在肌萎缩侧索硬化症小鼠模型中氧化应激和缺氧应激的体内成像。
In Vivo Imaging of Oxidative and Hypoxic Stresses in Mice Model of Amyotrophic Lateral Sclerosis.
机构信息
Division of Neurology and Clinical Neuroscience, Department of Internal Medicine III, Yamagata University School of Medicine, Yamagata, Japan.
Department of Neurology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
出版信息
Methods Mol Biol. 2022;2525:289-294. doi: 10.1007/978-1-0716-2473-9_22.
Oxidative and hypoxic stresses are associated with the degeneration of both motor neurons and skeletal muscles in amyotrophic lateral sclerosis (ALS). In vivo bioluminescent imaging is used to monitor cellular responses to oxidative and hypoxic stresses in living ALS model mice bearing G93A-human Cu/Zn superoxide dismutase (SOD1) longitudinally using the IVIS spectrum imaging system. Double transgenic mice bearing both Keap1-dependent oxidative stress detector No-48 (OKD48) and G93A-SOD1 are useful for in vivo imaging of oxidative stress in ALS. We developed a bioluminescence resonance energy transfer (BRET) probe that is regulated by HIF-1α-specific ubiquitin-proteasome system. G93A-SOD1 mice injected with the BRET probe are useful to investigate the spatiotemporal responses to hypoxic stress in ALS. In this chapter, we introduce a practical protocol of in vivo imaging of both oxidative and hypoxic stress in ALS model mice.
氧化应激和缺氧应激与肌萎缩侧索硬化症（ALS）中运动神经元和骨骼肌的退化有关。在活体中，使用生物发光成像来监测携带 G93A-人类 Cu/Zn 超氧化物歧化酶（SOD1）的 ALS 模型小鼠对氧化应激和缺氧应激的细胞反应，使用 IVIS 光谱成像系统进行纵向监测。携带 Keap1 依赖性氧化应激探测器 No-48（OKD48）和 G93A-SOD1 的双转基因小鼠可用于 ALS 中氧化应激的活体成像。我们开发了一种受 HIF-1α 特异性泛素-蛋白酶体系统调节的生物发光共振能量转移（BRET）探针。用 BRET 探针注射的 G93A-SOD1 小鼠可用于研究 ALS 中缺氧应激的时空反应。在本章中，我们介绍了一种在 ALS 模型小鼠中进行氧化应激和缺氧应激活体成像的实用方案。
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