Real-time Analysis of Gut-brain Neural Communication: Cortex wide Calcium Dynamics in Response to Intestinal Glucose Stimulation

Serika Yamada; Hiromu Monai

doi:10.3791/65902

JoVE Journal
Neuroscience

This content is Free Access.

JoVE Journal Neuroscience

Real-time Analysis of Gut-brain Neural Communication: Cortex wide Calcium Dynamics in Response to Intestinal Glucose Stimulation

肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学

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07:29 min

December 29, 2023

DOI: 10.3791/65902-v

Serika Yamada¹, Hiromu Monai¹

¹Department of Biology, Faculty of Science,Ochanomizu University

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Overview

This study investigates gut-brain communication through the vagus nerve, focusing on the effects of intragastric glucose injection on cortical activity in mice. The research highlights a novel method of catheter attachment to the gut, minimizing surgical trauma, facilitating the examination of neural communication mechanisms between the gut and brain.

Key Study Components

Area of Science

Neuroscience
Gastroenterology
Experimental Medicine

Background

Gut-brain communication is essential for processing information related to food preferences.
Traditionally, the vagus nerve has been treated as a singular entity, yet recent findings suggest it exhibits selective properties.
Understanding the mechanisms of stress on gut-brain signaling is crucial for exploring this communication pathway.

Purpose of Study

To observe the effects of intragastric glucose injections on cortical activity.
To develop a less invasive method for attaching catheters to the gut.
To explore how physical and psychological stress can impact gut-brain neural communication.

Methods Used

Mouse model used for the experiments, focusing on surgical techniques for catheter attachment.
Modification of traditional surgical methods by replacing sutures with cyanoacrylate glue for catheter attachment.
Imaging techniques employed to observe changes in cortical calcium dynamics following glucose administration.
Temporal fluorescence intensity changes were recorded post-injection for data analysis.

Main Results

Glucose injection into the duodenum led to significant changes in calcium dynamics in the secondary motor cortex.
Spontaneous calcium oscillations were recorded, illustrating patterns of burst suppression.
Imaging results indicated no significant changes following water administration, underscoring the specific effects of glucose.

Conclusions

The study provides insights into the rapid modulation of cortical activity by gut-derived signals.
Findings have implications for understanding the mechanisms underlying gut-brain interactions and their impact on neuronal activity.

Frequently Asked Questions

What advantages does the new catheter attachment method offer?

The new method using cyanoacrylate glue reduces surgical trauma compared to traditional suturing techniques, making it less invasive and more cost-effective.

How is the glucose injection administered in the study?

Intragastric glucose is injected through a catheter that is securely attached to the stomach, allowing for precise delivery and observation of cortical activity changes.

What types of data are obtained from the imaging techniques?

Data on cortical calcium dynamics are obtained, including spontaneous calcium oscillations and fluorescence intensity changes following glucose administration.

Could this methodology be adapted for other interventions?

Yes, the technique may be applicable to various pharmacological or biological interventions involving gut-brain signaling pathways.

What limitations should be considered with the surgical method?

While the method reduces trauma, careful handling and precise techniques are necessary to maintain the integrity of the gastrointestinal tract and surrounding tissues.

What implications does the study have for understanding gut-brain communication?

The findings enhance our comprehension of how gut-derived signals influence brain activity and may further explore the relationship between stress and gut-brain neural communication.

How does gut-brain communication relate to food preference?

Gut-brain communication plays a critical role in regulating food preferences and behaviors, highlighting the influence of the gastrointestinal system on neural processing related to diet.

由迷走神经促进的肠脑通讯对于胃肠道内分泌系统和大脑之间的通讯至关重要。然而，胃内葡萄糖注射是否可以改变皮质活性仍不清楚。在这里，我们提供了一个全面的方案来观察葡萄糖注射到十二指肠后皮质活动的变化。

我们的研究范围揭示了肠脑神经通讯，尤其是它在食物偏好中的作用。涉及迷走神经的实验通常被视为单个束，但最近的研究表明，它选择特性和器官特异性。所以我认为，调查它的统计数据，你知道的，将来会引起人们的注意。

这种将导管连接到肠道的方法比之前提出的方法适用，成本低，侵入性小，并且更容易。传统上，导管连接到肠道是通过缝合完成的。在这里，我们通过用氰基丙烯酸酯胶附件代替缝合来减轻对小鼠的手术损伤。

在我们的实验室，我们正在探索身体和心理压力如何影响小鼠肠脑神经通讯的机制。首先，用剪刀将硅管剪成 7 厘米的精确长度。使用氰基丙烯酸酯胶，固定距离硅管末端约 3 毫米的小塑料珠。

切除 23 号针的尖端，并从针尖切下 1.5 厘米。将针头的切割部分插入珠子另一侧的硅胶管中。使用钳子，从针尖切开 1 厘米。

将改良的 23 号注射针连接到 2.5 毫升注射器上。在 15 号注射针头上套上一根 23 厘米的硅管。从尖端切下注射针 1.5 厘米，并将其连接到硅管上。

首先，将麻醉的小鼠仰卧在手术台上，使其嘴巴靠近吸入装置。使用胶带将小鼠的口腔、前腿和后腿固定在手术台上。涂抹脱毛膏去除左上腹部的毛发。

在腹部右侧和剑突下方 5 毫米处做一个 1.5 厘米的皮肤切口。然后在腹壁上与初始皮肤切口相同的位置创建一个 1.5 厘米的切口。用钝头镊子轻轻横向移动左肝叶，露出胃。

现在，提起胃并通过切口轻轻将其取出。使用剪刀在幽门窦上创建一个小的穿孔。将带有珠子的导管末端引入穿孔。

确认导管牢固地连接到胃部后，小心地将胃重新定位到原来的位置。缝合腹壁，让导管从外部排出。然后以类似于腹部闭合的方式闭合皮肤切口。

用葡萄糖酸氯己定溶液清洁手术区域，并将小鼠放入消毒的笼子中。首先，使用辅助耳杆将麻醉的鼠标固定在立体定位平台上，以减轻脉动和呼吸的影响。使用电动剃须刀或脱毛霜，小心地去除头皮上的毛发。

用 0.1 至 0.5% 葡萄糖酸氯己定溶液消毒头皮表面。将局部麻醉凝胶涂抹在头皮上，等待 5 到 10 分钟。接下来，用剪刀从后脑勺到前额笔直剪出。

使用夹子拉回露出颅骨的多余皮肤。用棉签去除骨膜的结缔组织。立即将丙烯酸水泥涂抹在颅骨上，等待 5 分钟让水泥干燥。

在荧光立体显微镜下移动鼠标。对于成像，请将宽带蓝色荧光滤光片组与汞光源结合使用。接下来，从导管末端取下导管针头。

使用大约 0.03 毫升生理盐水清除小鼠侧导管内的任何残留内容物。然后从小鼠身上取下导管。将适当剂量的 10% 葡萄糖溶液吸入注射器中，并将其连接到导管上。

在成像软件中，检查相机识别并将帧速率设置为 10 赫兹。将分辨率设置为 512 x 512 像素，将深度设置为 16 位。单击记录过程按钮并获取 50 秒的自发数据。

最后，随着葡萄糖溶液的逐渐滴注，记录小鼠的生理状态数据。自发的神经活动显示整个皮层的随机钙振荡。时间荧光强度变化显示钙振荡遵循突发抑制模式。

葡萄糖注射在葡萄糖给药完成后 4 至 8 秒内显示皮质钙动力学发生显着变化，次级运动皮层立即激活。然而，在加水后没有观察到任何变化。与水给药相比，葡萄糖注射后在次级运动皮层区域观察到荧光强度比的显着变化。

注射后不同皮层区域的激活水平仅在次级运动皮层区域表现出显着差异。

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关键词:肠脑通讯葡萄糖刺激钙动力学全皮层成像实时分析胃内葡萄糖注射转基因小鼠基因编码的钙指标神经通路皮层活动