摘要：

Elucidation of cellular and molecular mechanisms of excitotoxic neuronal injury have been facilitated by the development of several cell biological technologies—namely, (1) neural cell culture, (2) quantitative assays of neuronal injury and death, (3) fluorescence imaging of intracellular free calcium levels ([Ca 2+ ] i ), (4) evaluation of mitochondrial function, (5) quantification of reactive oxygen species, (6) assessment of cytoskeletal alterations elicited by excitotoxic/metabolic insults, and (7) detection of "stress response" proteins. This chapter presents protocols for each of these technical approaches, as applied to embryonic rat and human brain neurons in dissociated cell culture. One strategy employed to study neuronal death is to elucidate the mechanisms that normally protect neurons from adverse environmental conditions. These studies have shown that many cellular signaling mechanisms exist that are designed to protect neurons against adverse environmental conditions such as metabolic and excitotoxic insults. Many of the neuroprotective strategies acquired during evolution involve systems that control calcium and free radical metabolism.

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