PDF(Pubmed)
Abstract：
Utilizing a microfluidic chip with serpentine channels, we inoculated the chip with an agar plug with Neurospora crassa mycelium and successfully captured individual hyphae in channels. For the first time, we report the presence of an autonomous clock in hyphae. Fluorescence of a mCherry reporter gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously along hyphae every half an hour for at least 6 days. We entrained single hyphae to light over a wide range of day lengths, including 6,12, 24, and 36 h days. Hyphae tracked in individual serpentine channels were highly synchronized (K = 0.60-0.78). Furthermore, hyphae also displayed temperature compensation properties, where the oscillation period was stable over a physiological range of temperatures from 24 °C to 30 °C (Q10 = 1.00-1.10). A Clock Tube Model developed could mimic hyphal growth observed in the serpentine chip and provides a mechanism for the stable banding patterns seen in race tubes at the macroscopic scale and synchronization through molecules riding the growth wave in the device.
摘要：
利用带有蛇形通道的微流控芯片，我们用带有粗糙神经孢子菌菌丝体的琼脂塞接种芯片，并成功捕获了通道中的单个菌丝。第一次,我们报告菌丝中存在一个自主时钟。每半小时沿菌丝同时测量由时钟控制的基因2启动子（ccg-2p）驱动的mCherry报告基因的荧光，持续至少6天。我们在很宽的时间范围内夹带了单个菌丝，包括6、12、24和36小时天。在单个蛇形通道中跟踪的菌丝高度同步（K=0.60-0.78）。此外，菌丝还显示了温度补偿特性，在24°C至30°C的生理温度范围内，振荡周期是稳定的（Q10=1.00-1.10）。开发的时钟管模型可以模拟蛇形芯片中观察到的菌丝生长，并提供了一种机制，用于在宏观尺度上在种族管中看到的稳定条带模式，并通过分子在设备中利用生长波实现同步。
