Action potentials of inhibitory burst neurons (IBNs) were extracellularly recorded in the pontomedullary reticular formation in the cat. These neurons were identified by their burst activity coincident with the quick inhibitory phase of the contralateral abducens nerve during vestibular nystagmus and by their antidromic activation from the contralateral abducens nucleus. During extracellular recording from the soma of single IBNs, another electrode for microstimulation was systematically tracked throughout the brain stem. For each IBN investigated, the effective sites for antidromic activation were invariably found in the contralateral abducens, prepositus hypoglossi, medial vestibular nuclei and the area ventral to the prepositus hypoglossi nucleus. Stimulation of neither the ipsilateral brain stem nor the oculomotor nuclei evoked antidromic responses in IBNs. Extracellular spikes of single IBNs and neurons in the overlying projection area were recorded simultaneously. Their correlation was examined by using peri-spike time histograms. Shortly after the spikes of single IBNs, the activity of motoneurons and internuclear interneurons in the abducens nucleus, and of type II neurons in the prepositus hypoglossi and vestibular nuclei, was depressed. Connections of IBNs with ipsilateral medial rectus motoneurons were studied by spike-triggered averaging of membrane potentials of the motoneurons and action potentials of the medial rectus nerve. Single IBN spikes induced a di- or polysynaptic disfacilitation in the motoneurons. This disfacilitation was concluded to be mediated by some of the above-described interneurons which were directly inhibited by IBNs. Their depressant effect on medial rectus motoneuronal spike activity was comparable to that on the spike activity of contralateral abducens motoneurons.