Effects of osmotic stress on the activity of MAPKs and PDGFR-β-mediated signal transduction in NIH-3T3 fibroblasts

2008 ◽  
Vol 294 (4) ◽  
pp. C1046-C1055 ◽  
Author(s):  
M.-B. Nielsen ◽  
S. T. Christensen ◽  
E. K. Hoffmann
Keyword(s):  
Signal Transduction ◽  
Osmotic Stress ◽  
Cell Volume ◽  
Nuclear Translocation ◽  
Cell Swelling ◽  
Cell Shrinkage ◽  
Jnk Pathway ◽  
Mapk Activity ◽  
3T3 Fibroblasts ◽  
Nih 3T3

Signaling in cell proliferation, cell migration, and apoptosis is highly affected by osmotic stress and changes in cell volume, although the mechanisms underlying the significance of cell volume as a signal in cell growth and death are poorly understood. In this study, we used NIH-3T3 fibroblasts in a serum- and nutrient-free inorganic medium (300 mosM) to analyze the effects of osmotic stress on MAPK activity and PDGF receptor (PDGFR)-β-mediated signal transduction. We found that hypoosmolarity (cell swelling at 211 mosM) induced the phosphorylation and nuclear translocation of ERK1/2, most likely via a pathway independent of PDGFR-β and MEK1/2. Conversely, hyperosmolarity (cell shrinkage at 582 mosM) moved nuclear and phosphorylated ERK1/2 to the cytoplasm and induced the phosphorylation and nuclear translocation of p38 and phosphorylation of JNK1/2. In a series of parallel experiments, hypoosmolarity did not affect PDGF-BB-induced activation of PDGFR-β, whereas hyperosmolarity strongly inhibited ligand-dependent PDGFR-β activation as well as downstream mitogenic signal components of the receptor, including Akt and the MEK1/2-ERK1/2 pathway. Based on these results, we conclude that ligand-dependent activation of PDGFR-β and its downstream effectors Akt, MEK1/2, and ERK1/2 is strongly modulated (inhibited) by hyperosmotic cell shrinkage, whereas cell swelling does not seem to affect the activation of the receptor but rather to activate ERK1/2 via a different mechanism. It is thus likely that cell swelling via activation of ERK1/2 and cell shrinkage via activation of the p38 and JNK pathway and inhibition of the PDGFR signaling pathway may act as key players in the regulation of tissue homeostasis.

Regulation of cell function by the cellular hydration state

1994 ◽  
Vol 267 (3) ◽  
pp. E343-E355 ◽  
Author(s):  
D. Haussinger ◽  
F. Lang ◽  
W. Gerok
Keyword(s):  
Cell Volume ◽  
Cell Function ◽  
Narrow Range ◽  
Cell Swelling ◽  
Metabolic Function ◽  
Cell Shrinkage ◽  
Short Term ◽  
Hydration State ◽  
Per Se ◽  
And Oxidative Stress

Cellular hydration can change within minutes under the influence of hormones, nutrients, and oxidative stress. Such short-term modulation of cell volume within a narrow range acts per se as a potent signal which modifies cellular metabolism and gene expression. It appears that cell swelling and cell shrinkage lead to certain opposite patterns of cellular metabolic function. Apparently, hormones and amino acids can trigger those patterns simply by altering cell volume. Thus alterations of cellular hydration may represent another important mechanism for metabolic control and act as another second or third messenger linking cell function to hormonal and environmental alterations.

Volume-sensitive Ca influx and release from intracellular pools in gastric parietal cells

1992 ◽  
Vol 263 (3) ◽  
pp. C584-C589 ◽  
Author(s):  
P. A. Negulescu ◽  
B. Munck ◽  
T. E. Machen
Keyword(s):  
Image Processing ◽  
Cell Volume ◽  
Digital Image Processing ◽  
Parietal Cells ◽  
Cell Swelling ◽  
Cell Shrinkage ◽  
Gastric Glands ◽  
Intact Rabbit ◽  
Induced Changes ◽  
Gastric Parietal Cells

The effects of osmotically induced changes in cell volume on cytoplasmic free Ca (Cai) were studied in parietal cells from intact rabbit gastric glands using digital image processing of fura-2 fluorescence. In resting unstimulated cells, Cai was unaffected by either cell swelling or shrinking when osmolarity was varied between 200 and 400 mosM (isotonicity 290 mosM). However, when cells were swelled in a 165 mosM solution (55% tonicity), a biphasic Ca increased was observed. On average, Cai increased transiently from 80 to 218 nM before stabilizing at approximately 140 nM. The peak was due to release from intracellular pools because it was present in Ca-free solutions while the sustained elevation was dependent on external Ca. In carbachol-stimulated cells, Ca influx was most sensitive to cell shrinkage. For example, addition of 25 mM sucrose (108% tonicity) caused a 30% decrease in the sustained carbachol-stimulated Cai increase (plateau). In contrast, carbachol-stimulated cells were relatively insensitive to cell swelling, with a 30% decrease in tonicity causing only a 15% increase in the plateau. However, as in the unstimulated cells, extreme (55% tonicity) swelling caused additional increases in Cai levels. The carbachol-dependent effects of changes in cell volume on Cai could be mimicked by treating cells with thapsigargin, an inhibitor of Ca pumps of intracellular membranes that also has been shown to stimulate Ca entry. Thus, although extreme swelling conditions (55% tonicity) could elicit Cai increases in either the presence or absence of agonist, agonist was required to observe Cai decreases due to cell shrinkage.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effects of bradykinin on cell volume and intracellular pH in NIH 3T3 fibroblasts expressing the ras oncogene

FEBS Letters ◽  
1992 ◽  
Vol 307 (3) ◽  
pp. 367-370 ◽  
Author(s):  
M. Ritter ◽  
E. Wöll ◽  
D. Häussinger ◽  
F. Lang
Keyword(s):  
Cell Volume ◽  
Intracellular Ph ◽  
Ras Oncogene ◽  
3T3 Fibroblasts ◽  
Nih 3T3

scholarly journals Cell volume and bile acid excretion

1992 ◽  
Vol 288 (2) ◽  
pp. 681-689 ◽  
Author(s):  
D Häussinger ◽  
C Hallbrucker ◽  
N Saha ◽  
F Lang ◽  
W Gerok
Keyword(s):  
Amino Acids ◽  
Cell Volume ◽  
Liver Cell ◽  
Bile Flow ◽  
Cell Swelling ◽  
Isolated Perfused Rat Liver ◽  
Cell Shrinkage ◽  
Volume Changes ◽  
Close Relationship ◽  
Stimulation Of

The interaction between cell volume and taurocholate excretion into bile was studied in isolated perfused rat liver. Cell swelling due to hypo-osmotic exposure, addition of amino acids or insulin stimulated taurocholate excretion into bile and bile flow, whereas hyperosmotic cell shrinkage inhibited these. These effects were explained by changes in Vmax of taurocholate excretion into bile: Vmax. increased from about 300 to 700 nmol/min per g after cell swelling by 12-15% caused by either hypo-osmotic exposure or addition of amino acids under normo-osmotic conditions. Steady-state taurocholate excretion into bile was not affected when the influent K+ concentration was increased from 6 to 46 mM or decreased to 1 mM with iso-osmoticity being maintained by corresponding changes in the influent Na+ concentration. Replacement of 40 mM-NaCl by 80 mM-sucrose decreased taurocholate excretion into bile by about 70%; subsequent hypo-osmotic exposure by omission of sucrose increased taurocholate excretion to 160%. Only minor, statistically insignificant, effects of aniso-osmotic cell volume changes on the appearance of bolus-injected horseradish peroxidase in bile were observed. Taurocholate (400 microM) exhibited a cholestatic effect during hyperosmotic cell shrinkage, but not during hypo-osmotic cell swelling. Both taurocholate and tauroursodeoxycholate increased liver cell volume. Tauroursodeoxycholate stimulated taurocholate (100 microM) excretion into bile. This stimulatory effect was strongly dependent on the extent of tauroursodeoxycholate-induced cell swelling. During continuous infusion of taurocholate (100 microM) further addition of tauroursodeoxycholate at concentrations of 20, 50 and 100 microM increased cell volume by 10, 8 and 2% respectively, in parallel with a stimulation of taurocholate excretion into bile by 29, 27 and 9% respectively. There was a close relationship between the extent of cell volume changes and taurocholate excretion into bile, regardless of whether cell volume was modified by tauroursodeoxycholate, amino acids or aniso-osmotic exposure. The data suggest that: (i) liver cell volume is one important factor determining bile flow and biliary taurocholate excretion; (ii) swelling-induced stimulation of taurocholate excretion into bile is probably not explained by alterations of the membrane potential; (iii) bile acids modulate liver cell volume; (iv) taurocholate-induced cholestasis may depend on cell volume; (v) stimulation of taurocholate excretion into bile by tauroursodeoxycholate can largely be explained by tauroursodeoxycholate-induced cell swelling.

scholarly journals Stat6 and Jak1 Are Common Elements in Platelet-derived Growth Factor and Interleukin-4 Signal Transduction Pathways in NIH 3T3 Fibroblasts

1996 ◽  
Vol 271 (36) ◽  
pp. 22175-22182 ◽  
Author(s):  
Bharvin K. R. Patel ◽  
Ling-Mei Wang ◽  
Chong-Chou Lee ◽  
William G. Taylor ◽  
Jacalyn H. Pierce ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Growth Factor ◽  
Platelet Derived Growth Factor ◽  
Interleukin 4 ◽  
Signal Transduction Pathways ◽  
Common Elements ◽  
3T3 Fibroblasts ◽  
Nih 3T3

scholarly journals Cells recognize osmotic stress through liquid-liquid phase separation lubricated with poly(ADP-ribose)

2020 ◽  
Author(s):  
Kengo Watanabe ◽  
Kazuhiro Morishita ◽  
Xiangyu Zhou ◽  
Shigeru Shiizaki ◽  
Yasuo Uchiyama ◽  
...  
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Osmotic Stress ◽  
Cell Volume ◽  
Hyperosmotic Stress ◽  
Cell Swelling ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Genome Wide ◽  
A Genome ◽  
Interfering Rna

AbstractCells are under threat of osmotic perturbation; and cell volume maintenance is critical in cerebral edema, inflammation and aging, in which prominent changes in intracellular or extracellular osmolality emerge. After osmotic stress-enforced cell swelling or shrinkage, the cells regulate intracellular osmolality to recover their volume. However, the mechanisms recognizing osmotic stress remain obscured. We previously clarified that apoptosis signal-regulating kinase 3 (ASK3) bidirectionally responds to osmotic stress and regulates cell volume recovery. Here, we report that macromolecular crowding induces liquid-demixing condensates of ASK3 under hyperosmotic stress, which transduce osmosensing signal into ASK3 inactivation. A genome-wide small interfering RNA (siRNA) screen identified an ASK3 inactivation regulator, nicotinamide phosphoribosyltransferase (NAMPT), related to poly(ADP-ribose) signaling. Furthermore, we clarify that poly(ADP-ribose) keeps ASK3 condensates in the liquid phase and enables ASK3 to become inactivated under hyperosmotic stress. Our findings demonstrate that cells rationally incorporate physicochemical phase separation into their osmosensing systems.

scholarly journals Resting and osmotically induced basolateral K conductances in turtle colon.

1986 ◽  
Vol 88 (2) ◽  
pp. 253-274 ◽  
Author(s):  
W J Germann ◽  
S A Ernst ◽  
D C Dawson
Keyword(s):  
Epithelial Cells ◽  
Osmotic Stress ◽  
Cell Volume ◽  
Organic Anion ◽  
Basolateral Membrane ◽  
Cell Swelling ◽  
Na Transport ◽  
Colonic Epithelial Cells ◽  
Cell Layers ◽  
Normal Transport

Two types of K conductance can be distinguished in the basolateral membranes of polyene-treated colonic epithelial cells (see Germann, W. J., M. E. Lowy, S. A. Ernst, and D. C. Dawson, 1986, Journal of General Physiology, 88:237-251). The significance of these two types of K conductance was investigated by measuring the properties of the basolateral membrane under conditions that we presumed would lead to marked swelling of the epithelial cells. We compared the basolateral conductance under these conditions of osmotic stress with those observed under other conditions where changes in cell volume would be expected to be less dramatic. In the presence of a permeant salt (KCl) or nonelectrolyte (urea), amphotericin-treated colonic cell layers exhibited a quinidine-sensitive conductance. Light microscopy revealed that these conditions were also associated with pronounced swelling of the epithelial cells. Incubation of tissues in solutions containing the organic anion benzene sulfonate led to the activation of the quinidine-sensitive gK and was also associated with dramatic cell swelling. In contrast, tissues incubated with an impermeant salt (K-gluconate) or nonelectrolyte (sucrose) did not exhibit a quinidine-sensitive basolateral conductance in the presence of the polyene. Although such conditions were also associated with changes in cell volume, they did not lead to the extreme cell swelling detected under conditions that activated the quinidine-sensitive gK. The quinidine-sensitive basolateral conductance that was activated under conditions of osmotic stress was also highly selective for K over Rb, in contrast to the behavior of normal Na transport by the tissue, which was supported equally well by K or Rb and was relatively insensitive to quinidine. The results are consistent with the notion that the basolateral K conductance measured in the amphotericin-treated epithelium bathed by mucosal K-gluconate solutions or in the presence of sucrose was due to the same channels that are responsible for the basolateral K conductance under conditions of normal transport. Conditions of extreme osmotic stress, however, which led to pronounced swelling of the epithelial cells, were associated with the activation of a new conductance, which was highly selective for K over Rb and was blocked by quinidine or lidocaine.

scholarly journals WNK3 and WNK4 exhibit opposite sensitivity with respect to cell volume and intracellular chloride concentration

2020 ◽  
Vol 319 (2) ◽  
pp. C371-C380
Author(s):  
Diana Pacheco-Alvarez ◽  
Diego Luis Carrillo-Pérez ◽  
Adriana Mercado ◽  
Karla Leyva-Ríos ◽  
Erika Moreno ◽  
...  
Keyword(s):  
Cell Volume ◽  
Chloride Concentration ◽  
Cell Swelling ◽  
Cell Shrinkage ◽  
Serine Threonine Kinase ◽  
Intracellular Chloride ◽  
Highly Sensitive ◽  
A Cell ◽  
Carboxy Terminal ◽  
Intracellular Chloride Concentration

Cation-coupled chloride cotransporters (CCC) play a role in modulating intracellular chloride concentration ([Cl−]i) and cell volume. Cell shrinkage and cell swelling are accompanied by an increase or decrease in [Cl−]i, respectively. Cell shrinkage and a decrease in [Cl−]i increase the activity of NKCCs (Na-K-Cl cotransporters: NKCC1, NKCC2, and Na-Cl) and inhibit the activity of KCCs (K-Cl cotransporters: KCC1 to KCC4), wheras cell swelling and an increase in [Cl−]i activate KCCs and inhibit NKCCs; thus, it is unlikely that the same kinase is responsible for both effects. WNK1 and WNK4 are chloride-sensitive kinases that modulate the activity of CCC in response to changes in [Cl−]i. Here, we showed that WNK3, another member of the serine-threonine kinase WNK family with known effects on CCC, is not sensitive to [Cl−]i but can be regulated by changes in extracellular tonicity. In contrast, WNK4 is highly sensitive to [Cl−]i but is not regulated by changes in cell volume. The activity of WNK3 toward NaCl cotransporter is not affected by eliminating the chloride-binding site of WNK3, further confirming that the kinase is not sensitive to chloride. Chimeric WNK3/WNK4 proteins were produced, and analysis of the chimeras suggests that sequences within the WNK’s carboxy-terminal end may modulate the chloride affinity. We propose that WNK3 is a cell volume-sensitive kinase that translates changes in cell volume into phosphorylation of CCC.

scholarly journals Coordinate modulation of Na-K-2Cl cotransport and K-Cl cotransport by cell volume and chloride

2002 ◽  
Vol 283 (5) ◽  
pp. C1422-C1431 ◽  
Author(s):  
Christian Lytle ◽  
Thomas McManus
Keyword(s):  
Cell Volume ◽  
Cell Volume Regulation ◽  
Specific Effect ◽  
Feedback System ◽  
Major Effect ◽  
Cell Swelling ◽  
Cell Shrinkage ◽  
Set Point

Na-K-2Cl cotransporter (NKCC) and K-Cl cotransporter (KCC) play key roles in cell volume regulation and epithelial Cl− transport. Reductions in either cell volume or cytosolic Cl− concentration ([Cl−]i) stimulate a corrective uptake of KCl and water via NKCC, whereas cell swelling triggers KCl loss via KCC. The dependence of these transporters on volume and [Cl−]i was evaluated in model duck red blood cells. Replacement of [Cl−]i with methanesulfonate elevated the volume set point at which NKCC activates and KCC inactivates. The set point was insensitive to cytosolic ionic strength. Reducing [Cl−]i at a constant driving force for inward NKCC and outward KCC caused the cells to adopt the new set point volume. Phosphopeptide maps of NKCC indicated that activation by cell shrinkage or low [Cl−]iis associated with phosphorylation of a similar constellation of Ser/Thr sites. Like shrinkage, reduction of [Cl−]i accelerated NKCC phosphorylation after abrupt inhibition of the deactivating phosphatase with calyculin A in vivo, whereas [Cl−] had no specific effect on dephosphorylation in vitro. Our results indicate that NKCC and KCC are reciprocally regulated by a negative feedback system dually modulated by cell volume and [Cl−]. The major effect of Cl− on NKCC is exerted through the volume-sensitive kinase that phosphorylates the transport protein.

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