High levels of NPY in rabbit cerebrospinal fluid and immunohistochemical analysis of possible sources

We have analyzed rabbit cerebrospinal fluid for neuropeptide Y (NPY)-like immunoreactivity, using high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) and examined the anatomical relationship of NPY-containing fibers to the cerebral vasculature and the third cerebral ventricle. Cerebrospinal fluid (CSF) obtained from the cisterna magna of rabbits was injected into a C 18 column and subjected to HPLC. The fractions were collected, dried and reconstituted in buffer for NPY radioimmunoassay. A single peak of NPY immunoreactivity was obtained which corresponded in retention time to synthetic porcine NPY. Analysis of CSF samples produced displacement curves parallel to the standard curve. Immunohistochemistry revealed numerous NPY-labeled fibers which penetrated the ependymal lining of the third cerebral ventricle and directly bordered the ventricular lumen. Other fibers were observed in the pia which lines the ventral aspect of the hypothalamus. The basilar artery, its branches and other cerebral vessels were surrounded by NPY-labeled fibers. The results show that: (1) approximately 1 ng/ml of NPY immunoreactivity which corresponds chromatographically to synthetic porcine NPY is present in rabbit CSF; (2) NPY-containing fibers surround the basilar artery and other cerebral vessels; (3) NPY may be released into the CSF from axons in the pia and from axons which penetrate the ependymal lining of the third ventricle. These observations form the basis for our analysis of the vasoconstrictor effects of NPY and its role in cerebrovasospasm after experimental subarachnoid hemorrhage.