Whole-cell recording in isolated primary sensory neurons

The isolated sensory neuron in vitro is a powerful model with which to address a number of important neurobiological questions. Isolated neurons are relatively easy to prepare from both neonatal and adult animals and can be studied both acutely and after considerable time on culture. Intracellular recording is one of the most powerful ways to study these neurons. Methods are described for both the preparation of isolated sensory neurons in vitro as well as for recording major classes of ionic currents (Na(+), K(+), and Ca(2+)) from these neurons with whole cell voltage-clamp techniques. Methods are also provided for an initial characterization of active and passive electrophysiological properties of these neurons in current clamp as well as the use of perforated patch recording as a means to mitigate some of the limitations associated with conventional whole cell patch recording. The reader should be aware that the regulation of ion channels in sensory neurons may very subtle, requiring considerably more sophisticated protocols than have been provided here. The reader should also be aware that there is a tremendous heterogeneity among sensory neurons, which is both a curse and a blessing for those who wish to study them. Thus, the methods provided here should only be considered the starting point for a more detailed analysis of sensory neurons.