scholarly journals Adenosine stimulates Ca2+ fluxes and increases cytosolic free Ca2+ in cultured rat mesangial cells

1992 ◽  
Vol 282 (3) ◽  
pp. 871-876 ◽  
Author(s):  
A Olivera ◽  
A López-Rivas ◽  
J M López-Novoa
Keyword(s):  
Mesangial Cells ◽  
Cell Types ◽  
Receptor Subtype ◽  
Glomerular Mesangial Cells ◽  
A1 Adenosine Receptor ◽  
Equilibrium Conditions ◽  
Intracellular Ca ◽  
Lower Magnitude ◽  
Initial Peak ◽  
Ca2 Channels

Adenosine has been associated with cellular Ca2+ metabolism in some cell types. Since adenosine is able to contract glomerular mesangial cells in culture, and since Ca2+ is the main messenger mediating contractile responses, we studied the effect of adenosine on 45Ca2+ movements into and out of mesangial cells and on the cytosolic free Ca2+ concentration ([Ca2+]i). Adenosine at 0.1 mM increased 45Ca2+ uptake (basal, 9993 +/- 216; + adenosine, 14823 +/- 410 d.p.m./mg; P less than 0.01) through verapamil-sensitive Ca2+ channels. These channels seem to be of the A1-adenosine receptor subtype. Adenosine also stimulated 45Ca2+ efflux from 45Ca(2+)-loaded mesangial cells. This effect was accompanied by a net depletion of intracellular 45Ca2+ content under isotopic equilibrium conditions (basal, 24213 +/- 978; + adenosine, 18622 +/- 885 d.p.m./mg; P less than 0.05). The increase in 45Ca2+ efflux was inhibited by a Ca(2+)-free medium or in the presence of 10 microM-verapamil. However, the intracellular Ca(2+)-release blocker TMB-8 (10 microM) only partially inhibited the adenosine-stimulated 45Ca2+ efflux. In addition, adenosine induced an elevation in [Ca2+]i in mesangial cells with an initial transient peak within 15 s (basal, 113 +/- 7; adenosine, 345 +/- 46 nM), and a secondary increase which was slower (3-4 min) and of lower magnitude than the initial peak (250 +/- 21 nM). In summary, adenosine elevates [Ca2+]i and stimulates both Ca2+ uptake from the extracellular pool and Ca2+ efflux from intracellular pools in mesangial cells. The Ca2+ release from internal stores is produced by a combination of a TMB-8-inhibitable and a non-TMB-8-inhibitable mechanism, and seems to be dependent on Ca2+ influx.

Ca2+ channels mediate protein tyrosine kinase activation by endothelin-1

1996 ◽  
Vol 270 (5) ◽  
pp. F790-F797 ◽  
Author(s):  
M. S. Simonson ◽  
Y. Wang ◽  
W. H. Herman
Keyword(s):  
Tyrosine Phosphorylation ◽  
Tyrosine Kinases ◽  
Mesangial Cells ◽  
Focal Adhesions ◽  
Cellular Protein ◽  
Early Gene ◽  
Glomerular Mesangial Cells ◽  
Protein Tyrosine ◽  
Endothelin 1 ◽  
Ca2 Channels

To investigate the novel interaction between endothelin-1 (ET-1) and cellular protein tyrosine kinases (PTK), we asked whether Ca2+ influx links ET-1 receptors to PTK activation. In glomerular mesangial cells, ET-1 stimulated a biphasic increase in PTK activity in anti-phosphotyrosine immunoprecipitates that temporally correlated with increased tyrosine phosphorylation of cellular proteins. ET-1 increased tyrosine phosphorylation of proteins in the cytosol and in a puncture distribution consistent with focal adhesions. Addition of ionomycin to increase Ca2+ influx stimulated PTK activity, and inhibition of extracellular Ca2+ influx blocked PTK activation by ET-1. ET-1 increased autophosphorylation of pp60c-src, which was mimicked by addition of ionomycin and inhibited by chelation of extracellular Ca2+. In addition, a selective PTK inhibitor blocked induction of c-fos mRNA by ionomycin, suggesting that Ca(2+)-stimulated PTKs contribute to a signaling pathway regulating immediate early gene expression. Taken together, these results demonstrate that ET-1 stimulates nonreceptor PTK activity, including pp60c-src, by activating Ca2+ channels and subsequent influx of extracellular Ca2+.

TRPC4 forms store-operated Ca2+ channels in mouse mesangial cells

2004 ◽  
Vol 287 (2) ◽  
pp. C357-C364 ◽  
Author(s):  
Xiaoxia Wang ◽  
Jennifer L. Pluznick ◽  
Peilin Wei ◽  
Babu J. Padanilam ◽  
Steven C. Sansom
Keyword(s):  
Transient Receptor Potential ◽  
Mesangial Cells ◽  
Receptor Potential ◽  
Immunocytochemical Staining ◽  
Rt Pcr ◽  
Fura 2 ◽  
Intracellular Ca ◽  
Transient Receptor ◽  
Ca2 Channels

Studies were performed to identify the molecular component responsible for store-operated Ca2+ entry in murine mesangial cells (MMC). Because the canonical transient receptor potential (TRPC) family of proteins was previously shown to comprise Ca2+-selective and -nonselective cation channels in a variety of cells, we screened TRPC1–TRPC7 with the use of molecular methods and the fura 2 method to determine their participation as components of the mesangial store-operated Ca2+ (SOC) channel. Using TRPC-specific primers and RT-PCR, we found that cultured MMC contained mRNA for TRPC1 and TRPC4 but not for TRPC2, TRPC3, TRPC5, TRPC6, and TRPC7. Immunocytochemical staining of MMC revealed predominantly cytoplasmic expression of TRPC1 and plasmalemmal expression of TRPC4. The role of TRPC4 in SOC was determined with TRPC4 antisense and fura 2 ratiometric measurements of intracellular Ca2+ concentration ([Ca2+]i). SOC was measured as the increase in [Ca2+]i after extracellular Ca2+ was increased from <10 nM to 1 mM in the continued presence of thapsigargin. We found that TRPC4 antisense, which reduced plasmalemmal expression of TRPC4, inhibited SOC by 83%. Incubation with scrambled TRPC4 oligonucleotides did not affect SOC. Immunohistochemical staining identified expressed TRPC4 in the glomeruli of mouse renal sections. The results of RT-PCR performed to distinguish between TRPC4-α and TRPC4-β were consistent with expression of both isoforms in brain but with only TRPC4-α expression in MMC. These studies show that TRPC4-α may form the homotetrameric SOC in mouse mesangial cells.

Regulation of mesangial cell cyclooxygenase synthesis by cytokines and glucocorticoids

1992 ◽  
Vol 263 (1) ◽  
pp. F97-F102 ◽  
Author(s):  
D. W. Coyne ◽  
M. Nickols ◽  
W. Bertrand ◽  
A. R. Morrison
Keyword(s):  
Time Course ◽  
Mesangial Cell ◽  
Mesangial Cells ◽  
Interleukin 1 ◽  
Cell Types ◽  
Dependent Manner ◽  
Glomerular Mesangial Cells ◽  
Prostaglandin Production ◽  
Arachidonate Release ◽  
Necrosis Factor

The cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF), potently induce prostaglandin formation in glomerular mesangial cells. Mechanisms by which these cytokines stimulate prostaglandin formation vary among cell types. We investigated whether alterations in phospholipase A2 (PLA2) or cyclooxygenase (COX) mass and activity contribute to the changes in mesangial cell prostaglandin production. These cytokines induced COX activity and mass in a time-dependent manner, which paralleled prostaglandin production. IL-1 increased COX mass approximately threefold by 24 h. TNF had a much smaller effect, although it appeared to be additive with IL-1. IL-1-induced COX mass was maintained at an increased level for at least 48 h. The glucocorticoid dexamethasone (DEX) virtually abolished prostaglandin production and blocked cytokine induction of COX activity and mass. DEX did not reduce COX activity or mass below the basal, serum-fed levels, however. By utilizing stable isotope methods, we could demonstrate that IL-1 increased free arachidonate levels, implying new PLA2 synthesis over a time course that was maximal at 6 h and was cycloheximide and actinomycin D sensitive. These data demonstrate that the cytokines IL-1 and TNF enhance synthesis of COX and PLA2, contributing to increased prostaglandin production. Cytokine-stimulated prostaglandin production ceases when cells are also treated with DEX, although control levels of COX activity and mass remain. This occurs because DEX inhibits the IL-1-induced enhanced arachidonate release.

scholarly journals Hyaluronan Is Not a Ligand but a Regulator of Toll-Like Receptor Signaling in Mesangial Cells: Role of Extracellular Matrix in Innate Immunity

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Rainer Ebid ◽  
Julia Lichtnekert ◽  
Hans-Joachim Anders
Keyword(s):  
Extracellular Matrix ◽  
Innate Immunity ◽  
Cell Surface ◽  
Mesangial Cells ◽  
Cell Types ◽  
Dependent Manner ◽  
Glomerular Mesangial Cells ◽  
Toll Like Receptor ◽  
Surface Receptors ◽  
Tlr4 Ligand

Glomerular mesangial cells (MC), like most cell types secrete hyaluronan (HA), which attached to the cell surface via CD44, is the backbone of a hydrophilic gel matrix around these cells. Reduced extracellular matrix thickness and viscosity result from HA cleavage during inflammation. HA fragments were reported to trigger innate immunity via Toll-like receptor-(TLR-) 2 and/or TLR4 in immune cells. We questioned whether HA fragments also regulate the immunostimulatory capacity of smooth muscle cell-like MC. LPS (TLR4-ligand) and PAM3CysSK4 (TLR2-ligand) induced IL-6 secretion in MC; highly purified endotoxin-free HA < 3000 Da up to 50 μg/mL did not. Bovine-testis-hyaluronidase from was used to digest MC-HA into HA fragments of different size directly in the cell culture. Resultant HA fragments did not activate TLR4-deficient MC, while TLR2-deficient MC responded to LPS-contamination of hyaluronidase, not to produced HA fragments. Hyaluronidase increased the stimulatory effect of TLR2-/-3/-5 ligands on their TLR-receptors in TLR4-deficient MC, excluding any effect by LPS-contamination. Supplemented heparin suppressed every stimulatory effect in a dose-dependent manner. We conclude that the glycosaminoglycan HA creates a pericellular jelly barrier, which covers surface receptors like the TLRs. Barrier-thickness and viscosity balanced by HA-synthesis and degradation and the amount of HA-receptors on the cell surface regulate innate immunity via the accessibility of the receptors.

scholarly journals RCAN1.4 mediates high glucose-induced matrix production by stimulating mitochondrial fission in mesangial cells

2020 ◽  
Vol 40 (1) ◽  
Author(s):  
Hong-Min Chen ◽  
Jia-Jia Dai ◽  
Rui Zhu ◽  
Xue-Yu Sang ◽  
Fang-Fang Peng ◽  
...  
Keyword(s):  
High Glucose ◽  
Matrix Protein ◽  
Mesangial Cells ◽  
Mitochondrial Dynamics ◽  
Mitochondrial Fission ◽  
Cell Types ◽  
Glomerular Mesangial Cells ◽  
Mitochondrial Fragmentation ◽  
Mitochondrial Dynamic ◽  
Matrix Production

Abstract High glucose (HG)-induced mitochondrial dynamic changes and oxidative damage are closely related to the development and progression of diabetic kidney disease (DKD). Recent studies suggest that regulators of calcineurin 1 (RCAN1) is involved in the regulation of mitochondrial function in different cell types, so we investigate the role of RCAN1 in mitochondrial dynamics under HG ambience in rat glomerular mesangial cells (MCs). MCs subjected to HG exhibited an isoform-specific up-regulation of RCAN1.4 at both mRNA and protein levels. RCAN1.4 overexpression induced translocation of Dynamin related protein 1 (Drp1) to mitochondria, mitochondrial fragmentation and depolarization, accompanied by increased matrix production under normal glucose and HG ambience. In contrast, decreasing the expression of RCAN1.4 by siRNA inhibited HG-induced mitochondrial fragmentation and matrix protein up-regulation. Moreover, both mitochondrial fission inhibitor Mdivi-1 and Drp1 shRNA prevented RCAN1.4-induced fibronectin up-regulation, suggesting that RCAN1.4-induced matrix production is dependent on its modulation of mitochondrial fission. Although HG-induced RCAN1.4 up-regulation was achieved by activating calcineurin, RCAN1.4-mediated mitochondrial fragmentation and matrix production is independent of calcineurin activity. These results provide the first evidence for the HG-induced RCAN1.4 up-regulation involving increased mitochondrial fragmentation, leading to matrix protein up-regulation.

Unexpected protection of glomerular mesangial cells from oxidant-triggered apoptosis by bioflavonoid quercetin

1997 ◽  
Vol 273 (2) ◽  
pp. F206-F212 ◽  
Author(s):  
T. Yokoo ◽  
M. Kitamura
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Molecular Mechanisms ◽  
Kinase Inhibitors ◽  
Mesangial Cells ◽  
Oxidant Stress ◽  
Cell Types ◽  
Immunoblot Analysis ◽  
Dependent Manner ◽  
Glomerular Mesangial Cells

Bioflavonoid quercetin is known as an anti-cancer agent that induces apoptosis of tumor cells. Currently, however, little is understood about the effect of this drug on the function of normal cells. In this report, we address an unexpected, novel action of quercetin against apoptosis. Pretreatment with quercetin protected mesangial cells from hydrogen peroxide (H2O2)-induced apoptosis. A similar effect was observed in other cell types including LLC-PK1 epithelial cells and NRK49F fibroblasts. To explore the molecular mechanisms involved, we tested the effect of quercetin on c-Jun/activator protein-1 AP-1), the crucial mediator for H2O2-initiated apoptosis. Northern blot analysis revealed that quercetin suppressed the c-jun expression by H2O2. This was correlated with blunted activation of 12-O-tetradecanoylphorbol 13-acetate response element (TRE) in response to H2O2. These results suggested that quercetin inhibited apoptosis via intervention in the c-Jun/AP-1 pathway. To further investigate the action of quercetin, its effect on tyrosine kinases was studied. Immunoblot analysis revealed that H2O2 induced tyrosine phosphorylation. Quercetin inhibited this process in a dose-dependent manner. Inactivation of tyrosine kinases was an event upstream of c-Jun/AP-1, because tyrosine kinase inhibitors suppressed both activation of c-Jun/AP-1 and induction of apoptosis by H2O2. These findings elucidated the novel action of quercetin as an apoptosis inhibitor. This cytoprotective effect was found to be via suppression of the tyrosine kinase-c-Jun/AP-1 pathway triggered by oxidant stress.

Mediation of angiotensin II-induced Ca2+ signaling by polycystin 2 in glomerular mesangial cells

2008 ◽  
Vol 294 (4) ◽  
pp. F909-F918 ◽  
Author(s):  
Juan Du ◽  
Min Ding ◽  
Sherry Sours-Brothers ◽  
Sarabeth Graham ◽  
Rong Ma
Keyword(s):  
Transient Receptor Potential ◽  
Mesangial Cells ◽  
Rat Kidney ◽  
Receptor Potential ◽  
Cell Types ◽  
Protein Product ◽  
Dominant Negative ◽  
Ang Ii ◽  
Glomerular Mesangial Cells ◽  
Autosomal Dominant Polycystic Kidney

Ca+ influx across the plasma membrane is a major component of mesangial cell (MC) response to vasoconstrictors. Polycystin 2 (PC2), the protein product of the gene mutated in type 2 autosomal dominant polycystic kidney disease, has been shown to function as a nonselective cation channel in a variety of cell types. The present study was performed to test the hypothesis that PC2 and its binding partners constitute a Ca2+-permeable channel and contribute to ANG II-induced Ca2+ signaling in MCs. Western blot and immunocytochemistry showed PC2 expression in cultured human MCs. The existence of PC2 in MCs was further confirmed by immunohistochemsitry in rat kidney sections. Coimmunoprecipitation displayed a selective interaction of PC2 with canonical transient receptor potential (TRPC) proteins TRPC1 and TRPC4. Cell-attached patch-clamp experiments revealed that ANG II-induced membrane currents were enhanced by overexpression of pkd2 but significantly inhibited by knock down of pkd2, 30 μM Gd3+ (a PC2 channel blocker), and dominant-negative pkd2 mutant (pkd2-D511V). Corresponding to the increase in channel currents, ANG II stimulation increased expression of PC2 on the cell surface of MCs and interaction with TRPC1 and TRPC4. Furthermore, ANG II-induced MC contraction was significantly reduced in pkd2-knocked down MCs. These data suggest that PC2 selectively assembles with TRPC1 and TRPC4 to form channel complexes mediating ANG II-induced Ca2+ responses in MCs.

AVP-induced Ca fluxes and contraction of rat glomerular mesangial cells

1988 ◽  
Vol 255 (1) ◽  
pp. F142-F150 ◽  
Author(s):  
K. Takeda ◽  
H. Meyer-Lehnert ◽  
J. K. Kim ◽  
R. W. Schrier
Keyword(s):  
Channel Blocker ◽  
Mesangial Cells ◽  
Cytosolic Calcium ◽  
Glomerular Mesangial Cells ◽  
Free Medium ◽  
Cell Contraction ◽  
High K ◽  
Intracellular Ca ◽  
Lanthanum Ion ◽  
The Relationship

Arginine vasopressin (AVP) is known to exert Ca mobilization and contraction in glomerular mesangial cells and vascular smooth muscle cells. However, the relationship between changes in intracellular Ca and transmembrane Ca fluxes is not clear at the present time. Therefore, this study was undertaken to determine the effect of AVP on cytosolic calcium ([Ca2+]i) and Ca fluxes as estimated by measurements of 45Ca2+ efflux. Changes of [Ca2+]i in response to AVP were directly measured in monolayers of adherent cultured mesangial cells. AVP induced rapid concentration-dependent increases in [Ca2+]i and Ca2+ efflux. AVP also induced contraction of mesangial cells. This effect was blocked only by the V1 (pressor)-antagonist, d(CH2)5Tyr(Me)AVP. Stimulation of Ca2+ efflux and changes in [Ca2+]i by AVP completely desensitized the mesangial cells to a subsequent identical challenge of AVP with no cross-tachyphylaxis to other hormones. Even in Ca2+-free medium, AVP increased [Ca2+]i and Ca2+ efflux, but to a lesser extent. Under this condition, contraction of mesangial cells induced by AVP was also observed. Readdition of extracellular Ca2+ following the AVP-induced increase in [Ca2+]i caused a second and slower [Ca2+]i increase. In Ca2+-containing conditions, lanthanum ion-reduced AVP evoked [Ca2+]i stimulation to the value observed in Ca2+-free medium. The Ca2+ channel blocker, verapamil, partially inhibited AVP-induced Ca2+ influx but totally blocked the increase in [Ca2+]i induced by high K. Verapamil did not inhibit AVP-stimulated Ca2+ efflux and cell contraction. Dantrolene, a blocker of Ca2+ release from endoplasmic reticulum, inhibited AVP-stimulated Ca2+ efflux and cell contraction.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Functional Ca2+ Channels between Channel Clusters are Necessary for the Propagation of IP3R-Mediated Ca2+ Waves

2019 ◽  
Vol 24 (2) ◽  
pp. 61
Author(s):  
Estefanía Piegari ◽  
Silvina Ponce Dawson
Keyword(s):  
Signal Propagation ◽  
Cell Types ◽  
Quantitative Comparison ◽  
Preliminary Conclusion ◽  
Qualitative Comparison ◽  
Excitable System ◽  
Inositol 1,4,5 Trisphosphate ◽  
Intracellular Ca ◽  
Spatio Temporal ◽  
Ca2 Channels

The specificity and universality of intracellular Ca 2 + signals rely on the variety of spatio-temporal patterns that the Ca 2 + concentration can display. Ca 2 + release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP 3 Rs) is key for this variety. The opening probability of IP 3 Rs depends on the cytosolic Ca 2 + concentration. All of the dynamics are then well described by an excitable system in which the signal propagation depends on the ability of the Ca 2 + released through one IP 3 R to induce the opening of other IP 3 Rs. In most cell types, IP 3 Rs are organized in clusters, i.e., the cytosol is a “patchy” excitable system in which the signals can remain localized (i.e., involving the release through one or more IP 3 Rs in a cluster), or become global depending on the efficiency of the Ca 2 + -mediated coupling between clusters. The spatial range over which the signals propagate determines the responses that the cell eventually produces. This points to the importance of understanding the mechanisms that make the propagation possible. Our previous qualitative comparison between experiments and numerical simulations seemed to indicate that Ca 2 + release not only occurs within the close vicinity of the clearly identifiable release sites (IP 3 R clusters) but that there are also functional IP 3 Rs in between them. In this paper, we present a quantitative comparison between experiments and models that corroborate this preliminary conclusion. This result has implications on how the Ca 2 + -mediated coupling between clusters works and how it can eventually be disrupted by the different Ca 2 + trapping mechanisms.

Mesangial cell-reduced Ca2+ signaling in high glucose is due to inactivation of phospholipase C-β3 by protein kinase C

2005 ◽  
Vol 289 (5) ◽  
pp. F1078-F1087 ◽  
Author(s):  
Helena Frecker ◽  
Snezana Munk ◽  
Hong Wang ◽  
Catharine Whiteside
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phospholipase C ◽  
High Glucose ◽  
Mesangial Cells ◽  
Fold Increase ◽  
Glomerular Mesangial Cells ◽  
Kinase C ◽  
Normal Glucose ◽  
Intracellular Ca

In high glucose, glomerular mesangial cells (MCs) demonstrate impaired Ca2+ signaling in response to seven-transmembrane receptor stimulation. To identify the mechanism, we first postulated decreased release from intracellular stores. Intracellular Ca2+ was measured in fluo-3-loaded primary cultured rat MCs using confocal fluorescence microscopy. In high glucose (HG) 30 mM for 48 h, the 25 nM ionomycin-stimulated intracellular Ca2+ response was reduced to 82% of that observed in normal glucose (NG). In NG 5.6 mM, Ca2+ responses to endothelin (ET)-1 and platelet-derived growth factor (PDGF) were unchanged in cells cultured in 50 nM Ca2+ vs. 1.8 mM Ca2+. Depletion of intracellular Ca2+ stores with thapsigargin eliminated ET-1-stimulated Ca2+ responses. Incubation in 30 mM glucose (HG) for 48 h or stimulation with phorbol myristate acetate (PMA) for 10 min eliminated the Ca2+ response to ET-1 but had no effect on the PDGF response. Downregulation of protein kinase C (PKC) with 24-h PMA or inhibition with Gö6976 in HG normalized the Ca2+ response to ET-1. Because ET-1 and PDGF stimulate Ca2+ signaling through different phospholipase C pathways, we hypothesized that, in HG, PKC selectively phosphorylates and inhibits PLC-β3. Using confocal immunofluorescence imaging, in NG, a 1.6- to 1.7-fold increase in PLC-β3 Ser1105 phosphorylation was observed following PMA or ET-1 stimulation for 10 min. In HG, immunofluorescent imaging and immunoblotting showed increased PLC-β3 phosphorylation, without change in total PLC-β3, which was reversed with 24-h PMA or Gö6976. We conclude that reduced Ca2+ signaling in HG cannot be explained by reduced Ca2+ stores but is due to conventional PKC-dependent phosphorylation and inactivation of PLC-β3.

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