1. Field potentials and intracellular records were obtained from the medial rectus subdivision of the IIIrd nucleus in anesthetized cats following electrical stimulation of the abducens nuclei, vestibular nerves, pontomedullary brain stem, and the medial longitudinal fasciculi (MLF). 2. Stimulation of the contralateral abducens nucleus produced unique field potentials in the medial rectus subdivision. They consisted of an early sharp transient volley followed by a slower postsynaptic negativity. 3. Monosynaptic EPSPs were evoked in medial rectus motoneurons following contralateral abducens nucleus stimulation. The EPSP amplitudes were graded when the stimulus intensity was increased from threshold to supramaximal. EPSPs produced by contralateral abducens nucleus stimulation were larger in amplitude than those produced by ipsilateral vestibular nerve stimulation. The current-voltage relationship and reversal potentials for Vi- and abducens-evoked EPSPs were similar and indicated an overlapping location of excitatory synaptic terminals on medial rectus motoneurons. 4. Secondary vestibular axons activated monosynaptically by ipsilateral vestibular nerve stimulation were not recruited by abducens nucleus stimulation. 5. Ipsilateral MLF stimulation produced EPSPs with similar profiles as those observed following abducens nucleus stimulation; however, stimulation of the contralateral MLF at comparable stimulus intensities did not produce any changes in transmembrane potential. 6. When higher intensity stimuli were applied to the contralateral MLF, the synaptic potentials recorded in the medial rectus were occluded by those produced by weaker stimulation applied to the ipsilateral MLF. This suggests that the potentials resulting from stronger contralateral stimulation were due to current spread to the ipsilateral MLF. 7. While recording in the medial rectus subdivision, various sites in the ponto-medullary brain stem were explored with a stimulating electrode. Analysis of evoked field potentials suggested that the ascending internuclear axons were contained only in the MLF ipsilateral to the medial rectus. Acute brain stem lesions confirmed this suggestion. 8. Chronic lesions were placed in the brain stem to isolate the abducens nucleus from either extrinsic fibers of passage or axon collaterals. Acute electrophysiological experiments in these chronic animals corroborated the suggestion that the medial rectus pathway originated from within the abducens nucleus. 9. We conclude that axons from the internuclear neurons of the abducens nucleus exit from the nucleus medially, cross the midline, ascend in the opposite MLF, and terminate monosynaptically on medial rectus motoneurons. 10. we believe that the syndrome of internuclear ophthalmoplegia associated clinically with lesions of the medial longitudinal fasciculus could be due to the absence of ascending physiological activity from internuclear neurons of the abducens nucleus.