A derivative of the neural crest, sympathetic neurons have been utilized in both in vivo and in vitro studies to approach a number of basic questions concerning the development and function of the nervous system. The superior cervical ganglia (SCG), and particularly the sympathetic chain, can provide significant numbers of neurons with relatively little effort in dissection, and, with a known and available growth factor (nerve growth factor [NGF]), can be maintained in culture for prolonged periods of time. As a result, cultures of sympathetic neurons, many times from rats or mice, but also from avian and amphibian sources, have been utilized to study factors influencing synaptic function, neurotransmitter differentiation, neurotrophic dependence, dendritic development, and axonal growth, including the structure and function of growth cones. Studies of axonal elongation, substrate requirements, and molecular interactions, underlying neurite extension and ensheathment, have utilized explants; other experiments on dendrite growth, cell death, and neurotransmitter changes have used dissociated, even isolated, single neurons, depending on the question being asked. The following paragraphs contain detailed methods for the dissection of sympathetic neurons from embryonic, perinatal, and adult rats.