title: Immunocytochemical identification of low-affinity NTS2 neurotensin receptors in parietal cells of human gastric mucosa.
abstract: The biological effects of neurotensin (NT) are mediated by two distinct G protein-coupled receptors, NTS(1) and NTS(2). 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 NTS(2) receptor. The cellular and subcellular distribution of NTS(2) 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 NTS(2)-expressing tissues; (2) cell-surface staining of NTS(2)-transfected cells; (3) translocation of NTS(2) receptor immunostaining after agonist exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with its immunizing peptide. In the gastrointestinal tract, NTS(2) 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. NTS(2) receptors were clearly located in the plasma membrane and uniformly present on nearly all target cells. The presence of NTS(2) receptors was rarely detected in human tumors. This is the first localization of NTS(2) receptors in human formalin-fixed, paraffin-embedded tissues at the cellular level. The abundant expression of low-affinity NTS(2) 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.
authors.affiliation: Department of Pharmacology and Toxicology, Otto-von-Guericke-University, Magdeburg, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schulz
authors.forename: Christoph
authors.initials: C
authors.lastname: Röcken
authors.forename: Matthias P A
authors.initials: MP
authors.lastname: Ebert
authors.forename: Solveig
authors.initials: S
authors.lastname: Schulz
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002462:Cell Membrane / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007340:Insulinoma / Q000737:chemistry; D007422:Intestines / Q000737:chemistry; D008297:Male; D009496:Neurotensin / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016612:Paraffin Embedding; D010295:Parietal Cells, Gastric / Q000737:chemistry*; D011485:Protein Binding; D018028:Receptors, Neurotensin / Q000032:analysis* / Q000235:genetics; D013274:Stomach Neoplasms / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; C490985:NTSR2 protein, human; D018028:Receptors, Neurotensin; D009496:Neurotensin