Retrograde dye staining, combined with histochemical and immunohistochemical techniques, were used to characterize penile neurons in the major pelvic ganglion of the rat. Of the penile neurons 92% were immunoreactive for vasoactive intestinal polypeptide, while 95% of penile neurons stained intensely for acetylcholinesterase. None of the neurons were immunoreactive for tyrosine hydroxylase. Penile neurons in the pelvic plexus receive preganglionic input from the pelvic and the hypogastric nerve, yet the shared histochemical features of the postganglionic neurons suggest that the two pathways have a similar role in penile erectile tissue.