In Vivo Imaging of Oxidative and Hypoxic Stresses in Mice Model of Amyotrophic Lateral Sclerosis

Abstract

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.

Keywords: Amyotrophic lateral sclerosis (ALS); Bioluminescence resonance energy transfer (BRET); Hypoxia inducible factor-1 (HIF-1); Hypoxic stress; In vivo imaging; Nuclear erythroid 2-related factor 2 (Nrf2); Oxidative stress; Superoxide dismutase (SOD1).