Phosphorylation of Ser-279/282 and Tyr-265 positions on Cx43 as possible mediators of VEGF-165 inhibition of pregnancy-adapted Ca2+ burst function in ovine uterine artery endothelial cells

Boeldt, Derek S.; Grummer, Mary A.; Feng, Yi; Magness, Ronald R.; Bird, Ian M.

doi:10.1016/j.mce.2015.05.030

Cited by 20 publications

(26 citation statements)

References 30 publications

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“…Moreover, it is known that GJ assembly and disassembly are events highly regulated by a sequence of protein kinase activation and phosphorylation events. This kind of regulation, as extensively reported for Cx43, has a net effect of reducing GJ communication [102][103][104]. Therefore, we may also postulate that VEGF-A or other factors released by PRP may reduce GJ/Cx43 functionality affecting such phosphorylation events.…”

Section: Factors Released By Prp Possibly Modulating CX Expression Ansupporting

confidence: 69%

Platelet-Rich Plasma Modulates Gap Junction Functionality and Connexin 43 and 26 Expression During TGF-β1–Induced Fibroblast to Myofibroblast Transition: Clues for Counteracting Fibrosis

Squecco

Chellini

Idrizaj

et al. 2020

Cells

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Skeletal muscle repair/regeneration may benefit by Platelet-Rich Plasma (PRP) treatment owing to PRP pro-myogenic and anti-fibrotic effects. However, PRP anti-fibrotic action remains controversial. Here, we extended our previous researches on the inhibitory effects of PRP on in vitro transforming growth factor (TGF)-β1-induced differentiation of fibroblasts into myofibroblasts, the effector cells of fibrosis, focusing on gap junction (GJ) intercellular communication. The myofibroblastic phenotype was evaluated by cell shape analysis, confocal fluorescence microscopy and Western blotting analyses of α-smooth muscle actin and type-1 collagen expression, and electrophysiological recordings of resting membrane potential, resistance, and capacitance. PRP negatively regulated myofibroblast differentiation by modifying all the assessed parameters. Notably, myofibroblast pairs showed an increase of voltage-dependent GJ functionality paralleled by connexin (Cx) 43 expression increase. TGF-β1-treated cells, when exposed to a GJ blocker, or silenced for Cx43 expression, failed to differentiate towards myofibroblasts. Although a minority, myofibroblast pairs also showed not-voltage-dependent GJ currents and coherently Cx26 expression. PRP abolished the TGF-β1-induced voltage-dependent GJ current appearance while preventing Cx43 increase and promoting Cx26 expression. This study adds insights into molecular and functional mechanisms regulating fibroblast-myofibroblast transition and supports the anti-fibrotic potential of PRP, demonstrating the ability of this product to hamper myofibroblast generation targeting GJs.

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Section: Factors Released By Prp Possibly Modulating CX Expression Ansupporting

confidence: 69%

Platelet-Rich Plasma Modulates Gap Junction Functionality and Connexin 43 and 26 Expression During TGF-β1–Induced Fibroblast to Myofibroblast Transition: Clues for Counteracting Fibrosis

Squecco

Chellini

Idrizaj

et al. 2020

Cells

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“…Common signaling ‘culprits’ that phosphorylate these inhibitory residues are Src, ERK, and PKC (Bird et al 2013). This has been corroborated in the ovine uterine artery endothelial cell model, where administration of VEGF leads to both phosphorylation of Cx43 and reduced sustained phase Ca 2+ signaling (Boeldt et al 2015). Inhibitors of Src and ERK signaling reversed phosphorylation of Cx43 at their respective target residues and also rescued Ca 2+ signaling to levels equivalent to control (Boeldt et al 2015).…”

Section: Effects Of Hormones Of Pe On the Endotheliummentioning

confidence: 75%

“…This has been corroborated in the ovine uterine artery endothelial cell model, where administration of VEGF leads to both phosphorylation of Cx43 and reduced sustained phase Ca 2+ signaling (Boeldt et al 2015). Inhibitors of Src and ERK signaling reversed phosphorylation of Cx43 at their respective target residues and also rescued Ca 2+ signaling to levels equivalent to control (Boeldt et al 2015). More detailed studies on other hormones (growth factors and cytokines) associated with of PE are warranted, to assess the universality of these initial observations, and whether they are indeed occurring in PE.…”

Section: Effects Of Hormones Of Pe On the Endotheliummentioning

confidence: 75%

Vascular adaptation in pregnancy and endothelial dysfunction in preeclampsia

Boeldt

Bird

2017

Journal of Endocrinology

249

152

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Maternal vascular adaptation to pregnancy is critically important in order to expand the capacity for blood flow through the uteroplacental unit to meet the needs of the developing fetus. Failure of the maternal vasculature to properly adapt can result in hypertensive disorders of pregnancy such as preeclampsia (PE). Herein, we review the endocrinology of maternal adaptation to pregnancy and contrast this with that of PE. Our focus is specifically on those hormones with directly impact endothelial cell function and dysfunction, as endothelial cell dysfunction is a hallmark of PE. A variety of growth factors and cytokines are present in normal vascular adaptation to pregnancy but have also been shown to be circulating at abnormal levels in PE pregnancies. Many of these factors promote endothelial dysfunction when present at abnormal levels by acutely inhibiting key Ca2+ signaling events and chronically promoting the breakdown of endothelial cell-cell contacts. Increasingly, our understanding of how the contributions of the placenta, immune cells, and the endothelium itself promote the endocrine milieu of PE is becoming more clear. We then describe in detail how the complex endocrine environment of PE effects endothelial cell function, why this has contributed to the difficulty in fully understanding and treating this disorder, and how a focus on signaling convergence points of many hormones may be a more successful treatment strategy.

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“…These phosphorylations cause inhibition of intercellular communication which is well described for tumor cells (52). Previous reports have shown that cSrc phosphorylation suppresses gap junctional communication between endothelial cells (54), and the importance of Y265 has been suggested (8). Furthermore, paired cardiomyocytes from hamster in late-stage congestive heart failure have constitutively activated cSrc but Cx43 tyrosine phosphorylation was associated with suppressed intercellular communication (57).…”

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confidence: 90%

Na-K-ATPase regulates intercellular communication in the vascular wall via cSrc kinase-dependent connexin43 phosphorylation

Hangaard

Bouzinova

Staehr

et al. 2017

American Journal of Physiology-Cell Physiology

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Communication between vascular smooth muscle cells (VSMCs) is dependent on gap junctions and is regulated by the Na-K-ATPase. The Na-K-ATPase is therefore important for synchronized VSMC oscillatory activity, i.e., vasomotion. The signaling between the Na-K-ATPase and gap junctions is unknown. We tested here the hypothesis that this signaling involves cSrc kinase. Intercellular communication was assessed by membrane capacitance measurements of electrically coupled VSMCs. Vasomotion in isometric myograph, input resistance, and synchronized [Ca] transients were used as readout for intercellular coupling in rat mesenteric small arteries in vitro. Phosphorylation of cSrc kinase and connexin43 (Cx43) were semiquantified by Western blotting. Micromole concentration of ouabain reduced the amplitude of norepinephrine-induced vasomotion and desynchronized Ca transients in VSMC in the arterial wall. Ouabain also increased input resistance in the arterial wall. These effects of ouabain were antagonized by inhibition of tyrosine phosphorylation with genistein, PP2, and by an inhibitor of the Na-K-ATPase-dependent cSrc activation, pNaKtide. Moreover, inhibition of cSrc phosphorylation increased vasomotion amplitude and decreased the resistance between cells in the vascular wall. Ouabain inhibited the electrical coupling between A7r5 cells, but pNaKtide restored the electrical coupling. Ouabain increased cSrc autophosphorylation of tyrosine 418 (Y418) required for full catalytic activity whereas pNaKtide antagonized it. This cSrc activation was associated with Cx43 phosphorylation of tyrosine 265 (Y265). Our findings demonstrate that Na-K-ATPase regulates intercellular communication in the vascular wall via cSrc-dependent Cx43 tyrosine phosphorylation.

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Phosphorylation of Ser-279/282 and Tyr-265 positions on Cx43 as possible mediators of VEGF-165 inhibition of pregnancy-adapted Ca2+ burst function in ovine uterine artery endothelial cells

Cited by 20 publications

References 30 publications

Platelet-Rich Plasma Modulates Gap Junction Functionality and Connexin 43 and 26 Expression During TGF-β1–Induced Fibroblast to Myofibroblast Transition: Clues for Counteracting Fibrosis

Platelet-Rich Plasma Modulates Gap Junction Functionality and Connexin 43 and 26 Expression During TGF-β1–Induced Fibroblast to Myofibroblast Transition: Clues for Counteracting Fibrosis

Vascular adaptation in pregnancy and endothelial dysfunction in preeclampsia

Na-K-ATPase regulates intercellular communication in the vascular wall via cSrc kinase-dependent connexin43 phosphorylation

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