An Optogenetics-based Approach to Regulate Colonic Contractions by Modulating the Activity of the Interstitial Cells of Cajal in Mice.

IF 3.3 3区 医学 Q2 CLINICAL NEUROLOGY
Song Zhao, Weidong Tong
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Abstract

Background/aims: The interstitial cells of Cajal (ICC) are pacemaker cells in the gastrointestinal (GI) tract. We examined whether the activity of ICC could be stimulated to control colonic contractions. An optogenetics-based mouse model in which the light-sensitive protein channelrhodopsin-2 (ChR2) was expressed was used to accomplish cell specific, direct stimulation of ICC.

Methods: An inducible site-specific Cre-loxP recombination system was used to generate KitCreERT2/+;ROSAChR2(H134R)/tdTomato/+ mice in which ChR2(H134R), a variant of ChR2, was genetically expressed in ICC after tamoxifen administration. Genotyping and immunofluorescence analysis were performed to confirm gene fusion and expression. Isometric force recordings were performed to measure changes in contractions in the colonic muscle strips.

Results: ChR2 was specifically expressed in Kit-labeled ICC. The isometric force recordings showed that the contractions of the colonic muscle strips changed under 470 nm blue light. Light stimulation evoked premature low-frequency and high amplitude (LFHA) contractions and enhanced the frequency of the LFHA contractions. The light-evoked contractions were blocked by T16Ainh-A01, an antagonist of anoctamin 1 channels that are expressed selectively in ICC in colonic muscles.

Conclusions: Our study demonstrates a potentially feasible approach to stimulate the activity of ICC by optogenetics. The colonic motor patterns of muscle strips, especially LFHA contractions, can be regulated by 470 nm light via ChR2, which is expressed in ICC.

Abstract Image

Abstract Image

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基于光遗传学的方法通过调节小鼠Cajal间质细胞的活性来调节结肠收缩。
背景/目的:Cajal间质细胞(ICC)是胃肠道中的起搏器细胞。我们研究了是否可以刺激ICC的活性来控制结肠收缩。利用光遗传学的小鼠模型,表达光敏蛋白通道视紫红质-2 (ChR2)来完成细胞特异性的直接刺激ICC。方法:采用诱导位点特异性Cre-loxP重组系统,生成KitCreERT2/+;ROSAChR2(H134R)/tdTomato/+小鼠,在给药后,在ICC中基因表达ChR2(H134R)。基因分型和免疫荧光分析证实基因融合和表达。进行等长力记录以测量结肠肌条收缩的变化。结果:ChR2在kit标记的ICC中特异性表达。等长力记录显示,在470 nm蓝光照射下,结肠肌条收缩发生变化。光刺激可诱发低频高振幅(LFHA)过早收缩,并增强LFHA收缩的频率。光诱发的收缩被T16Ainh-A01阻断,T16Ainh-A01是一种氨基酸1通道拮抗剂,在结肠肌肉的ICC中选择性表达。结论:我们的研究证明了光遗传学刺激ICC活性的潜在可行方法。肌条的结肠运动模式,特别是LFHA的收缩,可以通过在ICC中表达的ChR2在470 nm光下调节。
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来源期刊
Journal of Neurogastroenterology and Motility
Journal of Neurogastroenterology and Motility GASTROENTEROLOGY & HEPATOLOGY-CLINICAL NEUROLOGY
CiteScore
6.30
自引率
8.80%
发文量
96
期刊介绍: Journal of Neurogastroenterology and Motility (J Neurogastroenterol Motil) is a joint official journal of the Korean Society of Neurogastroenterology and Motility, the Thai Neurogastroenterology and Motility Society, the Japanese Society of Neurogastroenterology and Motility, the Indian Motility and Functional Disease Association, the Chinese Society of Gastrointestinal Motility, the South East Asia Gastro-Neuro Motility Association, the Taiwan Neurogastroenterology and Motility Society and the Asian Neurogastroenterology and Motility Association, launched in January 2010 after the title change from the Korean Journal of Neurogastroenterology and Motility, published from 1994 to 2009.
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