MET-ENKEPHALIN-ARG-GLY-LEU-LIKE IMMUNOREACTIVITY IN ADRENAL CHROMAEFIN CELLS AND CAROTID BODY CHIEF CELLS OF THE DOG AND MONKEY

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Abstract
Met-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg-Gly-Leu)-like immunoreactivity is shown in adrenal chromaffin cells and carotid body chief cells of the dog and monkey by immunohistochemistry using rabbit antiserum (R-0171) raised against synthetic Met-Enk-Arg-Gly-Leu conjugated with ascaris protein by the glutaraldehyde method. Immunoreactive sites were detected at the light microscopic level by the peroxidase antiperoxidase method and at the EM level by the protein A-colloidal Au method. Positive immunostaining was detected in both adrenal chromaffin cells and carotid body chief cells. The immunostaining was absorbed by preincubation of the antiserum with Met-Enk-Arg-Gly-Leu (0.1 .times. .**GRAPHIC**. g/l; dilution of the antiserum, 1:4000), while no absorption was observed with Leu- or Met-enkephalin (50 mg/l; dilution of the antiserum, 1:4000). The present immunostaining using the R-0171 antiserum was specific to the C-terminal region of the Met-Enk-Arg-Gly-Leu molecule. In EM, most of the immunoreactivity was demonstrated in the secretory granules. One or more Met-Enk-Arg-Gly-Leu-related peptides are contained in the carotid body as well as in the adrenal medulla.