Immunolocalization of full-length NK1 tachykinin receptors in human tumors.
Biological effects of substance P (SP) are mediated by the neurokinin-1 (NK1) receptor that exists as a full-length and as a carboxy-terminally truncated isoform in humans. Although NK1 receptor mRNA and binding sites have been detected in certain malignancies, little is known about the cellular and subcellular localization of NK1 receptor protein in human neoplastic tissues. We developed and characterized a novel anti-peptide antibody to the carboxy-terminal region of the human full-length NK1 receptor. Specificity of the antiserum was demonstrated by (1) detection of a broad band migrating at molecular mass 70,000-90,000 Da in Western blots of membranes from NK1-expressing tissues; (2) cell-surface staining of NK1-transfected cells; (3) translocation of NK1 receptor immunostaining after SP exposure; and (4) abolition of tissue immunostaining by preadsorption of the antibody with its immunizing peptide. Distribution of NK1 receptors was investigated in 72 formalin-fixed, paraffin-embedded human tumors showing that NK1 receptors were frequently expressed in glioblastomas and breast and pancreatic carcinomas. Immunoreactive NK1 receptors were clearly confined to the plasma membrane and uniformly present on nearly all tumor cells. Development of this novel NK1 receptor antibody allows the efficient localization of NK1 receptor protein in human formalin-fixed, paraffin-embedded tissues. NK1 receptor visualization with this simple and rapid immunohistochemical method will facilitate identification of tumors with a sufficient receptor overexpression for diagnostic or therapeutic intervention using SP analogs.
Schulz
Stefan
S
Department of Pharmacology and Toxicology, Otto-von-Guericke-University, Leipziger Strasse 44, 39120 Magdeburg, Germany. Stefan.Schulz@Medizin.Uni-Magdeburg.de
Stumm
Ralf
R
Röcken
Christoph
C
Mawrin
Christian
C
Schulz
Solveig
S
D000906:Antibodies; D000918:Antibody Specificity; D002460:Cell Line; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D009369:Neoplasms / Q000378:metabolism*; D016612:Paraffin Embedding; D010446:Peptide Fragments / Q000276:immunology; D018040:Receptors, Neurokinin-1 / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D005404:Fixatives; D010446:Peptide Fragments; D018040:Receptors, Neurokinin-1; D005557:Formaldehyde