In a previous study we investigated the intraspecific defensive reactions evoked by excitation of neurons in the intermediate third of the midbrain periaqueductal gray matter (PAG) of the rat. Experiments revealed that activation of neurons in this region of the PAG mediated: (i) backward defensive behavior, characterized by upright postures and backward movements, and (ii) reactive immobility (“freezing”), in which the rat remained immobile, but reacted with backward defensive behavior to investigative, non-aggressive contact initiated by the partner. In the present study, we aimed to extend our understanding of PAG mediation of defensive behavior by observing: (i) in a non-aggressive social interaction test, the behavioral effects of microinjections of low doses of kainic acid (40 pmol in 200 nl) made in the caudal third of the PAG; and (ii) the behavioral and cardiovascular effects of microinjections of d, l-homocysteic acid (5–10 nmol in 50–100 nl) made in the PAG of the unanesthetized decerebrate rat. Kainic acid injections into the area lateral to the midbrain aqueduct in the caudal third of the PAG evoked: (i) forward avoidance behavior, characterized by forward locomotion and occasional hop/jumps; (ii) reactive immobility (“freezing”), in which the rat remained immobile, but reacted with forward avoidance behavior to investigative, non-aggressive contact initiated by the partner; and (iii) 22–28 kHz ultrasonic vocalizations. These injections also evoked a dramatic increase in defensive responsiveness to tactile stimuli on the half of the body contralateral, but not ipsilateral, to the site of injection. Electroencephalographic measurements indicated that none of these effects were secondary to seizure activity. In the decerebrate rat, d, l-homocysteic acid injections in the caudal third of the PAG evoked forward running movements along with increased blood pressure and heart rate, the strongest effects being evoked from the region lateral to the midbrain aqueduct. More rostrally, sites in the intermediate PAG evoked backward “defensive” movements, which were also associated with increased blood pressure and heart rate. These data, along with those from our previous study in the rat indicate that: (i) defensive reactions are integrated within a longitudinal neuronal column which spans the caudal two thirds of the lateral PAG; (ii) the lateral PAG “defensive behavior” column contains two distinct populations of neurons, one within the intermediate lateral PAG which integrates defensive behavior characterized by facing towards and backing away from a “threatening” stimulus, and a second in the caudal lateral PAG which integrates defensive behavior characterized by forward avoidance behavior; and (iii) neurons within the lateral PAG couple strong cardiovascular changes with each distinctive defensive behavior pattern. The emerging view from this and recent studies of this midbrain region in other species, suggests that similar rostrocaudal differences within a longitudinally oriented lateral PAG neuronal column represent a fundamental principle underlying the PAG organization of defensive behavior.