Immunohistochemical localization of unique enkephalin sequences contained in preproenkephalin A in the guinea pig cochlea

The guinea pig cochlea was studied for the presence of immunoreactivities to the four unique enkephalin sequences contained in the preproenkephalin A. The antisera to Met-enkephalin-Arg6-Phe7 and Met-enkephalin-Arg6-Gly7-Leu8 were used as highly specific markers for the preproenkephalin A. Contrary to Met-enkephalin and Leu-enkephalin, also included in the present study, these two peptide sequences are not contained in pro-opiomelanocortin or preproenkephalin B (prodynorphin). All the four different antisera showed identical localization for the four peptide sequences, suggesting their coexistence in the same nervous pathway. Specific immunofluorescence was found in intraganglionic spiral bundle, inner spiral bundle, tunnel spiral bundle and in association with inner hair cells. The nerve fibers were thin and varicose, suggesting that most, if not all, of them were unmyelinated. Their localization indicates that the unique enkephalin sequences contained in preproenkephalin A are present in the cochlear efferent pathway to the inner hair cell region.