Single Cells of Neurospora Crassa Show Circadian Oscillations, Light Entrainment, Temperature Compensation, and Phase Synchronization
Single Cells of Neurospora Crassa Show Circadian Oscillations, Light Entrainment, Temperature Compensation, and Phase Synchronization
复制标题
DOI：
10.1109/access.2019.2910731
复制
发表时间：
2019-01-01
期刊：
影响因子：
3.9
通讯作者：
Arnold, Jonathan
中科院分区：
文献类型：
--
作者：
Deng, Zhaojie;Cheong, Jia Hwei;Arnold, Jonathan
使用微流体装置，在不同的光照和温度条件下，每半小时对超过 1000 个粗糙脉孢菌单细胞同时测量由时钟控制基因 2 启动子 (ccg-2p) 驱动的记录器 (mCherry 或 mVenus) 基因的荧光，持续 10 天。单细胞能够在很宽的日长范围内吸收光线，包括 6、12 或 36 小时。此外，荧光振荡周期在 20 摄氏度至 30 摄氏度的生理温度范围内保持非常稳定 (Q(10) = 1.00-1.07)。这些结果提供了粗糙猪笼草大多数单细胞中存在自主时钟的证据。虽然大多数细胞都有时钟，但通过相位测量，时钟之间存在很大差异，这就提出了一个问题：单细胞中的细胞时钟如何进行相位同步，以在 10(7) 细胞的宏观水平上实现真核系统中的昼夜节律行为，而大多数时钟测量都是在 10(7) 细胞的宏观水平上进行的。通过让一组细胞在熄灯前多接受 6 或 12 小时的光照 (D/D)，将单细胞异相放置。相对于两个非混合对照群体，混合群体的平均相位差减小。
Using a microfluidics device, fluorescence of a recorder (mCherry or mVenus) gene driven by a clock-controlled gene-2 promoter (ccg-2p) was measured simultaneously on over 1000 single cells of Neurospora crassa every half hour for 10 days under each of varied light and temperature conditions. Single cells were able to entrain to light over a wide range of day lengths, including 6, 12, or 36 h days. In addition, the period of oscillations in fluorescence remained remarkably stable over a physiological range of temperatures from 20 degrees C to 30 degrees C (Q(10) = 1.00-1.07). These results provide evidence of an autonomous clock in most single cells of N. crassa. While most cells had clocks, there was substantial variation between clocks as measured by their phase, raising the question of how such cellular clocks in single cells phase-synchronize to achieve circadian behavior in eukaryotic systems at the macroscopic level of 10(7) cells, where most measurements on the clock are performed. Single cells were placed out of phase by allowing one population to receive 6 or 12 h more light before lights out (D/D). The average phase difference was reduced in the mixed population relative to two unmixed control populations.