Src Regulates Distinct Pathways for Cell Volume Control through Vav andPhospholipaseCγ

Barfod, Elisabeth T.; Moore, Ann L.; Melnick, Richard F.; Lidofsky, Steven D.

doi:10.1074/jbc.m411717200

Cited by 24 publications

(35 citation statements)

References 59 publications

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“…8). Nevertheless, changes in the structure of the F-actin cytoskeleton are a prerequisite for swelling-induced ion channel activity (1,7,8,13), probably through a process of vesicle-mediated insertion and retrieval of proteins (7). This process is enhanced by PMA (11) and impaired by mutation of PKC activity (16), implying that PKC is a critical regulatory element for efficient cytoskeleton-mediated membrane protein sorting.…”

Section: Discussionmentioning

confidence: 99%

“…Neither can we exclude the possibility that product(s) of phospholipase C (PLC)-mediated PIP 2 hydrolysis, such as diacylglycerol or inositol 1,4,5-trisphosphate exert such a role. Interestingly, PLC is activated by hepatocellular swelling (1,32). The present study was therefore designed to explore the role of the PIP 2 /PLC signaling pathway in the modulation of resting and swelling-activated KCNQ1-like K ϩ currents in short-term cultured rat hepatocytes and to determine whether these channels contribute to RVD-induced whole organ K ϩ flux in the intact liver.…”

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confidence: 99%

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Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Wang¹,

Hill²

2006

American Journal of Physiology-Cell Physiology

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K+ channels participate in the regulatory volume decrease (RVD) accompanying hepatocellular nutrient uptake and bile formation. We recently identified KCNQ1 as a molecular candidate for a significant fraction of the hepatocellular swelling-activated K+ current ( IKVol). We have shown that the KCNQ1 inhibitor chromanol 293B significantly inhibited RVD-associated K+ flux in isolated perfused rat liver and used patch-clamp techniques to define the signaling pathway linking swelling to IKVol activation. Patch-electrode dialysis of hepatocytes with solutions that maintain or increase phosphatidylinositol 4,5-bisphosphate (PIP2) increased IKVol, whereas conditions that decrease cellular PIP2 decreased IKVol. GTP and AlF4− stimulated IKVol development, suggesting a role for G proteins and phospholipase C (PLC). Supporting this, the PLC blocker U-73122 decreased IKVol and inhibited the stimulatory response to PIP2 or GTP. Protein kinase C (PKC) is involved, because K+ current was enhanced by 1-oleoyl-2-acetyl- sn-glycerol and inhibited after chronic PKC stimulation with phorbol 12-myristate 13-acetate (PMA) or the PKC inhibitor GF 109203X. Both IKVol and the accompanying membrane capacitance increase were blocked by cytochalasin D or GF 109203X. Acute PMA did not eliminate the cytochalasin D inhibition, suggesting that PKC-mediated IKVol activation involves the cytoskeleton. Under isotonic conditions, a slowly developing K+ current similar to IKVol was activated by PIP2, lipid phosphatase inhibitors to counter PIP2 depletion, a PLC-coupled α1-adrenoceptor agonist, or PKC activators and was depressed by PKC inhibition, suggesting that hypotonicity is one of a set of stimuli that can activate IKVol through a PIP2/PKC-dependent pathway. The results indicate that PIP2 indirectly activates hepatocellular KCNQ1-like channels via cytoskeletal rearrangement involving PKC activation.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Wang¹,

Hill²

2006

American Journal of Physiology-Cell Physiology

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“…There are several evidences for the role of tyrosine kinases in RVD (8). The nonreceptor tyrosine kinases Src and focal adhesion kinase (FAK) has previously been reported to be activated during hypotonic stimulation in various cell types; i.e., phosphorylation of an activation site in FAK have previously been reported in, for example, chicken retina cells (9), human umbilical vein endothelial cells (15), and in cerebellar granule neurons (30), whereas hypotonic activation of Src has been reported in cerebellar granule neurons (30), rat hepatocytes (45), and in rat hepatoma cells (2). There is however no clear evidence linking the activation of tyrosine kinases to activation of swelling-activated K ϩ currents (I Kvol ).…”

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confidence: 99%

Activation of the TASK-2 channel after cell swelling is dependent on tyrosine phosphorylation

Kirkegaard

Lambert

Gammeltoft

et al. 2010

American Journal of Physiology-Cell Physiology

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Kirkegaard SS, Lambert IH, Gammeltoft S, Hoffmann EK. Activation of the TASK-2 channel after cell swelling is dependent on tyrosine phosphorylation. Am J Physiol Cell Physiol 299: C844 -C853, 2010. First published July 14, 2010; doi:10.1152/ajpcell.00024.2010.-The swelling-activated K ϩ currents (I K,vol ) in Ehrlich ascites tumor cells (EATC) has been reported to be through the two-pore domain (K2p), TWIK-related acid-sensitive K ϩ channel 2 (TASK-2). The regulatory volume decrease (RVD), following hypotonic exposure in EATC, is rate limited by IK,vol indicating that inhibition of RVD reflects inhibition of TASK-2. We find that in EATC the tyrosine kinase inhibitor genistein inhibits RVD by 90%, and that the tyrosine phosphatase inhibitor monoperoxo(picolinato)-oxo-vanadate(V) [mpV(pic)] shifted the volume set point for inactivation of the channel to a lower cell volume. Swelling-activated K ϩ efflux was impaired by genistein and the Src kinase family inhibitor 4-amino-5-(4-chloro-phenyl)-7-(t-butyl) pyrazolo [3,4-d]pyrimidine (PP2) and enhanced by the tyrosine phosphatase inhibitor mpV(pic). With the use of the TASK-2 inhibitor clofilium, it is demonstrated that mpV(pic) increased the volumesensitive part of the K ϩ efflux 1.3 times. To exclude K ϩ efflux via a KCl cotransporter, cellular Cl Ϫ was substituted with NO 3 Ϫ . Also under these conditions K ϩ efflux was completely blocked by genistein. Thus tyrosine kinases seem to be involved in the activation of the volumesensitive K ϩ channel, whereas tyrosine phosphatases appears to be involved in inactivation of the channel. Overexpressing TASK-2 in human embryonic kidney (HEK)-293 cells increased the RVD rate and reduced the volume set point. TASK-2 has tyrosine sites, and precipitation of TASK-2 together with Western blotting and antibodies against phosphotyrosines revealed a cell swelling-induced, timedependent tyrosine phosphorylation of the channel. Even though we found an inhibiting effect of PP2 on RVD, neither Src nor the focal adhesion kinase (FAK) seem to be involved. Inhibitors of the epidermal growth factor receptor tyrosine kinases had no effect on RVD, whereas the Janus kinase (JAK) inhibitor cucurbitacin inhibited the RVD by 40%. It is suggested that the cytokine receptor-coupled JAK/STAT pathway is upstream of the swelling-induced phosphorylation and activation of TASK-2 in EATC.cell volume regulation; Janus kinase; volume-sensitive channels

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“…Previous studies demonstrated that Src kinase was required for integrin-mediated VRAC activation (Browe and Baumgarten, 2003;Barfod et al, 2005 increased Cl − current and Cl − efflux induced by hypotonic solution . We hypothesized that integrin β3 mediates ICl.vol through Src and ClC-3.…”

Section: Integrin β3 Mediated Vrac Through the Src/clc-3 Signalling Pmentioning

confidence: 95%

Integrin β3 mediates cerebrovascular remodelling through Src/ClC‐3 volume‐regulated Cl⁻ channel signalling pathway

Zeng

Wang

et al. 2014

British J Pharmacology

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Background and PurposeCerebrovascular remodelling is one of the important risk factors of stroke. The underlying mechanisms are unclear. Integrin β3 and volume‐regulated ClC‐3 Cl− channels have recently been implicated as important contributors to vascular cell proliferation. Therefore, we investigated the role of integrin β3 in cerebrovascular remodelling and related Cl− signalling pathway.Experimental ApproachCl− currents were recorded using a patch clamp technique. The expression of integrin β3 in hypertensive animals was examined by Western blot and immunohistochemisty. Immunoprecipitation, cDNA and siRNA transfection were employed to investigate the integrin β3/Src/ClC‐3 signalling.Key ResultsIntegrin β3 expression was up‐regulated in stroke‐prone spontaneously hypertensive rats, 2‐kidney 2‐clip hypertensive rats and angiotensin II‐infused hypertensive mice. Integrin β3 expression was positively correlated with medial cross‐sectional area and ClC‐3 expression in the basilar artery of 2‐kidney 2‐clip hypertensive rats. Knockdown of integrin β3 inhibited the proliferation of rat basilar vascular smooth muscle cells induced by angiotensin II. Co‐immunoprecipitation and immunofluorescence experiments revealed a physical interaction between integrin β3, Src and ClC‐3 protein. The integrin β3/Src/ClC‐3 signalling pathway was shown to be involved in the activation of volume‐regulated chloride channels induced by both hypo‐osmotic stress and angiotensin II. Tyrosine 284 within a concensus Src phosphorylation site was the key point for ClC‐3 channel activation. ClC‐3 knockout significantly attenuated angiotensin II‐induced cerebrovascular remodelling.Conclusions and ImplicationsIntegrin β3 mediates cerebrovascular remodelling during hypertension via Src/ClC‐3 signalling pathway.

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Src Regulates Distinct Pathways for Cell Volume Control through Vav andPhospholipaseCγ

Cited by 24 publications

References 59 publications

Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Activation of the TASK-2 channel after cell swelling is dependent on tyrosine phosphorylation

Integrin β3 mediates cerebrovascular remodelling through Src/ClC‐3 volume‐regulated Cl⁻ channel signalling pathway

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Src Regulates Distinct Pathways for Cell Volume Control through Vav andPhospholipaseCγ

Cited by 24 publications

References 59 publications

Modulation of hepatocellular swelling-activated K+ currents by phosphoinositide pathway-dependent protein kinase C

Modulation of hepatocellular swelling-activated K+ currents by phosphoinositide pathway-dependent protein kinase C

Activation of the TASK-2 channel after cell swelling is dependent on tyrosine phosphorylation

Integrin β3 mediates cerebrovascular remodelling through Src/ClC‐3 volume‐regulated Cl− channel signalling pathway

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Product

Resources

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Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Modulation of hepatocellular swelling-activated K⁺ currents by phosphoinositide pathway-dependent protein kinase C

Integrin β3 mediates cerebrovascular remodelling through Src/ClC‐3 volume‐regulated Cl⁻ channel signalling pathway