We have determined the endogenous levels of neuropeptide Y-like immunoreactivity (NPY-LI) in retinal extracts from pigs, cats, rabbits, chickens, and frogs by radioimmunoassay (RIA). The NPY-LI levels varied among the species. The highest concentration was found in frog retina, where seasonal variations were seen, 861 +/- 31 pmol g(-1) wet weight in the autumn and 334 +/- 26 pmol g(-1) wet weight in the spring. Lower levels were demonstrated in chicken and pig retina, 4.1 +/- 0.4 and 3.6 +/- 0.3 pmol g(-1) wet weight, respectively. The lowest concentration was demonstrated in rabbit retina, 2.0 +/- 0.3 pmol g(-1) wet weight. (All values are expressed as mean +/- S.E.M.). The NPY-LI in pig, rabbit, chicken and frog retina was characterized by high-performance liquid chromatography (HPLC). The main part of the extracted NPY-LI had an elution volume close to that to the porcine NPY. We have also analysed the evoked release of endogenous NPY from frog retina, induced either with light flashes (3 Hz, 300 lx), or with potassium depolarization of the neurons (40 mM). Light flashes and potassium induced an increased release of NPY-LI of 61 and 77%, respectively. NPY-LI in the efflux had the same HPLC retention time as that extracted directly from the retina.