Fictive swimming activity was induced in isolated spinal cords of adult lampreys Ichthyomyzon unicuspis and Petromyzon marinus by addition of D-glutamate or N-methyl-D,L-aspartate (NMA) to the bathing fluid. Propriospinal interneurons are defined as nerve cells within the spinal cord with projections longer than 1 segment. The hypothesis that propriospinal interneurons contribute to intersegmental coordination during fictive swimming was tested using electrical stimulation, extracellular recording, and separated compartments. Stimulation of the split caudal end of the spinal cord indirectly excited ascending propriospinal interneurons, which enhanced and entrained bursts in rostral contralateral ventral roots. Indirect electrical stimulation of descending propriospinal interneurons could delay and diminish bursts in caudal contralateral ventral roots. Extracellular recordings from the rostral and caudal split ends of the spinal cord sometimes showed spike activities in phase with contralateral or ipsilateral ventral roots. Inhibition of 1-3 segments by spot applications of glycine or gamma-aminobutyric acid (GABA) did not interrupt normal coordination or rostrocaudal phase lag. When a middle region of spinal cord was inhibited in a compartment with GABA or glycine, the caudal spinal cord could entrain the bursts in rostral ventral roots. In a few preparations the caudal region induced antiphasic bursts in previously silent rostral roots through the inhibited region. The maximum separation for caudal-upon-rostral antiphasic entrainment was approximately 20 segments in Ichthyomyzon and 36 segments in Petromyzon. Increased concentrations of an excitatory amino acid in a rostral compartment could produce descending entrainment of bursts in an adjacent caudal compartment at a higher frequency with rostrocaudal phase lag. The rostral-upon-caudal entrainment could still occur through spot applications of GABA or glycine but not through long inhibited regions. Two hypothetical groups of propriospinal interneurons are proposed for the coordination of swimming activities in the isolated spinal cords of adult lampreys. 1) Crossed, ascending interneurons may be excited in phase with nearby motoneurons and may excite and entrain rostral pattern generators on the opposite side. 2) Short, commissural interneurons may be excited in phase with nearby motoneurons and may inhibit contralateral generators.