In vivo whole-cell recording with high success rate in anaesthetized and awake mammalian brains.
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.
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.
Yao
Y
Wang
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.
Yu-Zhang
YZ
Liu
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.
Shi-Yi
SY
Wang
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. zrwang@brain.ecnu.edu.cn.
Zhiru
Z
Wang
Amphotericin; Anaesthetized; Awake; Cortex; Hippocampus; In vivo; Whole-cell recording
Pflugers Arch. 1981 Aug;391(2):85-100
6270629
Neuroreport. 1991 Aug;2(8):485-8
1912484
Pflugers Arch. 1989 Sep;414(5):600-12
2780225
Nature. 2009 Oct 15;461(7266):941-6
19829374
J Neurosci Methods. 1989 Dec;30(3):203-10
2607782
Nat Protoc. 2016 Apr;11(4):634-54
26938115
Nat Protoc. 2014 Dec;9(12):2784-95
25375992
J Physiol. 2011 Jul 1;589(Pt 13):3125-33
21540344
Neuron. 2014 Mar 19;81(6):1274-81
24560703
Nat Neurosci. 2014 Jun;17(6):841-50
24747575
J Physiol. 1990 Nov;430:605-16
1982315
Pflugers Arch. 2002 Jul;444(4):491-8
12136268
Front Neural Circuits. 2015 May 22;9:23
26052270
Neuron. 2006 Aug 17;51(4):399-407
16908406
J Neurosci Methods. 1991 Mar;37(1):15-26
2072734