One of the main characteristics of central sensitization associated with postinjury pain and chronic pain is increased excitability of the dorsal horn neurons in the spinal cord. Two electrophysiological features associated with the origin and modulation of central sensitization are wind-up of action potential frequency and long-term potentiation (LTP), which have been demonstrated previously in the intact dorsal horn. Here we present evidence for electrically evoked sensitization of dorsal horn neurons in a two-compartment cell culture system of rat dorsal root ganglia (DRGs) and dorsal horn neurons. Whole-cell recordings of dorsal horn neurons showed that repetitive low-frequency stimulation of DRG axons induced a frequency-dependent cumulative depolarization of the membrane potential with a concomitant increase in action potential frequency in a subset of neurons (41%). The characteristics presented here for dissociated cells are in accordance with those ascribed to classical wind-up in the intact dorsal horn. In addition, tetanic stimulation of DRG axons resulted in a significant increase in the number of action potentials in response to test stimuli in 42% of the cells tested. This prolonged potentiation of neuronal excitability in the dorsal horn lasted throughout the recording period (>1 hr) and tended to be voltage dependent in an LTP-like manner. To our knowledge, this is the first time that wind-up and LTP-like responses are reported for dorsal horn neurons in cell culture.