Localization of endothelial NOS at the basal microtubule membrane in ciliated epithelium of rat lung

Chun Xue, Suzanne J. Botkin, Roger A Johns

Research output: Contribution to journalArticle

Abstract

Nitric oxide (NO), an important cell messenger molecule, is formed endogenously in the lung airway. Three individual genes of NO synthase (NOS), which represent brain NOS (bNOS), inducible NOS (iNOS), and endothelial NOS (eNOS), have been reported in the cultured lung epithelium. Although studies in vivo showed that bNOS and iNOS were expressed and localized in the cytoplasm of bronchial epithelium, the expression and localization of eNOS remains to be determined. Therefore, we employed an eNOS monoclonal antibody whose immunospecificity was tested by both Western blot and preadsorption immunohistochemistry to immunostain rat lungs from fetus to adult. The results showed that eNOS immunoreactivity began to appear in the lung epithelium within 2 hr after birth. Six hours later (8 hr after birth), the NOS immunoreaction was concentrated near the surface of the ciliated epithelial cells. This staining pattern appeared in lungs at Day 1, Week 1, Week 2, and in adult rats. By electron microscopy, eNOS immunoreactivity was confirmed within ciliated epithelium and was shown to be associated with the basal microtubule membrane of the cilia. Nonciliated cells were not stained. Type H epithelial cells also contain eNOS immunoreactivity, which is primarily associated with rough endoplasmic reticulum and free ribosomes. However, macrophages in the lungs lacked eNOS immunoreactivity. This study demonstrated that eNOS was postnatally expressed in rat bronchial ciliated epithelium. The localization of eNOS at the basal membrane of ciliary microtubules suggests that eNOS my be involved in the function of epithelial cilia, consistent with previous physiological studies.

Original languageEnglish (US)
Pages (from-to)463-471
Number of pages9
JournalJournal of Histochemistry and Cytochemistry
Volume44
Issue number5
StatePublished - May 1996
Externally publishedYes

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Nitric Oxide Synthase
Microtubules
Epithelium
Lung
Membranes
Cilia
Epithelial Cells
Parturition
Rough Endoplasmic Reticulum
Brain
Ribosomes
Electron Microscopy
Nitric Oxide
Cytoplasm
Fetus
Western Blotting
Immunohistochemistry
Macrophages
Monoclonal Antibodies
Staining and Labeling

Keywords

  • Bronchial epithelium
  • Cilia
  • Endothelial NOS
  • Endothelium
  • Immunohistochemistry
  • Lung development
  • Macrophage
  • Type II alveolar epithelium

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Localization of endothelial NOS at the basal microtubule membrane in ciliated epithelium of rat lung. / Xue, Chun; Botkin, Suzanne J.; Johns, Roger A.

In: Journal of Histochemistry and Cytochemistry, Vol. 44, No. 5, 05.1996, p. 463-471.

Research output: Contribution to journalArticle

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N2 - Nitric oxide (NO), an important cell messenger molecule, is formed endogenously in the lung airway. Three individual genes of NO synthase (NOS), which represent brain NOS (bNOS), inducible NOS (iNOS), and endothelial NOS (eNOS), have been reported in the cultured lung epithelium. Although studies in vivo showed that bNOS and iNOS were expressed and localized in the cytoplasm of bronchial epithelium, the expression and localization of eNOS remains to be determined. Therefore, we employed an eNOS monoclonal antibody whose immunospecificity was tested by both Western blot and preadsorption immunohistochemistry to immunostain rat lungs from fetus to adult. The results showed that eNOS immunoreactivity began to appear in the lung epithelium within 2 hr after birth. Six hours later (8 hr after birth), the NOS immunoreaction was concentrated near the surface of the ciliated epithelial cells. This staining pattern appeared in lungs at Day 1, Week 1, Week 2, and in adult rats. By electron microscopy, eNOS immunoreactivity was confirmed within ciliated epithelium and was shown to be associated with the basal microtubule membrane of the cilia. Nonciliated cells were not stained. Type H epithelial cells also contain eNOS immunoreactivity, which is primarily associated with rough endoplasmic reticulum and free ribosomes. However, macrophages in the lungs lacked eNOS immunoreactivity. This study demonstrated that eNOS was postnatally expressed in rat bronchial ciliated epithelium. The localization of eNOS at the basal membrane of ciliary microtubules suggests that eNOS my be involved in the function of epithelial cilia, consistent with previous physiological studies.

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