Novel paired ependymal ridges (EPRS) have been identified in the ventrolateral wall of the cerebral aqueduct of the rat. Scanning electron microscopic techniques revealed that the dorsolateral and ventromedial ridges differ from each other in surface morphology. Using the PAP immunohistochemical technique, the EPRS demonstrate a dense innervation of both serotonin and LH-RH fibers when compared to the surrounding midbrain periaqueductal gray and the 'non-ridge' ependyma of the aqueduct. Each ridge is composed of a single layer of ependymal cells with a central core of subependymal cells and numerous blood vessels. Ependymal cells resembling tanycytes extend between the lumen of the aqueduct and the subependymal capillaries. It is suggested that the EPRS may serve as sites for the release of neurochemicals into the cerebrospinal fluid.