In vivo whole-cell recording with high success rate in anaesthetized and awake mammalian brains
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METHODOLOGY
Open Access
In vivo whole-cell recording with high success rate in anaesthetized and awake mammalian brains Yao Wang†, Yu-zhang Liu†, Shi-yi Wang and Zhiru Wang*
Abstract As a critical technique for dissection of synaptic and cellular mechanisms, whole-cell patch-clamp recording has become feasible for in vivo preparations including both anaesthetized and awake mammalian brains. However, compared with in vitro whole-cell recording, in vivo whole-cell recording often suffers from low success rates and high access resistance, preventing its wide application in physiological analysis of neural circuits. Here, we describe experimental procedures for achieving in vivo amphotericin B-perforated whole-cell recording as well as conventional (breakthrough) whole-cell recording from rats and mice. The success rate of perforated whole-cell recordings was 70―80 % in the hippocampus and neocortex, and access resistance was 40―70 MΩ. The success rate of conventional whole-cell recordings was ~50 % in the hippocampus, with access resistance of 20―40 MΩ. Recordings were stable, and in awake, head-fixed animals, ~50 % whole-cell patched neurons could be held for > 1 hr. The conventional whole-cell recording also permitted infusion of pharmacological agents, such as intracellular blockers of Na+ channels and NMDA receptors. These findings open new possibilities for synaptic and cellular analysis in vivo. Keywords: In vivo, Whole-cell recording, Amphotericin, Anaesthetized, Awake, Cortex, Hippocampus
Introduction Whole-cell patch-clamp recording is an important neuroscience technique. It allows the measurement of both subthreshold electrical activity and suprathreshold firing of a cell. In addition, the membrane potential of the recorded cell can be altered by current injection for determining the existence and properties of specific ion channels or receptors, as well as for determining the effect of neuronal activity on neural circuit functions such as activityinduced synaptic plasticity. Moreover, whole-cell recording permits infusion of pharmacological agents in the cell to conduct intracellular pharmacological studies or label cell morphology. Due to the distinct advantages, the technique of whole-cell recording is very powerful for investigating the intrinsic and synaptic properties of a neuron. In addition to its early application for recordings from in * Correspondence: [email protected] † Equal contributors Institute and Key Laboratory of Brain Functional Genomics of Chinese Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics, School of Life Sciences, East China Normal University, Shanghai 200062, China
vitro preparations [1–4], whole-cell recording has been more recently applied to in vivo preparations [5]. Many groups have reported their findings with the use of in vivo whole-cell recording, as performed in anaesthetized animals [6–9] as well as in awake, head-fixed [7, 10, 11] or freely moving animals [9, 12], but the low success rate to achieve recordings and the high access
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