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Neuroscience
末梢神経刺激研究のためのin vivo胸部背根神経節(DRG)カルシウムイメージングとECG記録
末梢神経刺激研究のためのin vivo胸部背根神経節(DRG)カルシウムイメージングとECG記録
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
In Vivo Thoracic Dorsal Root Ganglia (DRG) Calcium Imaging and ECG Recording for Studying Peripheral Nerve Stimulation

末梢神経刺激研究のためのin vivo胸部背根神経節(DRG)カルシウムイメージングとECG記録

Full Text
1,899 Views
06:34 min
August 16, 2024

DOI: 10.3791/67283-v

Xia Li1, Yun Liu1, Kun Liu1, Longhua Du1, Tao Lv1, Bing Zhu1, Xinyan Gao1

1Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a novel surgical manipulation to expose the thoracic dorsal root ganglia (DRG) in anesthetized mice, facilitating in vivo calcium imaging alongside synchronized electrocardiogram (ECG) recording. The research investigates whether acupuncture at the PC6 point can activate DRG neurons while also regulating ECG signals, providing valuable insights into the interactions of peripheral nervous system and visceral organ inputs.

Key Study Components

Area of Science

  • Neuroscience
  • Electrophysiology
  • Biophysics

Background

  • Thoracic DRGs are challenging to expose due to anatomical and physiological factors.
  • Current acupuncture studies utilize various methods, including electrophysiology and neural tracing.
  • The use of in vivo calcium imaging enhances understanding of DRG neuron activity.
  • The study aims to address the gap in knowledge regarding the effects of acupuncture on DRG neurons.

Purpose of Study

  • To explore the activation of DRG neurons through acupuncture at specific points.
  • To assess the impact of acupuncture on cardiac rhythms via ECG monitoring.
  • To provide a methodological framework for future studies on neuronal interactions and modalities.

Methods Used

  • In vivo calcium imaging of thoracic DRGs was employed.
  • The biological model consists of anesthetized mice with surgical exposure of thoracic dorsal root ganglia.
  • Specific protocols included securing the mice, exposing the DRG, and electrophysiological assessments.
  • Control measures involved maintaining the mice under respiratory anesthesia during experimentation.
  • Real-time imaging and stimulation techniques were utilized to assess neuronal responses.

Main Results

  • Acupuncture at the PC6 point activated DRG neurons, evidenced by increased GCaMP fluorescence.
  • Both somatic and acupuncture stimulations yielded similar neuronal responses in terms of fluorescence activity.
  • Heart rate changes were correlated with DRG neuron activity during peripheral nerve stimulations.
  • This study marked the first in vivo observation of thoracic DRG neuron activities.

Conclusions

  • This research establishes a reliable method for observing in vivo DRG activities in response to acupuncture.
  • The implications of the findings enhance our understanding of how peripheral stimulations can influence both sensory and cardiac functions.
  • Future studies will explore the role of various receptors in acupuncture efficacy and the crosstalk between somatic and visceral neuronal systems.

Frequently Asked Questions

What are the advantages of the in vivo calcium imaging method?
In vivo calcium imaging allows for real-time monitoring of neuronal activities, providing dynamic insights into how neurons respond to different stimuli.
How is the thoracic DRG exposure achieved during the procedure?
The exposure involves a surgical incision and careful removal of muscle layers, ensuring minimal disruption to surrounding structures while preserving the integrity of the DRG.
What type of data is collected during the experiments?
Data include neuronal fluorescence responses, ECG readings, and heart rate fluctuations associated with different stimulation protocols.
Can this method be adapted for other types of neuromodulation studies?
Yes, the surgical approach and imaging techniques can be adapted for various interventions, allowing exploration of other neuromodulatory effects.
What limitations should be considered when interpreting the results?
Limitations include the physiological complexity of the thoracic region and potential variations in individual animal responses to acupuncture stimulation.

この研究では、麻酔をかけたマウスでT-DRGを曝露するための外科的操作を示し、 in vivo カルシウムイメージングと同期ECG記録を行います。この方法は、末梢電気神経刺激と胸部内臓入力、およびそれらの一次感覚レベルでの相互作用を研究するための最先端のツールです。

この研究では、麻酔をかけたマウスの胸部DRGを露出させる外科的操作を導入し、同期心電図記録とともにin vivoカルシウムイメージングを行いました。これにより、孔のPC6での鍼治療がDRGニューロンを活性化し、同時に心電図を調節できるかどうかに答えようとします。現在、鍼治療研究で使用されている技術には、in vivoまたはvitroの電気生理学、神経追跡、および光遺伝学と化学遺伝学を組み合わせたさまざまな戦略が含まれます。

この研究で紹介されたin vivoカルシウムイメージングは、鍼治療によって誘発されるDRGニューロン集団の活動を、すべて異なる動物モデルで動的かつリアルタイムに記録することにより、より深く理解するのに役立ちます。胸椎の生理学的湾曲により、胸椎の1〜5つの椎体セグメントで、胸部DRGを露出させることは非常に困難です。さらに、心拍リズムと肺リズムの影響により、胸部DRGの固定が困難になります。

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生体内カルシウムイメージング 胸部背根神経節 DRG 末梢神経刺激 体細胞刺激 神経活動 自律神経機能 内臓痛 体性心反射 心電図 T1-DRG 心機能の同期

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