Direct projections from the medial preoptic area (MPO) to the pontine micturition center neurons directly projecting to the lumbosacral spinal cord were revealed electron microscopically in the rat by a double labeling method. Biotinylated dextran amine (BDA) was injected into the MPO and horseradish peroxidase (HRP) was injected into the lumbosacral cord segments. At light microscopic level, BDA-labeled presumptive axon terminals completely overlapped with HRP-labeled neurons in Barrington's nucleus. Electron microscopic observation showed that some BDA-labeled axon terminals made synaptic contacts with dendrites of HRP-labeled neurons in Barrington's nucleus. The present results indicated that the MPO may be involved in the modulation of the pontine micturition reflex in the rat.