facilitates the investigation of the physiological mechanisms involved in the modulation of the AIS
Summary
The axon initial segment (AIS) is the gatekeeper of neurons. In cartwheel neurons in the dorsal cochlear nucleus, the activation of dopamine D3 receptors at the AIS inhibits T-type calcium channels and thereby spike initiation (Bender et al, 2010). The activation of serotonergic 5-HT1A receptors at the AIS inhibits the Na+ current responsible for the genesis of action potentials. This mechanism is responsible for the central component of motor fatigue occurring during prolonged efforts (Cotel et al, 2013; Perrier and Cotel, 2015)
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