Na-K-ATPase regulates intercellular communication in the vascular wall via CSRC Kinase-Dependent connexin43 phosphorylation
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Na-K-ATPase regulates intercellular communication in the vascular wall via CSRC Kinase-Dependent connexin43 phosphorylation. / Hangaard, Lise; Bouzinova, Elena V.; Staehr, Christian; Dam, Vibeke S.; Kim, Sukhan; Xie, Zijian; Aalkjaer, Christian; Matchkov, Vladimir V.
In: American Journal of Physiology - Cell Physiology, Vol. 312, No. 4, 04.2017, p. C385-C397.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Na-K-ATPase regulates intercellular communication in the vascular wall via CSRC Kinase-Dependent connexin43 phosphorylation
AU - Hangaard, Lise
AU - Bouzinova, Elena V.
AU - Staehr, Christian
AU - Dam, Vibeke S.
AU - Kim, Sukhan
AU - Xie, Zijian
AU - Aalkjaer, Christian
AU - Matchkov, Vladimir V.
PY - 2017/4
Y1 - 2017/4
N2 - 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 [Ca2+]i 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 Ca2+ 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.
AB - 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 [Ca2+]i 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 Ca2+ 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.
KW - Artery
KW - CSrc kinase signaling
KW - Gap junctions
KW - Na-K-ATPase
KW - Vasomotion
U2 - 10.1152/ajpcell.00347.2016
DO - 10.1152/ajpcell.00347.2016
M3 - Journal article
C2 - 28122732
AN - SCOPUS:85017287451
VL - 312
SP - C385-C397
JO - American Journal of Physiology: Cell Physiology
JF - American Journal of Physiology: Cell Physiology
SN - 0363-6143
IS - 4
ER -
ID: 195965707