The neurocircuitry underlying regulation of bladder and distal colon function by Barrington's nucleus (the pontine micturition centre) was investigated in rats by identifying neurons which were transsynaptically labelled from these viscera, with pseudorabies virus (PRV) or genetically modified forms of PRV [PRV–β-galactosidase (PRV-β-Gal) and PRV–green fluorescent protein (PRV-GFP)]. PRV injection into the bladder or the colon of separate rats suggested an overlap in the distribution of bladder- and colon-related neurons in Barrington's nucleus, as well as a topographical arrangement whereby dorsal neurons were bladder-related and ventral neurons were colon-related. In rats injected with PRV-β-Gal into one viscera and PRV-GFP into another, neurons in the major pelvic ganglion and lumbosacral spinal cord were primarily single-labelled at relatively early survival times. With longer survival times many double-labelled neurons (>70%) appeared in Barrington's nucleus, suggesting that individual Barrington's nucleus neurons are synaptically linked to preganglionic parasympathetic neurons which independently innervate the colon or the bladder. In addition to these dual-labelled neurons, Barrington's nucleus neurons which were single-labelled from either viscera were observed and these exhibited a viscerotopic organization consistent with the single-labelling studies. Together, these findings suggest the existence of three neuronal populations in Barrington's nucleus, one which is synaptically linked to both the bladder and the colon and the other two populations which are specifically linked to either viscera. These anatomical substrates may underlie the central coordination of bladder and colon function and play a role in disorders characterized by a coexistence of bladder and colonic symptoms.