VEGF Induces Nuclear Translocation of Flk-1/KDR, Endothelial Nitric Oxide Synthase, and Caveolin-1 in Vascular Endothelial Cells

Feng, Yiyu; Venema, Virginia J.; Venema, Richard C.; Tsai, Nai Tse; Caldwell, Ruth B.

doi:10.1006/bbrc.1998.9790

Cited by 197 publications

(153 citation statements)

References 43 publications

Supporting

Mentioning

145

Contrasting

Unclassified

Order By: Relevance

“…A similar translocation could be observed in endothelial cells treated with VEGF for several hours (ref. 23 and N.R., unpublished results). Although the biological function of the nuclear VEGFR2 is yet to be defined, nuclear translocation of additional tyrosine kinase receptors such as fibroblast growth factor and epidermal growth factor was demonstrated also, a translocation that was accompanied by transcriptional activity of the receptors (24,25).…”

Section: Resultsmentioning

confidence: 66%

VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells

Shay-Salit

Shushy

Wolfovitz

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

299

221

View full text Add to dashboard Cite

Blood-flow interactions with the vascular endothelium represents a specialized example of mechanical regulation of cell function that has important physiological and pathophysiological cardiovascular consequences. Yet, the mechanisms of mechanostransduction are not understood fully. This study shows that shear stress induces a rapid induction as well as nuclear translocation of the vascular endothelial growth factor (VEGF) receptor 2 and promotes the binding of the VEGF receptor 2 and the adherens junction molecules, VE-cadherin and ␤-catenin, to the endothelial cytoskeleton. These changes are accompanied by the formation of a complex containing the VEGF receptor 2-VE-cadherin-␤-catenin. In endothelial cells lacking VE-cadherin, shear stress did not augment nuclear translocation of the VEGF receptor 2 and phosphorylation of Akt1 and P38 as well as transcriptional induction of a reporter gene regulated by a shear stress-responsive promoter. These results suggest that VEGF receptor 2 and the adherens junction act as shear-stress cotransducers, mediating the transduction of shearstress signals into vascular endothelial cells.

show abstract

Section: Resultsmentioning

confidence: 66%

VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells

Shay-Salit

Shushy

Wolfovitz

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

299

221

View full text Add to dashboard Cite

show abstract

“…In addition, these two agonists also induce rapid eNOS translocation from the plasma membrane to the cytoplasm of cultured endothelial cells (10,19). However, it was not known whether PI3-kinase was involved in eNOS translocation in these cells.…”

Section: Discussionmentioning

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

“…The immunohistochemical analysis showed that major as well as rare liver disorders examined in this study were associated with profound changes in the cellular distribution of eNOS, leading to its translocation to hepatocyte nuclei. Interestingly, growth factors such as vascular-endothelial growth factor are known to cause nuclear translocation of eNOS in vascular endothelium (25). The significance of this observation is as yet unclear.…”

Section: Discussionmentioning

confidence: 99%

Distribution of nitric oxide synthase in normal and cirrhotic human liver

McNaughton

Puttagunta

Martínez-Cuesta

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

149

View full text Add to dashboard Cite

Chronic liver disorders represent a serious health problem, considering that 300 million people worldwide are hepatitis B virus carriers, and 8,000 -10,000 patients per year, in the U.S. alone, die as a result of liver failure caused by hepatitis C infection. Nitric oxide synthase (NOS) regulates hepatic vasculature; however, the patterns of expression and activity of NOS proteins in healthy and diseased human livers are unknown. Sections of diseased (n ‫؍‬ 42) and control livers (n ‫؍‬ 14) were collected during orthotopic liver transplants and partial hepatectomy. The diseased sections included alcoholic cirrhosis, viral hepatitis, cholestasis, acute necrosis, and uncommon pathologies including ␣1-anti-trypsin disorder. The endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS) were studied by using the citrulline assay, Western immunoblot, immunohistochemistry, and in situ hybridization. The systemic generation of plasma NO metabolites was measured by HPLC. In control livers, Ca 2؉ -dependent and -independent NOS activities were identified by Western analysis as eNOS and iNOS, respectively. The eNOS was uniformly distributed in the hepatocytes and also detected in the endothelium of hepatic arteries, terminal hepatic venules, sinusoids, and in biliary epithelium. The iNOS was detected in hepatocytes and localized mainly in the periportal zone of the liver acinus. This pattern of distribution of eNOS and iNOS in normal liver was confirmed by in situ hybridization. In diseased livers, there was a significant increase in Ca 2؉ -independent NOS with the corresponding strong appearance of iNOS in the cirrhotic areas. The eNOS was translocated to hepatocyte nuclei. Thus, eNOS and iNOS proteins are differentially expressed in healthy human liver, and this expression is significantly altered in cirrhotic liver disorders.

show abstract

VEGF Induces Nuclear Translocation of Flk-1/KDR, Endothelial Nitric Oxide Synthase, and Caveolin-1 in Vascular Endothelial Cells

Cited by 197 publications

References 43 publications

VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells

VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells

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

Distribution of nitric oxide synthase in normal and cirrhotic human liver

Contact Info

Product

Resources

About