Behavioral methods for inferring anatomical linkage between rewarding brain stimulation sites.

Rats lever pressed for concurrent electrical stimulation of the lateral hypothalamus and ventral tegmentum. The pulse-pair stimulation technique was used, with the first pulse of each pair applied to one electrode and the second to the other electrode; the intrapair interval was varied. The effectiveness of stimulation, measured behaviorally, increased abruptly (within .4 msec) as the intrapair interval was increased in the range from 1.0 to 2.0 msec. These results, which do not resemble single-electrode refractory period results, are interpreted as evidence of collision in the directly stimulated, reward-related neurons linking the two sites. We conclude that self-stimulation of the medial forebrain bundle involves the direct activation of long-axon, longitudinal pathways. Estimates of the conduction velocity in the fibers subserving the collision-like effects are consistent with the properties of small myelinated axons but not central monoaminergic fibers.