Biochemical and Molecular Biological Assessments of Neonatal HypoxiaIschemia: Cell Signaling

Neonatal hypoxia–ischemia (HI) induces a series of intracellular signaling events, including destructive and protective mechanisms. The destructive events include neuronal membrane depolarization, excitotoxicity, free radical injury, overactivation of calcium-dependent enzymes like calpains, proapoptotic protein release from mitochondria, and caspase activation. The protective events include upregulation of trophic factors and stimulation of neural stem cell proliferation. The injury after HI depends on the balance between destructive and protective events. Characterizing the mechanisms of injury and repair is important to be able to devise protective treatments or effective rehabilitation strategies. These mechanisms can be detected by analyzing RNA, protein level, or enzyme activity by using PCR, microarrays, ELISA, western blotting, or fluorometric assays of enzyme substrate turnover. This chapter illustrates how HI-related events can be studied using these methods.