A Calcium Imaging Approach to Measure Functional Sensitivity of Neurons

Pain associated with chemotherapy and radiation therapy is one of the most common reasons for discontinuation of these treatments and has a dramatic effect on the quality of life in cancer patients. However, the mechanisms underlying chemotherapy and radiation therapy associated with pain are not well understood. Pain sensations are mediated through sensory neurons whose cell bodies are located in the dorsal root ganglia (DRG). Pain mediators activate these sensory neurons causing an influx of ions, including calcium. One common technique to study pain is to use primary cell culturing mouse DRG to study this calcium influx in vitro. This protocol details from an isolation to culture and maintenance of DRG neurons and functional recording using calcium imaging caused by either pain mediators or neuronal sensitization that are induced by drugs that are often used in the treatment of cancer.