Co-storage of adrenaline and noradrenaline with Met-enkephalin-Arg6-Gly7-Leu8 and Met-enkephalin-Arg6-Phe7 in chromaffin cells of hamster adrenal medulla.

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Abstract
The co-existence of catecholamines with opioid peptides in adult hamster adrenal chromaffin cells was studied by Sternberger''s peroxidase-antiperoxidase (PAP) method. Paraffin embedded serial sections (3-4 .mu.m thick) of adrenal glands perfusion-fixed with either Bouin''s fluid or 4% paraformaldehyde solution were immunostained using anti-adrenaline [epinephrine, E] (AP16-2), noradrenaline [norepinephrine, NE] (NAP1-18), -Met-enkephalin-Arg6-Gly7-Leu8 (Met-Enk-Arg-Gly-Leu:R-0171, NE2-206) and -Met-enkephalin-Arg6-Phe7 (Met-Enk-Arg-Phe: AP3-311) sera. Selective and specific immunostaining for E and NE was obtained in tissues fixed in 4% paraformaldehyde solution. Bouin''s fluid gave negative or unsatifactory results. Immunostaining with 2 kinds of Met-Enk-Arg-Gly-Leu (R-0171, NE2-206) and 1 kind of Met-Enk-Arg-Phe sera (AP3-311) was dependent upon the method of fixation used. The immunostaining in the Bouin-fixed tissues was stronger than that in the 4% paraformaldehyde-fixed ones. Secretory granules in chromaffin cells were responsible for both A- and NE-like and Met-Enk-Arg-Gly-Leu- and Met-Enk-Arg-Phe-like immunoreactivities. All E-storing (E) and NE-storing (NE) cells showed intense Met-Enk-Arg-Gly-Leu- and Met-Enk-Arg-Phe-like immunoreactivities. In the Bouin-fixed adrenal medulla, immunostaining of A cells with the 2 kinds of anti-Met-Enk-Arg-Gly-Leu sera (R-0171, NE2-206) was deeper than that of the NA cells. Immunostaining of A cells with the anti-Met-Enk-Arg-Phe serum (AP3-311) was apparently of the same intensity as that of NA cells. In 4% paraformaldehyde-fixed tissues, no correlation was seen between the distribution of E and NE cells and that of intensely and slightly Met-Enk-Arg-Gly-Leu and Met-Enk-Arg-Phe immunopositive cells. The masking of immunoreactive residues in chromaffin granules by an aldehyde-NE complex may be 1 of the major causes of the decreased opioid peptide-like immunoreactivity in the 4% paraformaldehyde-fixed adrenal medulla.