Radioimmunoassays for fish tail neuropeptides: II. Development of a specific and sensitive assay for and the occurrence of immunoreactive urotensin II in the central nervous system and blood of Catostomus commersoni.

A sensitive radioimmunoassay (RIA) based on an antiserum to synthetic Gillichthys mirabilis urotensin II (UII) generated in rabbits, reacting with all known forms of the UII peptides, was developed. The UII was iodinated by either the chloramine-T or the lodogen method and was purified by high-pressure liquid chromatography. The antiserum, at a final dilution of 1:125,000, gave 50% binding of the iodinated UII. The sensitivity of the RIA was 1.8 +/- 0.2 pg/assay tube (1.2 +/- 0.2 fmol/tube; n = 7). Crossreactivity studies with various UII peptides, modified UII peptides, and fragments indicated that the immunoreactive recognition site of this antiserum is directed to the disulfide ring region (positions 6-11) of the UII peptides. Somatostatin 14, which has the part sequence 7-9 in common with UII peptides, did not crossreact. No detectable crossreactivity with urotensin I, urophysin B or D from Catostomus commersoni, arginine vasopressin, or arginine vasotocin could be shown. Displacement curves obtained with standard extracts of G. mirabilus urophyses were similar to those of synthetic G. mirabilus UII. comparison of RIA and bioassay of G. mirabilis urophysial extracts showed good correlation. Immunoreactive UII was detected in different parts of the brain, spinal cord, and blood of C. commersoni.