The perineurial diffusion barrier to horseradish peroxidase (HRP) and ferritin was investigated in superior cervical ganglia of rats and mice. The ganglion was surrounded by a delicate epineurium and 2-5 perineurial lamellae joined by zonulae occludentes and desmosomes. Following local application of tracers the animals were killed after 5, 30, and 60 min and the distribution of HRP and ferritin was studied by light and electron microscopy. The inner layers of the ganglionic perineurium prevented diffusion of both HRP and ferritin perineurial lamellae investing the ganglion. HRP had often extended to the innermost lamella 60 min after application. HRP and ferritin were present in vesicles of ganglionic perineurial cells. There was no passage of tracers via intercellular junctions.