Immunohistochemical Evidence for the NO cGMP Signaling Pathway In Respiratory Ciliated Epithelia of Rat

Zhan, Xinhua; Li, Dechun; Johns, Roger A.

doi:10.1177/002215549904701103

Cited by 31 publications

(20 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although PKG-1 staining was additionally present in bronchial smooth muscle and alveoli, there was no detectable difference in staining intensity between control and CH lung tissue. Zhan et al (47) have similarly localized PKG-1␣ to pulmonary vascular and bronchial smooth muscle. However, in contrast to our present findings, PKG-1␤ was also found to be expressed in ciliated airway epithelial cells (47).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Pulmonary PKG-1 is upregulated following chronic hypoxia

Jernigan

Walker

Resta

2003

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

. Pulmonary PKG-1 is upregulated following chronic hypoxia. Am J Physiol Lung Cell Mol Physiol 285: L634-L642, 2003. First published May 23, 2003 10.1152/ ajplung.00328.2002.-Recent studies from our laboratory indicate that pulmonary vasodilatory responses to exogenous nitric oxide (NO) are attenuated following chronic hypoxia (CH) and that this NO-dependent vasodilation is mediated by cGMP. Similarly, we have demonstrated that CH attenuates vasodilatory responses to the cGMP analog 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP). We hypothesized that attenuated pulmonary vasodilation to 8-BrcGMP following CH is mediated by decreased protein kinase G-1 (PKG-1) expression/activity. Therefore, we examined vasodilatory responses to 8-BrcGMP (1 M) in isolated, saline-perfused lungs from control and CH (4 wk at barometric pressure of 380 mmHg) rats in the presence of the competitive PKG inhibitor Rp-␤-phenyl-1, N2-etheno-8-bromoguanosine 3',5'-cyclic monophosphorothionate (30 M) or the highly specific PKG inhibitor KT-5823 (10 M). PKG-1 expression and activity were determined in whole lung homogenates from each group, and vascular PKG-1 levels were assessed by quantitative immunohistochemistry. PKG inhibition with either Rp-8-Br-PET-cGMPS or KT-5823 diminished vasodilatory responses to 8-BrcGMP in lungs from both control and CH rats, thus indicating a role for PKG in mediating reactivity to 8-BrcGMP in each group. However, in contrast to our hypothesis, PKG-1 levels were approximately twofold greater in lungs from CH rats vs. controls, and furthermore, this upregulation was localized to the vasculature. This correlates with an increase in PKG activity following CH. We conclude that PKG-1 is involved in 8-BrcGMPmediated vasodilation; however, attenuated pulmonary vasodilation following CH is not associated with decreased expression/activity of PKG-1. isolated rat lung; nitric oxide; pulmonary hypertension; 8-bromoguanosine 3',5'-cyclic monophosphate; KT-5823; Rp-␤-phenyl-1, N2-etheno-8-bromoguanosine 3',5'-cyclic monophosphorothioate CHRONIC EXPOSURE TO HYPOXIA has been shown to augment pulmonary vasodilatory responses to endothelium-derived nitric oxide (EDNO)-dependent vasodilators (18, 26, 28, 32-34, 36, 37). EDNO is synthesized in the vasculature by endothelial nitric oxide synthase (eNOS), and several studies have demonstrated that chronic hypoxia (CH) is associated with increased pulmonary eNOS levels, gene expression, and activity (18,21,26,33,34,39,46). Consistent with elevated eNOS levels, nitric oxide (NO) synthesis appears to be greater in lungs isolated from CH rats compared with controls (18,26,44). In contrast to enhanced reactivity to our laboratory (19,34) and others (35) have demonstrated impaired responsiveness to exogenous NO following CH. However, the mechanism(s) responsible for this diminished reactivity is not fully understood.NO leads to pulmonary vascular smooth muscle (VSM) relaxation by stimulating soluble guanylyl cyclase (sGC) (7). The subsequent elevation in cGMP activates prot...

show abstract

Section: Discussionmentioning

confidence: 99%

“…Zhan et al (47) have similarly localized PKG-1␣ to pulmonary vascular and bronchial smooth muscle. However, in contrast to our present findings, PKG-1␤ was also found to be expressed in ciliated airway epithelial cells (47). The reason for this discrepancy is not clear but may be due to differences in protocols or antibodies used.…”

Section: Discussionmentioning

confidence: 99%

Pulmonary PKG-1 is upregulated following chronic hypoxia

Jernigan

Walker

Resta

2003

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

show abstract

“…PKG and its role in flagellar or ciliary beating have not been as widely studied as the other protein kinases; only PKG-Ib has been detected on rat airway epithelial cells (Zhan et al 1999). It has been reported that NO regulates CBF via activation of PKG (Li et al 2000) and that NO is essential for maintaining ciliary motility Zhan et al 2003).…”

Section: Discussionmentioning

confidence: 99%

Nitric Oxide-dependent Cilia Regulatory Enzyme Localization in Bovine Bronchial Epithelial Cells

Stout

Wyatt

Adams

et al. 2007

J Histochem Cytochem.

View full text Add to dashboard Cite

(JJA) S U M M A R Y 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 b (GC-b) 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-b, 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. (J Histochem Cytochem 55:433-442, 2007)

show abstract

“…Hsp90 is necessary for eNOS translocation to the apical membrane in response to increased luminal flow. Given the importance of eNOS-derived NO in the regulation of THAL NaCl absorption (35,38) and the role of eNOS in various physiological processes in epithelial cells of the respiratory tract, brain, and testis (14,29,55,57,60), this mechanism is likely to play an essential role in the regulation of NO levels in these cells.…”

Section: Discussionmentioning

confidence: 99%

“…We conclude that luminal flow induces eNOS translocation and activation in the THAL via PI3-kinase and that Hsp90 is involved in eNOS translocation to the apical membrane. nitric oxide; renal tubules; trafficking; phosphatidylinositol; heat shock protein; endothelial nitric oxide synthase IN POLARIZED EPITHELIAL CELLS of the kidney, respiratory tract, and testis, nitric oxide (NO) produced by endothelial NO synthase (eNOS) regulates important cellular functions (14,27,29,34,55,57,60). Thus eNOS activity and NO production must be tightly regulated in these cells.…”

mentioning

confidence: 99%

Luminal flow induces eNOS activation and translocation in the rat thick ascending limb. II. Role of PI3-kinase and Hsp90

Ortíz¹,

Hong²,

Garvin³

2004

American Journal of Physiology-Renal Physiology

View full text Add to dashboard Cite

Endothelial nitric oxide synthase (eNOS) regulates NaCl absorption by the thick ascending limb of the loop of Henle (THAL). We found that augmenting luminal flow induces eNOS activation and translocation to the apical membrane of THALs (Ortiz PA, Hong NJ, and Garvin JL. Am J Physiol Renal Physiol 287: F274 -F280, 2004). In other cells, eNOS activation by shear stress is mediated by phosphatidylinositol 3-OH kinase (PI3)-kinase. We hypothesized that luminal flow induces eNOS activation via PI3-kinase. Pretreatment of THALs with wortmannin, a PI3-kinase inhibitor, significantly reduced flow-induced nitric oxide (NO) release by 75% (from 53.6 Ϯ 6 to 13.2 Ϯ 5.7 pA/mm). Increasing luminal flow from 0 to 20 nl/min induced eNOS translocation to the apical membrane, whereas in the presence of wortmannin eNOS translocation was prevented (basolateral ϭ 32 Ϯ 2%, middle ϭ 38 Ϯ 1%, apical ϭ 30 Ϯ 1%, n ϭ 5, not significant vs. no flow). We next studied which PI3-kinase product mediates eNOS translocation. Addition of PI(3,4,5)P 3 (5 M) in the absence of flow increased NO levels (P Ͻ 0.05) and induced eNOS translocation to the apical membrane (from 40 Ϯ 4 to 60 Ϯ 2% of total eNOS, n ϭ 6, P Ͻ 0.05). Incubation with PI(3,4)P 2 or PI(4,5)P2 did not change eNOS localization. We next tested whether heat shock protein (Hsp)90 is involved in eNOS translocation. The Hsp90 inhibitor geldanamycin blocked flow-induced eNOS translocation to the apical membrane (n ϭ 6). Flow also induced translocation of Hsp90 to the apical membrane (from 35 Ϯ 2 to 57 Ϯ 2%; P Ͻ 0.05) in a PI3-kinasedependent manner. We conclude that luminal flow induces eNOS translocation and activation in the THAL via PI3-kinase and that Hsp90 is involved in eNOS translocation to the apical membrane. nitric oxide; renal tubules; trafficking; phosphatidylinositol; heat shock protein; endothelial nitric oxide synthase IN POLARIZED EPITHELIAL CELLS of the kidney, respiratory tract, and testis, nitric oxide (NO) produced by endothelial NO synthase (eNOS) regulates important cellular functions (14,27,29,34,55,57,60). Thus eNOS activity and NO production must be tightly regulated in these cells. We previously reported that NO acts as an autacoid to inhibit NaCl and bicarbonate absorption by the thick ascending limb of the loop of Henle (THAL) (31,32,39). More recently, we identified eNOS as the NOS isoform responsible for the regulation of NaCl absorption by the THAL (35, 38). We recently observed that increasing luminal flow acutely stimulated eNOS activity and induced translocation of eNOS from the basolateral membrane and cytoplasm to the apical membrane of isolated THALs (34a). However, the signaling cascade that mediates eNOS activation and translocation by flow is unknown.Regulation of eNOS has been studied in depth in vascular endothelial cells, where one of the most potent activators of eNOS is flow-induced shear stress (5, 46). The mechanism by which flow activates eNOS in these cells is independent of changes in intracellular calcium and involves activation of the pho...

show abstract

Immunohistochemical Evidence for the NO cGMP Signaling Pathway In Respiratory Ciliated Epithelia of Rat

Cited by 31 publications

References 26 publications

Pulmonary PKG-1 is upregulated following chronic hypoxia

Pulmonary PKG-1 is upregulated following chronic hypoxia

Nitric Oxide-dependent Cilia Regulatory Enzyme Localization in Bovine Bronchial Epithelial Cells

Luminal flow induces eNOS activation and translocation in the rat thick ascending limb. II. Role of PI3-kinase and Hsp90

Contact Info

Product

Resources

About