Nitric oxide-dependent cilia regulatory enzyme localization in bovine bronchial epithelial cells

Sarah L. Stout, Todd A Wyatt, Jennifer J. Adams, Joseph Harold Sisson

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Airway epithelial-derived nitric oxide (NO), through the activation of nucleotide cyclases and downstream kinases, stimulates ciliary beating, yet the precise locations of these enzymes are unknown. We hypothesized that these NO-activated enzymes are located within, or adjacent to, the ciliary axoneme. Immunohistochemistry of intact ciliated cells revealed that endothelial-type nitric oxide synthase (eNOS), the RII isoform of the cAMP-dependent protein kinase (PKA-RII), the type I isoform of the cGMP-dependent protein kinase (PKG-I), and guanylate cyclase β (GC-β) all colocalized with pericentrin to the basal body. In contrast, the PKA-RI isoform and the PKG-II isoform localized to ciliary axonemes. Western blot analysis of isolated demembranated ciliary preparations detected eNOS, GC-β, and both isoforms of PKA and PKG. An A-kinase-anchoring protein was also detected. Our findings suggest that these enzymes are sequestered close to their points of action into a discrete ciliary metabolon, enabling targeted phosphorylation and efficient upregulation of ciliary beating.

Original languageEnglish (US)
Pages (from-to)433-442
Number of pages10
JournalJournal of Histochemistry and Cytochemistry
Volume55
Issue number5
DOIs
StatePublished - May 1 2007

Fingerprint

Cilia
Nitric Oxide
Protein Isoforms
Epithelial Cells
Axoneme
Enzymes
Nitric Oxide Synthase Type III
Guanylate Cyclase
Cyclic GMP-Dependent Protein Kinase Type I
Basal Bodies
Cyclic AMP-Dependent Protein Kinases
Protein Kinases
Phosphotransferases
Up-Regulation
Nucleotides
Western Blotting
Immunohistochemistry
Phosphorylation

Keywords

  • Alcohol
  • Cilia
  • Nitric oxide
  • cAMP-dependent protein kinase
  • cGMP-dependent protein kinase

ASJC Scopus subject areas

  • Anatomy
  • Histology

Cite this

Nitric oxide-dependent cilia regulatory enzyme localization in bovine bronchial epithelial cells. / Stout, Sarah L.; Wyatt, Todd A; Adams, Jennifer J.; Sisson, Joseph Harold.

In: Journal of Histochemistry and Cytochemistry, Vol. 55, No. 5, 01.05.2007, p. 433-442.

Research output: Contribution to journalArticle

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