Qiao Xinhua, Kang Lu, Shi Chang, Ye Aojun, Wu Dongli, Huang Yuyunfei, Deng Minghao, Wang Jiarui, Zhao Yuzheng, Chen Chang
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
School of Basic Medical Sciences of Southwest Medical University, Luzhou, 646000, China.
Stress Biol. 2023 Jun 12;3(1):17. doi: 10.1007/s44154-023-00096-z.
Fasting is a popular dietary strategy because it grants numerous advantages, and redox regulation is one mechanism involved. However, the precise redox changes with respect to the redox species, organelles and tissues remain unclear, which hinders the understanding of the metabolic mechanism, and exploring the precision redox map under various dietary statuses is of great significance. Twelve redox-sensitive C. elegans strains stably expressing genetically encoded redox fluorescent probes (Hyperion sensing HO and Grx1-roGFP2 sensing GSH/GSSG) in three organelles (cytoplasm, mitochondria and endoplasmic reticulum (ER)) were constructed in two tissues (body wall muscle and neurons) and were confirmed to respond to redox challenge. The HO and GSSG/GSH redox changes in two tissues and three organelles were obtained by confocal microscopy during fasting, refeeding, and satiation. We found that under fasting condition, HO decreased in most compartments, except for an increase in mitochondria, while GSSG/GSH increased in the cytoplasm of body muscle and the ER of neurons. After refeeding, the redox changes in HO and GSSG/GSH caused by fasting were reversed in most organelles of the body wall muscle and neurons. In the satiated state, HO increased markedly in the cytoplasm, mitochondria and ER of muscle and the ER of neurons, while GSSG/GSH exhibited no change in most organelles of the two tissues except for an increase in the ER of muscle. Our study systematically and precisely presents the redox characteristics under different dietary states in living animals and provides a basis for further investigating the redox mechanism in metabolism and optimizing dietary guidance.
禁食是一种流行的饮食策略，因为它有诸多益处，氧化还原调节是其中涉及的一种机制。然而，关于氧化还原物种、细胞器和组织的精确氧化还原变化仍不清楚，这阻碍了对代谢机制的理解，探索不同饮食状态下的精确氧化还原图谱具有重要意义。我们构建了12种对氧化还原敏感的秀丽隐杆线虫品系，它们在两种组织（体壁肌肉和神经元）的三种细胞器（细胞质、线粒体和内质网（ER））中稳定表达基因编码的氧化还原荧光探针（用于检测HO的Hyperion和用于检测GSH/GSSG的Grx1-roGFP2），并证实其对氧化还原挑战有反应。通过共聚焦显微镜观察，我们获得了禁食、再喂食和饱腹状态下两种组织和三种细胞器中HO和GSSG/GSH的氧化还原变化。我们发现，在禁食条件下，除线粒体中HO增加外，大多数区室中的HO减少，而体壁肌肉细胞质和神经元内质网中的GSSG/GSH增加。再喂食后，体壁肌肉和神经元的大多数细胞器中，由禁食引起的HO和GSSG/GSH的氧化还原变化发生逆转。在饱腹状态下，肌肉的细胞质、线粒体和内质网以及神经元的内质网中的HO显著增加，而在这两种组织的大多数细胞器中，GSSG/GSH没有变化，除了肌肉内质网中的GSSG/GSH增加。我们的研究系统而精确地呈现了活体动物在不同饮食状态下的氧化还原特征，为进一步研究代谢中的氧化还原机制和优化饮食指导提供了依据。
