Production of antiserum to [D-Ala2]deltorphin I and its immunohistochemical application to the mouse brain.

Antiserum to haptenic [D-Ala2]deltorphin I (DADTI: Tyr-D-Ala-Phe-Asp-Val-Val-Gly.NH2), a highly selective ligand for delta opioid receptors, was produced in rabbits. By immunospot assay, the antiserum recognized 62.5 pmol DADTI but failed to react even with 4 nmol carrier protein of the immunogen. Although the antiserum reacted equally with an isomer [L-Ala2]deltorphin I, virtually no cross-reaction occurred with other analogues such as [D-Ala2]deltorphin II (Tyr-D-Ala-Phe-Glu-Val-Val-Gly.NH2) and similar peptides lacking the C-terminal glycine amide. Therefore, the major epitope for immunorecognition appeared to be in the C-terminal region which is known to be the specific domain for delta opioid receptor selectivity. Immunohistochemical study using this antiserum revealed positive neuronal structures in some specific systems of the mouse brain.