In yeast, adaptation to varying conditions often requires proper regulation of the plasma membrane potential. To determine yeast membrane potential change, optical methods involving potentiometric dyes have been supplemental to the direct electrode-based method. However, the hydrophobic nature of the dyes and their slow distribution across the membrane still limits their utilization. Genetically encoded voltage indicator (GEVI) proteins employed in neuroscience offer a tantalizing alternative for monitoring yeast membrane potential change. In this work, several widely used GEVI proteins were assessed in Saccharomyces cerevisiae for their expression and function as a voltage reporter. Among them, only ArcLight and Accelerated Sensor of Action Potential (ASAP) proteins could be expressed and transported to the plasma membrane. While the voltage-sensing capability was demonstrated for both ArcLight and ASAP, ArcLight fluorescence was sensitive to the intracellular pH change concurrently with the voltage change. Therefore, we established that ASAP is the more suitable GEVI protein for reporting yeast membrane potential change. This voltage-sensing reporter for yeast based on ASAP offers a new effective strategy for real-time optical detection of yeast membrane potential change, which potentially facilitates many areas of yeast research including optimizing growth conditions for industrial use and investigating yeast ion transport system.

中文翻译：

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使用基因编码的电压指示蛋白实时检测酵母质膜电位的变化。

在酵母中，适应不同条件通常需要适当调节质膜电位。为了确定酵母膜电位变化，涉及电位染料的光学方法已经补充了基于直接电极的方法。然而，染料的疏水性及其在整个膜上的缓慢分布仍然限制了它们的利用。神经科学中采用的基因编码电压指示剂 (GEVI) 蛋白为监测酵母膜电位变化提供了一种诱人的替代方案。在这项工作中，在酿酒酵母中评估了几种广泛使用的 GEVI 蛋白因为它们作为电压报告器的表达和功能。其中，只有 ArcLight 和 Accelerated Sensor of Action Potential (ASAP) 蛋白可以表达并转运到质膜。虽然 ArcLight 和 ASAP 都证明了电压传感能力，但 ArcLight 荧光对与电压变化同时发生的细胞内 pH 变化敏感。因此，我们确定 ASAP 是更适合报告酵母膜电位变化的 GEVI 蛋白。这种基于 ASAP 的酵母电压传感报告器为酵母膜电位变化的实时光学检测提供了一种新的有效策略，这可能促进酵母研究的许多领域，包括优化工业用途的生长条件和研究酵母离子传输系统。