Although rhythmic behavior of mammalian spinal ventral horn networks has been extensively studied little is known about oscillogenesis in the spinal dorsal horn. The aims of this in vitro study were to record and determine the underlying mechanisms of potassium-evoked network field oscillations in the substantia gelatinosa of the neonatal rat dorsal horn, a lamina involved in nociceptive processing. Transient pressure ejection of a potassium solution evoked reproducible rhythmic activity in discrete areas of the substantia gelatinosa which lasted for 5-15 s with a single prominent peak in the 4-12 Hz frequency band (7.7 ± 0.1 Hz, n=60). Oscillations of similar frequency and amplitude were also observed in isolated dorsal horn quadrants. Application of CNQX (10 µM) reduced peak power amplitude and integrated power area (from 4-12 Hz) of the power spectrum, whereas D-AP5 (50 µM) had no effect on the potassium-evoked rhythm. Bicuculline (30 & [mu]M) or strychinine (10 µM) reduced the power amplitude and area. On combination of bicuculline (30 & [mu]M) and strychinine (10 µM) the reductions in power amplitude and area were not significantly different (P > 0.05) when compared to application of either drug alone. The gap junction blockers, carbenoxolone (100 µM) or octanol (1 mM) significantly reduced power amplitude and area. Although TTX (1 µM) or a calcium-free perfusate both caused reductions in the power amplitude and area, potassium-evoked rhythmic activity persisted. However, this persistent rhythm was further reduced on combination of calcium-free perfusate with octanol (1 mM) and was abolished using a cocktail of drugs. Blockade of the potassium delayed rectifier current by tetraethylammonium (5 mM) or the hyperpolarisation- activated current (I_h) by ZD7288 (10 µM) disrupted the synchronization of the potassium-induced oscillation. The frequency of potassium-induced rhythms was unaffected by any of the drugs tested. These novel findings demonstrate that transient pressure ejection of potassium evokes oscillatory activity in the substantia gelatinosa in vitro. This rhythm is partly dependent upon various receptors (AMPA/kainate, GABAA and glycine), ion channels (potassium delayed rectifier and I_h) and gap junctions. Oscillatory behaviour in the substantia gelatinosa could potentially play a role in the processing of nociceptive signals.