The effects of GABA-related compounds microinjected into the midbrain periaqueductal gray (PAG) on the tail-flick reflex (TF) and on the activity of tail-flick related neurons in the rostral ventromedial medulla (RVM), were studied in barbiturate anesthesized rats. Neurons whose activity either decreased (off-cells) or increased (on-cells) immediately prior to TF were examined. Bicuculline and picrotoxin microinjected into the ventrolateral aspect of the caudal PAG inhibited the TF, increased the spontaneous activity of the off-cells and decreased that of the on-cells. Concomitant with the increase in TF latency, the TF-related deceleration of the off-cells and acceleration of the on-cells were reduced. These effects were reversed by a microinjection of muscimol (MUS) into the PAG. The analgesic effect of morphine microinjected into the PAG was also reversed by a MUS microinjection at the same site. These results support the hypothesis that a GABAergic synapse inhibits cells in the PAG which modulate nociceptive transmission at the spinal level through actions on neurons in the RVM.