Changes In Cell Water Content Research Articles

Relaxometric investigations addressing the determination of intracellular water lifetime: a novel tumour biomarker of general applicability

ABSTRACT1H Fast-Field Cycling NMR relaxometry is proposed as a powerful method to investigate tumour cell metabolism by measuring changes in cell water content and mobility across the cellular membrane. Measurements of intracellular water residence time in cultured cells were carried out by measuring T1 at fixed field (0.2 T) after the addition of a paramagnetic Gd complex (Prohance) at different concentrations in the external medium. Investigations on tumour cells (mammary adenocarcinoma TS/A) grown in normo- or hypoxic conditions or suspended in ‘hypo-osmotic’ solutions allowed us to demonstrate that both hypoxic and hypo-osmotic conditions cause a marked increase in water mobility as assessed by the elongation of T1. Conversely, the metabolic change caused by glutamine (an aminoacid essential for tumour growth) deprivation caused a water mobility decrease (shorter T1). These findings suggest that T1 measurements at low and variable magnetic field strengths, giving access to the assessment of intracellular water lifetime, can provide important information about tumour cell metabolism in real time and non-invasively.

The ongoing search for the molecular basis of plant osmosensing.

### Introduction Cell viability and metabolism depend on cytoplasmic water and solute content, and organisms have evolved mechanisms to sense changes in cell water content, solute concentrations, cell volume, and/or turgor. This Perspective addresses the response to osmotic challenge in land plants

Isovolumetric regulation in mammal cells: role of taurine.

The ability to regulate cell volume is an ancient conserved trait present in essentially all species through evolution. The maintenance of a constant cell volume is a homeostatic imperative in animal cells. Changes in cell water content affecting the concentration of intracellular messenger molecules impair the complex signaling network, crucial for cell functioning and intercellular communication. Although the renal homeostatic mechanisms exert a precise control of extracellular fluid osmolarity, this is challenged in a variety of pathological situations. The intracellular volume constancy is continuously compromised by the generation of local and transient osmotic microgradients, associated with nutrients uptake, secretion, cytoskeleton remodeling and transynaptic ionic gradients1.

Influence of initial respiratory status on the short-and long-term activity of the trout red blood cell ?-adrenergic Na+/H+ exchanger

The effects of ambient O2 partial pressure and CO2 partial pressure on the intensity of rainbow trout (Oncorhynchus mykiss) red blood cell β-adrenergic Na+/H+ exchange were investigated. This was accomplished in vitro by continuously monitoring whole blood extracellular pH, partial pressures of O2 and CO2 and by measuring red blood cell water content and Na+ concentration before and 30 min after the addition of a catecholamine mixture (final nominal concentrations: 250 nmol·l-1 adrenaline and 20 nmol·l-1 noradrenaline). The experiments were performed under six different initial conditions combining two ambient partial pressures of CO2 (1.50 and 6.75 torr) and three ambient partial pressures of O2 (15, 30 and 150 torr). The activation of red blood cell Na+/H+ exchange (as indicated by marked reductions of whole blood pH) was followed by transient reductions in blood partial pressures of CO2 and O2 (2 min) resulting from the shift of the CO2/HCO3 - equilibrium within the cell and the subsequent binding of O2 to the haemoglobin. The initial reduction in blood CO2 partial pressure was followed by a rise reflecting the titration of plasma HCO3 - by extruded H+. At low partial pressure of CO2 (1.50 torr) there was a pronounced stimulatory effect of hypoxia on the initial intensity of the extracellular acidification (5 min), whereas at high CO2 partial pressure (6.75 torr) hypoxia actually lowered the extent of the initial acidification. In all cases, Na+/H+ exchange activation was accompanied by increases in cell water content and red blood cell Na+ levles when measured 30 min after addition of catecholamines. Both hypercapnia and hypoxia increased the magnitude of these changes although the largest changes in cell water content and Na+ levels were observed under hypercapnic conditions. Thus, the long-term activity (as determined by measuring cell water and Na+ levels) of the Na+/H+ exchanger was enhanced both by hypercapnia and hypoxia regardless of the initial CO2 partial pressure. The initial activity (5 min), on the other hand, although stimulated by hypercapnia was influenced by hypoxia in opposing directions depending upon the initial CO2 partial pressure of the blood.

Model for the role of macromolecular crowding in regulation of cellular volume.

A simple model is proposed to account for large increases in transporter-mediated ion flux across cell membranes that are elicited by small fractional changes of cell volume. The model is based upon the concept that, as a result of large excluded volume effects in cytoplasm (macromolecular crowding), the tendency of soluble macromolecules to associate with membrane proteins is much more sensitive to changes in cell water content than expected on the basis of simple considerations of mass action. The model postulates that an ion transporter may exist in either an active dephosphorylated state or an inactive phosphorylated state and that the steady-state activity of the transporter reflects a balance between the rates of phosphatase-catalyzed activation and kinase-catalyzed inactivation. Cell swelling results in the inhibition of kinase relative to phosphatase activity, thereby increasing the steady-state concentration of the active form of the transporter. Calculated volume-dependent stimulation of ion flux is comparable to that observed experimentally.

Osmotic water permeability of Necturus gallbladder epithelium.

An electrophysiological technique that is sensitive to small changes in cell water content and has good temporal resolution was used to determine the hydraulic permeability (Lp) of Necturus gallbladder epithelium. The epithelial cells were loaded with the impermeant cation tetramethylammonium (TMA+) by transient exposure to the pore-forming ionophore nystatin in the presence of bathing solution TMA+. Upon removal of the nystatin a small amount of TMA+ is trapped within the cell. Changes in cell water content result in changes in intracellular TMA+ activity which are measured with intracellular ion-sensitive microelectrodes. We describe a method that allows us to determine the time course for the increase or decrease in the concentration of osmotic solute at the membrane surface, which allows for continuous monitoring of the difference in osmolality across the apical membrane. We also describe a new method for the determination of transepithelial hydraulic permeability (Ltp). Apical and basolateral membrane Lp's were assessed from the initial rates of change in cell water volume in response to anisosmotic mucosal or serosal bathing solutions, respectively. The corresponding values for apical and basolateral membrane Lp's were 0.66 x 10(-3) and 0.38 x 10(-3) cm/s.osmol/kg, respectively. This method underestimates the true Lp values because the nominal osmotic differences (delta II) cannot be imposed instantaneously, and because it is not possible to measure the true initial rate of volume change. A model was developed that allows for the simultaneous determination of both apical and basal membrane Lp's from a unilateral exposure to an anisosmotic bathing solution (mucosal). The estimates of apical and basal Lp with this method were 1.16 x 10(-3) and 0.84 x 10(-3) cm/s.osmol/kg, respectively. The values of Lp for the apical and basal cell membranes are sufficiently large that only a small (less than 3 mosmol/kg) transepithelial difference in osmolality is required to drive the observed rate of spontaneous fluid absorption by the gallbladder. Furthermore, comparison of membrane and transepithelial Lp's suggests that a large fraction of the transepithelial water flow is across the cells rather than across the tight junctions.

L-929 cells under hyperosmotic conditions

Changes in cell water content resulting from sorbitol addition to the environment of L-929 cells were evaluated gravimetrically using 14C-labeled polyethylene glycol as a probe of extracellular space. Reductions in cell water were proportional to sorbitol supplements up to 0.6 molal, above which no further measurable decrease occurred. No volume regulation occurred for at least 1 h but the percentage of cell water lost was quickly regained when physiological conditions were restored. The amount of cell water lost because of a given hyperosmotic exposure was found to exceed the loss of cell volume. That discrepancy could be the result of an overestimation of extracellular space and/or an underestimation of cell volume reduction as a result of in-folding of the cell surface. Na+ and K+ were also measured in cells of variable water content and volume: no significant change occurred in the amounts of these ions per cell, but large increases in total cell concentration resulted from hyperosmotic exposure. The sum of Na+ and K+ concentrations exceeds the total osmotic pressure of the medium indicating that an appreciable fraction of Na+ and K+ must be bound to fixed charges within the cells. The results are evaluated in the context of intracellular organization.

Na-pump activity in rat kidney cortex cells and its relationship with the cell volume

The present work was undertaken to evaluate whether changes in cell water content of rat kidney cortex cells can modulate the transport activity of the ouabain-insensitive Na pump as they modulate the ouabain-insensitive Na +-ATPase. It was found that there is a close relationship between the cell volume and activity of the Na pump, whereas Na,K-pump activity is not affected by variations in cell volume. When the cell water content is low, Na-pump activity (Na + transport and Na +-ATPase activity) is minimal. Increases in cell water content produce a concomitant increase in Na-pump activity.

Cell volume regulation in goldfish intestinal mucosa.

1. Ion and water content of goldfish intestinal mucosa, stripped free from muscular layers were measured under various incubation conditions. 2. Ouabain induces an increase in cation content that is electrically compensated for by chloride. The increase in solute content is accompanied by an increase in water content. 3. When extracellular chloride is partially replaced by sulphate, ouabain does induce cell shrinkage. 4. Anoxia induces a rapid increase in cell volume that is restored by oxygenation of the incubation solution. Ouabain prevents the restoration of volume. 5. It is concluded that the classical ouabain-sensitive Na/K pump participates in the maintenance of cellular volume. We suggest that the constancy in volume after ouabain poisoning as is reported for many tissues might be due to a low chloride conductance of its membranes. 6. Anisotonic media (range: 0.6-1.2 isotonicity), made by variation on mannitol concentration, induce changes in cell water content that deviates from the simplified van't Hoff equation by about 10%. No change in water content after the initial increase was found. 7. We conclude that goldfish enterocytes do not possess a mechanism for rapid volume readjustment.

Response of active transport of ions and spontaneous water flux to osmotic gradients in gastric mucosa.

The effects of symmetric changes of the mucosal and serosal bathing solution on cell water content, net ion flux, and net water movement were studied in the isolated frog gastric mucosa. Similar to transmucosal concentration gradients that induce water movement and changes in cell water content, symmetric osmolality changes of the bathing solutions also produce changes in these functional parameters. Thus, increments from 165 to 286 mosmol/kg water in the osmolality of both solutions reduce cell water content from 2.37 plus or minus 0.12 to 1.30 plus or minus 0.20 ml/g wt, the net ion flux (acid secretion plus short-circuit current) from 4.83 plus or minus 0.36 to 3.44 plus or minus 0.26 mueq/cm2 per h, and the net water flux from 10.6 plus or minus 1.1 to 2.4 plus or minus 1.2 mul/cm2 per h. These osmotically induced flux changes of water and ions must be considered when osmotic gradients are being used to generate and to evaluate water movement across the gastric mucosa.

Micromanipulation of stomatal guard cells

THE opening and closing of stomatal pores is controlled by deformations of the guard cells resulting from changes in the pressure relationships between epidermal and guard cells. Under natural conditions such changes are, in turn, controlled by changes in the osmotic concentrations of cell saps or by changes in cell water content, although they could also be controlled by changes in cell wall properties. We have measured the pressures required to bring about or maintain stomatal openings by applying pressures directly within subsidiary cells and guard cells of Tradescantia virginiana and Vicia faba leaves.

Independence of water content and respiration in rabbit erythrocytes.

SummaryAccelerated O2 uptake of rabbit erythrocytes was measured in the presence of methylene blue and glucose during changes in erythrocyte water content. Although dependent upon substrate concentration, temperature, and pH, O2 uptake was independent of rather large changes in cell water content.