Gradient Tracking by Yeast GPCRs in a Microfluidics Chamber.

Cells typically exist in a highly dynamic environment, which cannot easily be recreated in culture dishes or microwell plates. Microfluidic devices can provide precise control of the time, dose, and orientation of a stimulus, while simultaneously capturing quantitative single-cell data. The approach is particularly powerful when combined with the genetically tractable yeast model organism. The GPCR pathway in yeast is structurally conserved and functionally interchangeable with those in humans. We describe the implementation of a microfluidic device to investigate morphological and transcriptional responses of yeast to a gradient or pulse administration of a GPCR ligand, the peptide mating pheromone α-factor.

细胞通常存在于一个高度动态的环境中，这种环境很难在培养皿或微孔板中轻易重现。微流控装置能够精确控制刺激的时间、剂量和方向，同时获取定量的单细胞数据。当与基因可操作的酵母模式生物相结合时，这种方法尤其强大。酵母中的G蛋白偶联受体（GPCR）通路在结构上是保守的，并且在功能上与人类的那些通路可互换。我们描述了一种微流控装置的应用，以研究酵母对一种GPCR配体——肽类交配信息素α因子的梯度或脉冲给药的形态和转录反应。