Methods for estimating the contribution of proenkephalin A and proenkephalin B input to neuronal areas.

Final expression of the proenkephalin A and proenkephalin B (prodynorphin) gene should yield a fixed ratio of enkephalin-like peptides depending on their precursor origin. If a tissue contained peptides only from proenkephalin A this ratio would be 4/1/1/1 (Met-Enk/Leu-Enk/Met-Enk-Arg6-Phe7/Met-Enk-Arg6-Gly7-Leu8). On the other hand, if the tissue-derived opiates were from prodynorphin, only Leu-enkephalin should be found. The techniques described here combining oxidation of the Met-Enk species, reverse-phase HPLC, and measurement with a radioimmunoassay which recognized many of the opiate-like peptides have been found useful for determining the precursor origin of enkephalins within different biological sources.