Davies Brandon M, Katayama Jenna K, Monsivais Joshua E, Adams James R, Dilts Miriam E, Eberting Arielle L, Hansen Jason M
Cell Biology and Physiology Department, Brigham Young University, Provo, UT 84602, USA.
Cell Biology and Physiology Department, Brigham Young University, Provo, UT 84602, USA.
Biochim Biophys Acta Gen Subj. 2023 May;1867(5):130321. doi: 10.1016/j.bbagen.2023.130321. Epub 2023 Mar 3.
Glutathione (GSH) is the most abundant, small biothiol antioxidant. GSH redox state (E) supports developmental processes, yet with disrupted GSH E, poor developmental outcomes may occur. The role of subcellular, compartmentalized redox environments in the context of redox regulation of differentiation is not well understood. Here, using the P19 neurogenesis model of cellular differentiation, kinetics of subcellular HO availability and GSH E were evaluated following oxidant exposure.
Stably transfected P19 cell lines expressing HO availability or GSH E sensors, Orp1-roGFP or Grx1-roGFP, respectively, targeted to the cytosol, mitochondria, or nucleus were used. Dynamic, compartmentalized changes in HO availability and GSH E were measured via spectrophotometric and confocal microscopy over 120 min following treatment with HO (100 μM) in both differentiated and undifferentiated cells.
Generally, treated undifferentiated cells showed a greater degree and duration of both HO availability and GSH E disruption than differentiated neurons. In treated undifferentiated cells, HO availability was similar in all compartments. Interestingly, in treated undifferentiated cells, mitochondrial GSH E was most affected in both the initial oxidation and the rebound kinetics compared to other compartments. Pretreatment with an Nrf2 inducer prevented HO-induced effects in all compartments of undifferentiated cells.
Disruption of redox-sensitive developmental pathways is likely stage specific, where cells that are less differentiated and/or are actively differentiating are most affected.
Undifferentiated cells are more susceptible to oxidant-induced redox dysregulation but are protected by chemicals that induce Nrf2. This may preserve developmental programs and diminish the potential for poor developmental outcomes.
谷胱甘肽（GSH）是最丰富的小型生物硫醇抗氧化剂。谷胱甘肽氧化还原状态（E）支持发育过程，但谷胱甘肽E受到破坏时，可能会出现不良发育结果。在分化的氧化还原调节背景下，亚细胞分隔的氧化还原环境的作用尚不清楚。在这里，使用细胞分化的P19神经发生模型，评估了氧化剂暴露后亚细胞HO可用性和谷胱甘肽E的动力学。
使用稳定转染的P19细胞系，分别表达靶向细胞质、线粒体或细胞核的HO可用性或谷胱甘肽E传感器，即Orp1-roGFP或Grx1-roGFP。在用HO（100μM）处理分化和未分化细胞后的120分钟内，通过分光光度法和共聚焦显微镜测量HO可用性和谷胱甘肽E的动态、分隔变化。
一般来说，处理后的未分化细胞比分化的神经元表现出更大程度和更长时间的HO可用性和谷胱甘肽E破坏。在处理后的未分化细胞中，所有区室的HO可用性相似。有趣的是，在处理后的未分化细胞中，与其他区室相比，线粒体谷胱甘肽E在初始氧化和反弹动力学中受影响最大。用Nrf2诱导剂预处理可防止HO对未分化细胞所有区室的影响。
氧化还原敏感的发育途径的破坏可能是阶段特异性的，其中分化程度较低和/或正在积极分化的细胞受影响最大。
未分化细胞更容易受到氧化剂诱导的氧化还原失调的影响，但受到诱导Nrf2的化学物质的保护。这可能会保留发育程序并降低不良发育结果的可能性。
