Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Elsevier Science full text link Elsevier Science Free PMC article
Full text links

Actions

Share

.2021 Dec 1;2(4):100981.
doi: 10.1016/j.xpro.2021.100981. eCollection 2021 Dec 17.

Protocol for recording the discharge of locus coeruleus neurons in free-moving mice during different sleep-wake stages

Affiliations

Protocol for recording the discharge of locus coeruleus neurons in free-moving mice during different sleep-wake stages

Yue Liang et al. STAR Protoc..

Abstract

Cortical electroencephalography (EEG) is generally used to detect the different sleep-wake states of animals. EEG combined within vivo multichannel recording provides a powerful tool for decoding the neural network of sleep-wake regulation. Here, we detail a protocol using cortical EEG combined within vivo multichannel recording to examine the activity of locus coeruleus (LC) neurons in free-moving mice at different sleep-wake stages. The procedures for electrode fabrication, the surgery to implant electrodes, and post-recording data analysis are also included. For complete details on the use and execution of this protocol, please refer to Liang et al. (2021).

Keywords: Behavior; Cell Biology; Model Organisms; Neuroscience.

© 2021 The Authors.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Homemade multichannel electrodes (A) Accessories for making electrodes. (B) A 20-pin connector was soldered to a 20-pin circuit board. (C) Arranged insulated nichrome wires, two insulated EEG wires, two insulated EMG wires and two ground wires were soldered to the 20-pin circuit board. (D) The welds were sealed with AB glue. (E) The nichrome wires were cut to the appropriate length, and the anchoring part (the yellow dotted line in the picture) of the circuit board was removed. (F) A schematic diagram of the arrangement and distribution of electrode wires. Thirteen individually insulated nichrome wire arrays were arranged in a 3 + 3 + 3 + 3 + 1 pattern (approximately 200 μm between lines), one of which was a separate arrangement of the reference electrode.
Figure 2
Figure 2
The surgical procedure for implanting electrodes (A) Adult mice were anaesthetized, the head was fixed in a stereotaxic frame, and the hair on the head of the mouse was removed. (B) Two skull screws were inserted in the frontal skull for EEG recording, and two others were placed in the skull of the lateral parietal region for grounding. The skull was leveled using a stereotaxic frame, and the location of the target nucleus was marked. Scaling bar: 5 mm. (C) The electrode bundle of the homemade multichannel electrode was slowly inserted into a hole at the targeted location on the skull. (D) The electrode bundle was secured by dental cement. (E) Two EEG wires were soldered to two skull screws in the frontal region for EEG recording, and two ground wires were soldered to two other screws for grounding. (F) A large spherical solder was soldered at the terminals of the EMG wires. (G) Two EMG wires were inserted into the neck muscles, and the skin was sutured. (H) Dental cement was used to secure the exposed skull and electrode. (I) The mouse was recovered after surgery.
Figure 3
Figure 3
EEG-EMG signal recording and LC neuronal spike acquisition (A) Recording system (Zeus, Bio-Signal Technologies, McKinney, TX, USA). (B) The freely moving mouse in the shielding box with the head connected to the recording system. (C) The connection of the recording apparatus included the headstage, adaptor and electrode. (D) (Left) LFP and (right) spike signals were recorded.
Figure 4
Figure 4
LFP and spike signals data processing (A) Definition of LFP signal as wakefulness, NREM sleep, or REM sleep in 4-s epochs using SleepSign. (B) EEG and EMG recording traces during wakefulness, NREM sleep, and REM sleep. (C) Action potential waveforms of two sorted LC neurons from a single electrode channel and clusters formed by the distinct action potential waveforms plotted in principal component space are shown in Offline Sorter.
Figure 5
Figure 5
Action potential waveforms and clusters of LC unit spikes in different stages of sorting (A) Action potential waveforms of LC unit in the initial file. (B) Action potential waveforms of LC unit after remove the obviously distinct, mussy waves manually. (C) Action potential waveforms of LC unit after the completion of sorting. (D) Clusters formed by the distinct action potential waveforms plotted in different principal component spaces in the initial file. (E) Clusters formed by the distinct action potential waveforms plotted in different principal component spaces after the completion of sorting.
Figure 6
Figure 6
Evaluation of recording quality and success probability of LC discharge recording by multichannel electrode Left: Statistics of the number of various channels and LC units per mouse (n = 4 mice). Right: The proportion of successful channels and failed channels in all available channels of 4 mice.
Figure 7
Figure 7
The amplitude and waveform width of LC unit spike Left: The mean (± S.D.) amplitude of LC unit spike was 0.1938 ± 0.1848 mV (cells = 29 from 4 mice). Right: The mean (± S.D.) waveform width of LC unit spike was 0.1322 ± 0.04858 ms (cells = 29 from 4 mice).
Figure 8
Figure 8
LC neuronal activity during different sleep stages (A) A brain slice from a mouse with electrodes implanted in the LC. Blue arrow indicates the electrode track. Scaling bar: 500 μm. (B) Representative EEG power spectrum, EMG trace and firing rates of an example LC neuron during different brain states (color coded). (C) Average firing rate of 29 LC neurons in each state. Each line shows firing rates of one unit. (∗∗∗p < 0.001, cells = 29 from 4 mice). Data are represented as the mean ± SEM. The statistical significance of the data was calculated with one-way ANOVA.
Figure 9
Figure 9
Distribution of multiple unit signals in different 2D cluster views Left: Action potential waveforms of three sorted LC neurons from a single electrode channel. Right: Clusters formed by the distinct action potential waveforms plotted in different principal component spaces.
Figure 10
Figure 10
Interspike interval (ISI) histograms of LC neuron Top: ISI histograms of LC neurons with successful noise removal. Bottom: ISI histograms of LC neurons with incomplete noise removal.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Fukunaga K., Mantock J.M. Nonparametric discriminant analysis. IEEE Trans. Pattern Anal. Mach. Intell. 1983;5:671–678. - PubMed
    1. Lewicki M.S. A review of methods for spike sorting: the detection and classification of neural action potentials. Network. 1998;9:R53–R78. - PubMed
    1. Li Z.H., Yang Y., Hou M.M., Wang Y., Qin X., Zhang C., Zhang Y. The application of in vivo multi-channel recording methods in the studies of the affective pain in rats. Chin. J. Appl. Physiol. 2016;32:41–45. - PubMed
    1. Liang Y., Shi W., Xiang A., Hu D., Wang L., Zhang L. The NAergic locus coeruleus-ventrolateral preoptic area neural circuit mediates rapid arousal from sleep. Curr. Biol. 2021;31:3729–3742.e5. - PubMed
    1. Luo Y.J., Li Y.D., Wang L., Yang S.R., Yuan X.S., Wang J., Cherasse Y., Lazarus M., Chen J.F., Qu W.M., et al. Nucleus accumbens controls wakefulness by a subpopulation of neurons expressing dopamine D(1) receptors. Nat. Commun. 2018;9:1576. - PMC - PubMed

Publication types

MeSH terms

LinkOut - more resources

Full text links
Elsevier Science full text link Elsevier Science Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp