一种光遗传工具可提高单细胞的细胞内pH值并驱动局部膜动力学
Caitlin E T Donahue1,2, Michael D Siroky1,2, Katharine A White1,2
1Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, United States.
Journal of the American Chemical Society
|November 2, 2021
概括
研究人员使用Archaerhodopsin (ArchT) 开发了一种新工具,用于精确控制单细胞中的细胞内pH值 (pHi). 这种方法表明,高的pHi可以直接触发膜和其他动态细胞行为.
科学领域:
- 细胞生物学
- 生物物理
- 生理学
背景情况:
- 细胞内pH (pHi) 动态调节关键的细胞功能,包括偏离,细胞骨架重塑和迁移.
- 之前对pH依赖细胞行为的研究受限于种群级分析和非特异的pHi操纵方法.
- 缺乏单细胞分辨器工具阻碍了对pHi在单细胞行为中的作用的理解.
研究的目的:
- 开发和验证一种用于精确单细胞操纵细胞内pH值 (pHi) 的新工具.
- 调查pHi的升高是否足以在单个细胞中诱导膜.
- 探索pHi动态与单细胞行为之间的时空关系.
主要方法:
- 使用Archaerhodopsin (ArchT),一种光驱向外的质子,用于单细胞中的向pHi增加.
- 已证明ArchT可重复激活,使pHi持续升高 (长达45分钟).
- 应用ArchT工具来观察和量化对受控的pHi变化的膜和动态.
主要成果:
- 在很短的时间内,ArchT有效地诱导了强大的生理性PHY增加.
- 单细胞激活ArchT可以在几秒钟内迅速导致局部膜.
- 与对照组相比,提升的pHi显著增加了膜动态,包括突起和收缩事件.
结论:
- 考古罗多普辛 (ArchT) 是一个强大的空间时间控制单细胞pHi的工具.
- 细胞内pH值升高是启动膜和改变细胞膜动态的足够驱动因素.
- 这种技术可以直接研究pHi在各种单细胞行为中的作用,进步细胞生理学研究.
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