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JHC exPRESS: First Published May 1, 2006. doi:10.1369/jhc.6A6966.2006
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A more recent version of this article appeared on September 1, 2006.
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Immunolocalization of Full-length NK1 Tachykinin Receptors in Human Tumors

Stefan Schulz 1*, Ralf Stumm 1, Christoph Röcken 1, Christian Mawrin 1 and Solveig Schulz 1

1 Departments of Pharmacology and Toxicology (StS,RS), Pathology (CR), Neuropathology (CM), and Obstetrics and Gynecology (SoS), Otto-von-Guericke-University, Magdeburg, Germany

* To whom correspondence should be addressed. E-mail: Stefan.Schulz{at}Medizin.Uni-Magdeburg.de.

Submitted on March 12, 2006
Accepted on 19 April 2006


  Abstract
The biological effects of substance P (SP) are mediated by the neurokinin-1 (NK1) receptor which exists as full-length and as 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 Mr 70,000-90,000 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 preadsorbtion of the antibody with its immunizing peptide. The distribution of NK1 receptors was investigated in seventy-two formalin-fixed, paraffin-embedded human tumors showing that NK1 receptors were frequently expressed in glioblastomas, breast and pancreatic carcinomas. Immunoreactive NK1 receptors were clearly confined to the plasma membrane and uniformly present on nearly all tumor cells. The 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.

Key Words: substance P, tachykinin, NK1 receptor, neoplasm, antibody, immunocytochemistry, Western blot


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[Abstract] [Full Text] [PDF]



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