Cervený Jan, Nedbal Ladislav
Institute of Systems Biology and Ecology, Academy of Sciences CR, Nové Hrady, Czech Republic.
J Biol Rhythms. 2009 Aug;24(4):295-303. doi: 10.1177/0748730409338367.
These experiments aim to reveal the dynamic features that occur during the metabolism of the unicellular, nitrogen fixing cyanobacterium Cyanothece sp. when exposed to diverse circadian forcing patterns (LD 16:8, LD 12:12, LD 8:16, LD 6:6). The chlorophyll concentration grew rapidly from subjective morning when first illuminated to around noon, then remained stable from later in the afternoon and throughout the night. The optical density measured at 735 nm was stable during the morning chlorophyll accumulation, then increased in the early afternoon toward a peak, followed at dusk by a rapid decline toward the late night steady state. The authors propose that these dynamics largely reflect accumulation and subsequent consumption of glycogen granules. This hypothesis is consistent with the sharp peak of respiration that coincides with the putative hydrocarbon catabolism. In the long-day regimen (LD 16:8), these events may mark the transition from the aerobic photosynthetic metabolism to microaerobic nitrogen metabolism that occurs at dusk, and thus cannot be triggered by the darkness that comes later. Rather, control is likely to originate in the circadian clock signaling an approaching night. To explore the dynamics of the link between respiration and circadian oscillations, the authors extrapolated an earlier model of the KaiABC oscillator from Synechococcus elongatus to Cyanothece sp. The measured peak of respiratory activity at dusk correlated strongly in its timing and time width with the modeled peak in accumulation of the KaiB(4) complex, which marks the late afternoon phase of the circadian clock. The authors propose a hypothesis that high levels of KaiB(4) (or of its Cyanothece sp. analog) trigger the glycogen catabolism that is reflected in the experiments in the respiratory peak. The degree of the correlation between the modeled KaiB(4) dynamics and the dynamics of experimentally measured peaks of respiratory activity was further tested during the half-circadian regimen (LD 6:6). The model predicted an irregular pattern of the KaiABC oscillator, quite unlike mechanical or electrical clock pacemakers that are strongly damped when driven at double their endogenous frequency. This highly unusual dynamic pattern was confirmed experimentally, supporting strongly the validity of the circadian model and of the proposed direct link to respiration.
这些实验旨在揭示单细胞固氮蓝藻蓝细菌在暴露于不同昼夜节律强迫模式(LD 16:8、LD 12:12、LD 8:16、LD 6:6)时新陈代谢过程中出现的动态特征。叶绿素浓度从首次光照后的主观早晨迅速增长至中午左右,然后从下午晚些时候到整个夜间保持稳定。在735 nm处测得的光密度在早晨叶绿素积累期间稳定,然后在下午早些时候增加至峰值,黄昏时迅速下降至深夜稳定状态。作者提出这些动态变化在很大程度上反映了糖原颗粒的积累和随后的消耗。这一假设与与假定的碳氢化合物分解代谢同时出现的呼吸尖峰一致。在长日照方案(LD 16:8)中,这些事件可能标志着在黄昏时从有氧光合代谢向微需氧氮代谢的转变,因此不能由随后到来的黑暗触发。相反,控制可能源于昼夜节律时钟发出夜晚即将来临的信号。为了探索呼吸与昼夜节律振荡之间联系的动态变化,作者将来自聚球藻的KaiABC振荡器的早期模型外推至蓝细菌。在黄昏时测得的呼吸活动峰值在时间和时间宽度上与模拟的KaiB(4)复合物积累峰值密切相关,该峰值标志着昼夜节律时钟的下午晚些时候阶段。作者提出了一个假设,即高水平的KaiB(4)(或其蓝细菌类似物)触发了糖原分解代谢,这在呼吸峰值实验中得到了体现。在半日节律方案(LD 6:6)期间,进一步测试了模拟的KaiB(4)动态变化与实验测量的呼吸活动峰值动态变化之间的相关程度。该模型预测了KaiABC振荡器的不规则模式,这与机械或电子时钟起搏器在以其内源频率的两倍驱动时会强烈衰减的情况截然不同。这一高度不寻常的动态模式通过实验得到了证实,有力地支持了昼夜节律模型以及所提出的与呼吸的直接联系的有效性。