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NOSIP and Its Interacting Protein, eNOS, in the Rat Trachea and Lung

Peter König, Jürgen Dedio, Stefanie Oess, Tamara Papadakis, Axel Fischer, Werner Müller-Esterl and Wolfgang Kummer

Institut für Anatomie und Zellbiologie, Justus-Liebig-Universität, Giessen, Germany (PK,TP,WK); Disease Group Cardiovascular Diseases, Aventis Pharma GmbH, Frankfurt, Germany (JD); Institute for Biochemistry II, Johann Wolfgang Goethe University, Frankfurt, Germany (SO,WM-E); and Klinische Forschergruppe Allergologie, Charité, Berlin, Germany (AF)

Correspondence to: Peter König, Institut für Anatomie und Zellbiologie, Justus-Liebig-Universität Giessen, D-35385 Giessen, Germany. E-mail: peter.koenig{at}anatomie.med.uni-giessen.de

Endothelial nitric oxide synthase (eNOS), the major nitric oxide (NO)-generating enzyme of the vasculature, is regulated through multiple interactions with proteins, including caveolin-1, Hsp90, Ca²⁺-calmodulin, and the recently discovered eNOS-interacting protein, NOSIP. Previous studies indicate that NOSIP may contribute to the intricate regulation of eNOS activity and availability. Because eNOS has been shown to be abundantly expressed in the airways, we determined the expression and cellular localization of NOSIP in rat trachea and lung by RT-PCR and immunohistochemistry and examined the interaction of NOSIP with eNOS in lung by coimmunoprecipitation. In tracheal epithelium and lung, NOSIP mRNA expression was prevalent, as shown by RT-PCR, and the corresponding protein interacted with eNOS, as demonstrated by coimmunoprecipitation. Using immunohistochemistry, we found both NOSIP and eNOS immunoreactivity in ciliated epithelial cells of trachea and bronchi, while Clara cells showed immunoreactivity for NOSIP only. NOSIP and eNOS were present in vascular and bronchial smooth muscle cells of large arteries and airways, whereas endothelial cells, as well as bronchiolar and arteriolar smooth muscle cells, exclusively stained for NOSIP. Our results point to functional role(s) of NOSIP in the control of airway and vascular diameter, mucosal secretion, NO synthesis in ciliated epithelium, and, therefore, of mucociliary and bronchial function. (J Histochem Cytochem 53:155–164, 2005)

Key Words: NOSIP • ENOS-interacting protein • eNOS • lung • Clara cells • ciliated cells • smooth muscle cells

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