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¹ Departments of Pharmacology and Toxicology
² Pathology, Otto-von-Guericke-University, Leipzieger Strasse 44, 39120 Magdeburg, Germany
³ Department of Medicine II, Klinikum rechts der Isar, Technische Universität München, 81675 München, Germany
⁴ Obstetrics and Gynecology, Otto-von-Guericke-University, Leipziger Strasse 44, 39120 Magdeburg, Germany
⁵ Department of Pathology, Charité – Universitätsmedizin Berlin, Schumannstrasse 20/21, 10117 Berlin, Germany

(Requests for offprints should be addressed to S Schulz; Email: stefan.schulz{at}medizin.uni-magdeburg.de)

The biological effects of neurotensin (NT) are mediated by two distinct G protein-coupled receptors, NTS₁ and NTS₂. Although it is well established that neurotensin inhibits gastric acid secretion in man, the plasma membrane receptor mediating these effects has not been visualized yet. We developed and characterized a novel antipeptide antibody to the carboxy-terminal region of the human NTS₂ receptor. The cellular and subcellular distribution of NTS₂ receptors was evaluated in various human gastrointestinal tissues. Specificity of the antiserum was demonstrated by (1) detection of a broadband migrating at M_r 90 000–100 000 in Western blots of membranes from NTS₂-expressing tissues; (2) cell-surface staining of NTS₂-transfected cells; (3) translocation of NTS₂ receptor immunostaining after agonist exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with its immunizing peptide. In the gastrointestinal tract, NTS₂ receptor immunoreactivity was highly abundant in parietal cells of the gastric mucosa, in neuroendocrine cells of the stomach small and large intestine, and in cells of the exocrine pancreas. NTS₂ receptors were clearly located in the plasma membrane and uniformly present on nearly all target cells. The presence of NTS₂ receptors was rarely detected in human tumors. This is the first localization of NTS₂ receptors in human formalin-fixed, paraffin-embedded tissues at the cellular level. The abundant expression of low-affinity NTS₂ receptors on the plasma membrane of human parietal cells provides a morphological substrate for the direct inhibition of gastric acid secretion observed after i.v. administration of neurotensin.