Isolation and Culture of Sensory Neurons From the Dorsal-Root Ganglia of Embryonic or Adult Rats
There is increasing use of isolated sensory neuronal preparations to examine the cellular mechanisms involved in pain signaling. Indeed, these in viro preparations have several advantages that make them beneficial for examining physiological and/or pathological processes affecting neuronal function. With isolated cells it can be determined whether various inflammatory mediators and algogenic agents have direct actions on sensory neurons. Additionally, the intracellular signaling pathways for agents that modulate the excitability and sensitization of sensory neurons can be examined. Finally, the concentrations of mediators and drugs that are used to alter cell function can be well controlled. The purpose of this chapter is to provide the reader with detailed methods for the harvest and growth of embryonic and adult rat sensory neurons (dorsal root ganglia neurons) in culture. Because numerous methods for growing sensory neurons exist, the rationale for certain aspects of the protocols described in the chapter are included, as are discussions of potential pitfalls.
Key WordsCell dissociation cell culture dorsal-root ganglia nerve growth factor rat sensory neurons growth medium
- 12.Martin, D. P., Wallace, T. L., and Johnson, E. M. Jr. (1990) Cytosine arabinoside kills postmitotic neurons in a fashion resembling trophic factor deprivation: evidence that a deoxycytidine-dependent process may be required for nerve growth factor signal transduction. J. Neurosci. 10, 184–193.PubMedGoogle Scholar