Characterization and location of Met5-enkephalin-arg6-phe7 stored in various rat brain regions

A specific antiserum against met5-enkephalin-arg6-phe7 was raised and used to study the distribution and characterization of met5-enkephalin-arg6-phe7-like immunoreactive material in rat brains by radioimmunoassay and immunohistochemical procedures. The antiserum appears to be directed to the COOH-terminus of the peptide, as it fails to cross-react with met5-enkephalin, met5-enkephalin-arg6, met5-enkephalin-arg6-arg7, met6-enkephalin-lys6, and leu-enkephalin. However, it cross-reacts with phe-met-arg-phe by about 10% and with phe-met-arg-phe-NH2 to an insignificant degree. The highest content of met5-enkephalin-arg6-phe7 was found in the striatum, which contains a dense network of immunoreactive varicose fibers and terminals, as well as immunoreactive cell bodies. The met5-enkephalin-arg6-phe7 in striatum can be released in a Ca2+-dependent manner by a depolarizing concentration of KCl, raising the possibility of a neuroregulatory role for met5-enkephalin-arg6-phe7. Characterization of the immunoreactive material by gel filtration and high pressure liquid chromatography revealed the presence of multiple forms of immunoreactive material in some brain regions.