DREADDs-Based Chemogenetics Induced Slow Transit Constipation via Inhibition of Enteric Neurons

IF 2.3 3区医学 Q3 GASTROENTEROLOGY & HEPATOLOGY

Journal of Digestive Diseases Pub Date : 2025-04-14 DOI:10.1111/1751-2980.13344

Xin Yi Lu, Yu Xiang Wen, Ni Jiang, Si Qi Zhou, Tian Yang, Liang Liang Shi, Hui Min Guo, Wei Zhang, Qi Peng Zhang, Ni Na Zhang

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引用次数: 0

Abstract

Objectives

Designer receptors exclusively activated by designer drugs (DREADDs)-based chemogenetic tools are commonly used to activate or silence targeted neurons by the agonistic ligand deschloroclozapine (DCZ). This study aimed to establish a Gi-DREADD-based murine model of slow transit constipation (STC) and elucidate its pathophysiological mechanisms.

Methods

Adeno-associated virus (AAV) 9-hM4Di was injected into the intestinal wall of mice, and colonic motility was evaluated. The efficiency and immunogenicity of AAV9-hM4Di transduction in the enteric nervous system (ENS) were evaluated. Nitric oxide (NO), acetylcholine (ACh), and substance P (SP) in the colonic tissues and serum samples were analyzed. Calcium (Ca²⁺) imaging was performed to evaluate the responses of AAV9-hM4Di on enteric nerves.

Results

AAV9-hM4Di-treated mice showed gastrointestinal motility dysfunction, including reduced fecal pellets and decreased fecal mass and water content. Electrophysiological recording of muscle contraction in the isolated colonic tissues from the chemogenetic mice showed decreased frequency and amplitude after DCZ treatment. The mice treated with AAV9-hM4Di showed the highest levels of transduction in the myenteric plexuses of the ENS. There were no differences in transduction in neuronal nitric oxide synthase (nNOS) and choline acetyltransferase (ChAT) neurons. Gi-DREADDs significantly downregulated ACh but not NO or SP expression in the distal colon in the chemogenetic mice. Ca²⁺ transient in neurons of ENS in chemogenetic mice was strongly inhibited by DCZ.

Conclusions

It is feasible to apply the DREADDs-based chemogenetic tools to the ENS. Gi-DREADDs can selectively modulate the ENS, inducing STC without excitatory-neural bias, offering targeted neuromodulation for gastrointestinal motility disorders.

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基于dreadds的化学遗传学通过抑制肠道神经元诱导慢转运便秘

基于设计药物（DREADDs）的化学发生工具专门激活的设计受体通常用于通过激动性配体去氯氯氮平（DCZ）激活或沉默目标神经元。本研究旨在建立基于gi - dreadd的小鼠慢传输型便秘（STC）模型，并阐明其病理生理机制。方法将腺相关病毒（AAV） 9-hM4Di注入小鼠肠壁，观察其结肠运动。评价AAV9-hM4Di在肠神经系统（ENS）中的转导效率和免疫原性。测定大鼠结肠组织及血清中一氧化氮（NO）、乙酰胆碱（ACh）、P物质（SP）含量。钙（Ca2+）成像评价AAV9-hM4Di对肠神经的反应。结果aav9 - hm4di处理小鼠出现胃肠运动功能障碍，包括粪便颗粒减少，粪便质量和含水量降低。电生理记录显示，DCZ处理后小鼠离体结肠组织肌肉收缩的频率和幅度均下降。经AAV9-hM4Di处理的小鼠脑神经系统肌肠丛的转导水平最高，而神经元一氧化氮合酶（nNOS）和胆碱乙酰转移酶（ChAT）神经元的转导水平无差异。Gi-DREADDs显著下调化学发生小鼠远端结肠中ACh的表达，但不下调NO或SP的表达。DCZ强烈抑制趋化小鼠ENS神经元内Ca2+瞬态变化。结论基于dreadds的化学发生工具应用于ENS是可行的，Gi-DREADDs可以选择性地调节ENS，诱导STC，无兴奋性神经偏向，为胃肠道运动障碍提供靶向神经调节。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Digestive Diseases 医学-胃肠肝病学

CiteScore

5.40

自引率

2.90%

发文量

审稿时长

6-12 weeks

期刊介绍： The Journal of Digestive Diseases is the official English-language journal of the Chinese Society of Gastroenterology. The journal is published twelve times per year and includes peer-reviewed original papers, review articles and commentaries concerned with research relating to the esophagus, stomach, small intestine, colon, liver, biliary tract and pancreas.