This study describes the distribution of peptide sequences derived from the prepro-vasoactive intestinal polypeptide (preproVIP) molecule in perivascular nerves of rat brain arteries and arterioles. The peptides were identified by immunohistochemistry using highly specific antibodies. Five peptide sequences (preproVIP 60–76, peptide histidine isoleucine (PHI), preproVIP 111–122, VIP, and preproVIP 156–170) were identified in the perivascular nerves throughout the arterial cerebral circulation. The density of the immunoreactive fibers was highest in the nerves of the larger extracerebral arteries, declining in smaller branching arteries. All peptide sequences were identified in the nerves of small pial arterioles overlying the cortical convexity, whereas capillaries and veins contained no immunoreactive material. Dendritic processes of neocortical neurons immunoreactive for VIP and PHI could be followed towards the brain surface where the processes penetrated into the pial layer, often close to the pial vasculature. Some of the processes were also observed to enter the Virchow-Robin space, close to the arterioles. It is possible that cortical nerve cells containing VIP and PHI release the peptides in the perivascular space during periods of activity and thereby contribute to local vasodilatation associated with changes of neuronal function.