Inhibition of Endothelial Cell Migration, Intercellular Communication, and Vascular Tube Formation by Thromboxane A2

Ashton, Anthony W.; Yokota, Ryoji; John, Gareth W.; Zhao, Shuai; Suadicani, Sylvia O.; Spray, David C.; Ware, J. Anthony

doi:10.1074/jbc.274.50.35562

Cited by 140 publications

(106 citation statements)

References 41 publications

(39 reference statements)

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“…TXA 2 receptor stimulation has been linked to alterations in the distribution of gap junctions in human endothelial cells, which might impair functional coupling (Ashton et al, 1999). This seems an unlikely explanation for the selective loss of SK Ca -evoked hyperpolarization, unless these channels are located on separate endothelial cells to the IK Ca channels, as IK Ca -linked effects persisted.…”

Section: Discussionmentioning

confidence: 99%

Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery

Crane

Garland

2004

British J Pharmacology

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1 The possibility that thromboxane (TXA 2 ) receptor stimulation causes differential block of the SK Ca and IK Ca channels which underlie EDHF-mediated vascular smooth muscle hyperpolarization and relaxation was investigated in the rat isolated mesenteric artery. 2 Acetylcholine (30 nM-3 mM ACh) or cyclopiazonic acid (10 mM CPA, SERCA inhibitor) were used to stimulate EDHF-evoked smooth muscle hyperpolarization. In each case, this led to maximal hyperpolarization of around 20 mV, which was sensitive to block with 50 nM apamin and abolished by repeated stimulation of mesenteric arteries with the thromboxane mimetic, U46619 (30 nM-0.1 mM), but not the a 1 -adrenoceptor agonist phenylephrine (PE). 3 The ability of U46619 to abolish EDHF-evoked smooth muscle hyperpolarization was prevented by prior exposure of mesenteric arteries to the TXA 2 receptor antagonist 1 mM SQ29548. 4 Similar-sized smooth muscle hyperpolarization evoked with the SK Ca activator 100 mM riluzole was also abolished by prior stimulation with U46619, while direct muscle hyperpolarization in response to either levcromakalim (1 mM, K ATP activator) or NS1619 (40 mM, BK Ca activator) was unaffected. 5 During smooth muscle contraction and depolarization to either PE or U46619, ACh evoked concentration-dependent hyperpolarization (to À67 mV) and complete relaxation. These responses were well maintained during repeated stimulation with PE, but with U46619 there was a progressive decline, so that during a third exposure to U46619 maximum hyperpolarization only reached -52 mV and relaxation was reduced by 20%. This relaxation could now be blocked with charybdotoxin alone. The latter responses could be mimicked with 300 mM 1-EBIO (IK Ca activator), an action not modified by exposure to U46619. 6 An early consequence of TXA 2 receptor stimulation is a reduction in the arterial hyperpolarization and relaxation attributed to EDHF. This effect appears to reflect a loss of SK Ca activity.

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Section: Discussionmentioning

confidence: 99%

Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery

Crane

Garland

2004

British J Pharmacology

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“…Human umbilical vein endothelial cells (HUVECs) were isolated from the human umbilical vein and cultured as described previously (24,26). The cells were grown in MCDB131 supplemented with 5% heat-inactivated human serum, 20% heat-inactivated newborn calf serum, 150 g/ml endothelial cell growth supplement, 5 units/ml heparin sodium, 100 units/ml penicillin, and 100 g/ml streptomycin.…”

Section: Methodsmentioning

confidence: 99%

Identification of a Novel Domain at the N Terminus of Caveolin-1 That Controls Rear Polarization of the Protein and Caveolae Formation

Sun¹,

Flynn

Castranova

et al. 2007

Journal of Biological Chemistry

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When cells are migrating, caveolin-1, the principal protein component of caveolae, is excluded from the leading edge and polarized at the cell rear. The dynamic feature depends on a specific sequence motif that directs intracellular trafficking of the protein. Deletion mutation analysis revealed a putative polarization domain at the N terminus of caveolin-1, between amino acids 32-60. Alanine substitution identified a minimal sequence of 10 residues ( 46 TKEIDLVNRD 55 ) necessary for caveolin-1 rear polarization. Interestingly, deletion of amino acids 1-60 did not prevent the polarization of caveolin-1 in human umbilical vein endothelial cells or wild-type mouse embryonic fibroblasts because of an interaction of Cav 61-178 mutant with endogenous caveolin-1. Surprisingly, expression of the depolarization mutant in caveolin-1 null cells dramatically impeded caveolae formation. Furthermore, knockdown of caveolae formation by methyl-␤-cyclodextrin failed to prevent wild-type caveolin-1 rear polarization. Importantly, genetic depletion of caveolin-1 led to disoriented migration, which can be rescued by full-length caveolin-1 but not the depolarization mutant, indicating a role of caveolin-1 polarity in chemotaxis. Thus, we have identified a sequence motif that is essential for caveolin-1 rear polarization and caveolae formation.Caveolae are specific microdomains of the plasma membrane that were discovered more than 50 years ago (1). In endothelial cells, numerous vesicles appeared to derive from the uniformly flask-shaped invaginations, suggesting the endocytic potential of caveolae (2). Although the function of caveolae as transport vesicles mediating endocytosis and transcytosis remained obscure (3, 4), the identification, cloning, and characterization of caveolar coat proteins, caveolins, has increased our knowledge of caveolae, and a good body of evidence implicates caveolae in a specialized form of delivery of membrane components, extracellular ligands, bacterial toxins, and nonenveloped virus in several cell types (5-8). The caveolae-mediated endocytic pathway differs from that mediated by clathrincoated pits. It is sensitive to protein kinase C inhibitors and cholesterol depletion (by filipin), and in some cells it is involved in the activation of protein tyrosine kinases (9). Phosphorylation at tyrosine 14 of caveolin-1 (Cav-1) 2 may be required for the internalization of caveolae. The mechanism controlling caveolae trafficking remains unclear, but it apparently involves both microtubule and actin cytoskeletons (10). Surprisingly, using Cav-1 as a marker for caveolae, recent studies demonstrate that caveolae are rich in a variety of signaling molecules, with the implication that caveolae may function in the regulation of signal transduction. Given these views, an attractive hypothesis would be whether caveolae could carry signaling machinery to different locations of the cell to spatially organize signaling events. Indeed, Anderson and colleagues (11) have shown recently that concomitant with the relocation o...

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“…In addition, 20-min treatment of the CA rings isolated from control mice with 18␤-glycyrrhetinic acid (18␤-GA, 50 M), a nonspecific gap junction inhibitor (1,46), markedly attenuated ACh-induced vasodilatation (Fig. 4).…”

Section: Decreased Cx37 and Cx40 Protein Expression In Coronarymentioning

confidence: 99%

Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice

Platoshyn

Suárez

Yuan

et al. 2008

American Journal of Physiology-Cell Physiology

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Dillmann WH. Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice. Am J Physiol Cell Physiol 295: C221-C230, 2008. First published May 7, 2008 doi:10.1152/ajpcell.00433.2007.-Vascular endothelial cells (ECs) play a major role in regulating vascular tone and in revascularization. There is increasing evidence showing endothelial dysfunction in diabetes, although little is known about the contribution of connexins (Cxs) to vascular complications in the diabetic heart. This study was designed to investigate the role of Cxs in coronary endothelial dysfunction in diabetic mice. Coronary ECs isolated from diabetic mice exhibit lowered protein levels of Cx37 and Cx40 (but not Cx43) and a loss of gap junction intercellular communication (GJIC). Vasodilatation induced by the assumed contribution of ECdependent hyperpolarization was significantly reduced in the diabetic coronary artery (CA). Cx40-specific inhibitory peptide 40 GAP27 strongly attenuated endothelium-dependent relaxation in diabetic CA at the concentration that does not affect the relaxation in control CA, suggesting that the total amount of Cx40 is lower in diabetic CA than in control CA. In diabetic mice, coronary capillary density was significantly decreased in vivo. In vitro, GJIC inhibitor attenuated the ability of EC capillary network formation. High-glucose treatment caused a decrease in Cx40 protein expression in ECs and impaired endothelial capillary network formation, which was restored by Cx40 overexpression. Furthermore, we found that the hyperglycemia-induced decrease in Cx40 was associated with inhibited protein expression of Sp1, a transcriptional factor that regulates Cx40 expression. These data suggest that downregulation of Cx40 protein expression and resultant inhibition of GJIC contribute to coronary vascular dysfunction in diabetes.

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Inhibition of Endothelial Cell Migration, Intercellular Communication, and Vascular Tube Formation by Thromboxane A2

Cited by 140 publications

References 41 publications

Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery

Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery

Identification of a Novel Domain at the N Terminus of Caveolin-1 That Controls Rear Polarization of the Protein and Caveolae Formation

Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice

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Inhibition of Endothelial Cell Migration, Intercellular Communication, and Vascular Tube Formation by Thromboxane A2

Cited by 140 publications

References 41 publications

Thromboxane receptor stimulation associated with loss of SKCa activity and reduced EDHF responses in the rat isolated mesenteric artery

Thromboxane receptor stimulation associated with loss of SKCa activity and reduced EDHF responses in the rat isolated mesenteric artery

Identification of a Novel Domain at the N Terminus of Caveolin-1 That Controls Rear Polarization of the Protein and Caveolae Formation

Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice

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Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery

Thromboxane receptor stimulation associated with loss of SK_Ca activity and reduced EDHF responses in the rat isolated mesenteric artery