The present study was designated to determine the origin of commissural axons in the hippocampus. One hippocampus of 94 rats was pressure injected with 40% horseradish peroxidase (Sigma VI), or with 2-4% wheat germ agglutinin HRP (E-Y Labs). Injections (0.001 to 0.1 microliter) were made through glass micropipettes with fitted plungers. Pipettes were positioned stereotaxically, and by electrophysiological monitoring through the injection syringe. An ipsilateral stimulating electrode activated CA3 and CA1 cells via Schaffer collaterals. Population potentials were monitored as the recording pipette was advanced from the cortical surface into the hippocampus. Wave forms of monosynaptically elicited field potentials provided an accurate indicator of its position. Following survival periods of 24 hours, the brains were processed according to the Mesulam method. Forty-micron sections were serially mounted and counterstained. Injection sites and filled cells were plotted manually on a standard set of coronal sections. Our results indicate that field CA1 receives input from contralateral subfields CA1a and c, as well as from all CA3 subfields. In addition, rostral CA1 injections resulted in labeling of cells in the contralateral subiculum and entorhinal cortex. Homotopic connections exist between subfields CA3a and b; it appears that a major input to CA3c is from the contralateral polymorph cells of the dentate hilus. Commissural input to the dentate granule cells appears to be the giant polymorph and CA3c cells of the contralateral dentate hilus. With respect to the question of homotopicity, our results suggest that commissural connections are predominantly homotopic in the mediolateral plane, although CA1 and CA3 injections also resulted in contralateral labeling of hippocampal cells caudal to the levels of injection.