细菌脂多糖对甜味受体T1R2可变剪接与功能的调控作用

doi:10.7518/hxkq.2021.04.015

华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (4): 469-474.doi: 10.7518/hxkq.2021.04.015

细菌脂多糖对甜味受体T1R2可变剪接与功能的调控作用

朱健慧¹(), 郑欣¹, 彭显¹, 徐欣¹, Margolskee Robert², 周学东¹()

^1.口腔疾病研究国家重点实验室国家口腔疾病临床研究中心四川大学华西口腔医院牙体牙髓病科，成都 610041
^2.Monell Chemical Senses Center，美国费城 PA 19104

收稿日期:2020-07-18 修回日期:2021-04-19 出版日期:2021-08-01 发布日期:2021-08-10
通讯作者: 周学东 E-mail:240805997@qq.com;zhouxd@scu.edu.cn
作者简介:朱健慧，博士，E-mail：240805997@qq.com
基金资助:
国家自然科学基金(81900995);四川大学专职博士后研发基金(2019SCU12021);四川大学华西口腔医院人才队伍建设科研经费(RCDWJS2020-11);口腔疾病研究国家重点实验室开放课题(SKLOD202024)

Regulation effect of lipopolysaccharide on the alternative splicing and function of sweet taste receptor T1R2

Zhu Jianhui¹(), Zheng Xin¹, Peng Xian¹, Xu Xin¹, Margolskee Robert², Zhou Xuedong¹()

^1.State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Cariology and Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
^2.Monell Chemical Senses Center, Philadelphia PA 19104, USA

Received:2020-07-18 Revised:2021-04-19 Online:2021-08-01 Published:2021-08-10
Contact: Zhou Xuedong E-mail:240805997@qq.com;zhouxd@scu.edu.cn
Supported by:
The National Natural Science Foundation of China(81900995);The Postdoctoral Foundation of Sichuan University Grant(2019SCU12021);The Research Funding for Talents Developing, West China Hospital of Stomatology, Sichuan University(RCDWJS2020-11);The Open Fund of State Key Laboratory of Oral Diseases(SKLOD202024)

摘要/Abstract

摘要： 目的

寻找小鼠甜味受体T1R2的剪接异构体，探究细菌毒力因子脂多糖（LPS）对T1R2可变剪接及甜味受体功能的影响。

方法

分离提取小鼠味蕾组织RNA，逆转录聚合酶链式反应（RT-PCR）寻找小鼠甜味受体T1R2剪接异构体，通过体外异源性表达实验检测T1R2异构体对甜味受体功能的影响，通过实时荧光定量聚合酶链式反应检测LPS局部注射对T1R2异构体表达的影响。

结果

小鼠甜味受体T1R2存在剪接异构体T1R2_Δe3p，其与T1R3组成的甜味受体无法被甜味刺激物激活，且可显著下调本构体T1R2/T1R3的功能；LPS局部注射可使小鼠味蕾内T1R2_Δe3p表达比例明显上升。

结论

LPS刺激可影响小鼠甜味受体T1R2的可变剪接过程，显著上调无功能异构体T1R2_Δe3p的表达，提示T1R2可变剪接调控可能为微生物感染影响宿主味觉感知的机制之一。

关键词: 甜味受体, 可变剪接, 味觉感知, 脂多糖

Abstract: Objective

To identify the alternative splicing isoform of mouse sweet taste receptor T1R2, and investigate the effect of lipopolysaccharide (LPS) local injection on T1R2 alternative splicing and the function of sweet taste receptor as one of the bacterial virulence factors.

Methods

After mouse taste bud tissue isolation was conducted, RNA extraction and reverse transcription polymerase chain reaction (PCR) were performed to identify the splicing isoform of T1R2. Heterologous expression experiments in vitro were utilized to detect how the T1R2 isoform regulated the function of sweet taste receptors. The effect of local LPS injection on the expression of the T1R2 isoform was measured by real-time fluorescent quantitative PCR.

Results

T1R2 splicing isoform T1R2_Δe3p formed sweet taste receptors with T1R3, which could not be activated by sweet taste stimuli and significantly downregulated the function of canonical T1R2/T1R3. Local LPS injection significantly increased the expression ratio of T1R2_Δe3p in mouse taste buds.

Conclusion

LPS stimulation affects the alternative splicing of mouse sweet taste receptor T1R2 and significantly upregulates the expression of non-functional isoform T1R2_Δe3p, suggesting that T1R2 alternative splicing regulation may be one of the mechanisms by which microbial infection affects host taste perception.

Key words: sweet taste receptor, alternative splicing, taste perception, lipopolysaccharide

中图分类号:

Q 78

朱健慧, 郑欣, 彭显, 徐欣, Margolskee Robert, 周学东. 细菌脂多糖对甜味受体T1R2可变剪接与功能的调控作用[J]. 华西口腔医学杂志, 2021, 39(4): 469-474.

Zhu Jianhui, Zheng Xin, Peng Xian, Xu Xin, Margolskee Robert, Zhou Xuedong. Regulation effect of lipopolysaccharide on the alternative splicing and function of sweet taste receptor T1R2[J]. West China Journal of Stomatology, 2021, 39(4): 469-474.

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细菌脂多糖对甜味受体T1R2可变剪接与功能的调控作用

Regulation effect of lipopolysaccharide on the alternative splicing and function of sweet taste receptor T1R2

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