AAV-hM3Dq and AAV-hM4Di were used for chemogenetics experiment. (From
BrainVTA)
The viruses used in this article from BrainVTA are in the table below
|
Chemogenetics |
PT-0019 AAV2/9-hSyn-DIO-hM3D(Gq)-mCherry
PT-0020 AAV2/9-hSyn-DIO-hM4D(Gi)-mCherry |
|
Control |
PT-0115 AAV2/9-hSyn-DIO-mCherry |
Yaochao Zheng, Shaoling Wu, Qian Yang, Zhen Xu, Subo Zhang, Shengnuo Fan, Cuicui Liu, Xiao Li, Chao Ma
Pub Date: 2021-01-13,
DOI: 10.1016/j.brainresbull.2021.01.008,
Email: [email protected]
Background: To determine if trigeminal nerve electrical stimulation (TNS) would be an effective arousal treatment for loss of consciousness (LOC), we applied neuroscientific methods to investigate the role of potential brain circuit and neuropeptide pathway in regulating level of consciousness.
Methods: Consciousness behavioral analysis, Electroencephalogram (EEG) recording, Chemogenetics, Microarray analysis, Milliplex MAP rat peptide assay, Chromatin immune-precipitation (ChIP), Dual-luciferase reporter experiment, Western blot, PCR and Fluorescence in situ hybridization (FISH).
Results: TNS can markedly activate the neuronal activities of the lateral hypothalamus (LH) and the spinal trigeminal nucleus (Sp5), as well as improve rat consciousness level and EEG activities. Then we proved that LH activation and upregulated neuropeptide hypocretin are beneficial for promotion of consciousness recovery. We then applied gene microarray experiment and found hypocretin might bemediated by a well-known transcription factor Early growth response gene 1 (EGR1), and the results were confirmed by ChIP and Dual-luciferase reporter experiment.
Conclusion: This study illustrates that TNS is an effective arousal strategy Treatment for LOC state via the activation of Sp5 and LH neurons and upregulation of hypocretin expression.
In the present study, the authors examined the effects of TNS on the behavioral levels and EEG characteristics of rats in TBI-induced LOC, through increasing the expression of transcription factor EGR1 and its down-stream target neuropeptide hypocretin. They then further explored and confirmed the potential brain regions which might participate in the therapeutic effects of TNS treatment.
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