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PubMed1995

Molecular mechanisms for passive and active transport of water.

Water crosses cell membranes by passive transport and by secondary active cotransport along with ions. While the first concept is well established, the second is new. The two modes of transport allow cellular H2O homeostasis to be viewed as a balance between H2O leaks and H2O pumps. Consequently, cells can be hyperosmolar relative to their surroundings during steady states. Under physiological conditions, cells from leaky epithelia may be hyperosmolar by roughly 5 mosm liter-1, under dilute conditions, hyperosmolarities up to 40 mosm liter-1 have been recorded. Most intracellular H2O is free to serve as solvent for small inorganic ions. The mechanism of transport across the membrane depends on how H2O interacts with the proteinaceous or lipoid pathways. Osmotic transport of H2O through specific H2O channels such as CHIP 28 is hydraulic if the pore is impermeable to the solute and diffusive if the pore is permeable. Cotransport of ions and H2O can be a result of conformational changes in proteins, which in addition to ion transport also translocate H2O bound to or occlude in the protein. A cellular model of a leaky epithelium based on H2O leaks and H2O pumps quantitatively predicts a number of so-far unexplained observations of H2O transport.

Biological Transport, Active↗
PubMed1998

Transdermal permeation of neutral molecules by skin electroporation.

Electroporation of skin has recently been shown to enhance transport of charged molecules across skin by up to four orders of magnitude. This study demonstrates that high-voltage pulses can also increase transdermal permeation of two neutral model solutes, i.e. mannitol and water, up to 100-fold. The elevated flux results from the persistent increase in skin permeability following electroporation, rather than from electro-osmosis during pulsing. Control on transport was achieved by controlling the electrical parameters of the pulse, i.e. the pulse voltage, time constant and number.

Animals↗
PubMed1990

Hydraulically-induced convective solute transport across the rabbit peritoneum.

Transport of solutes during osmotically-induced transperitoneal ultrafiltration is less than would be predicted based upon rates of transperitoneal solute diffusion. Previous workers have hypothesized that osmotically-induced convective solute transport occurs only in small pores at the arteriolar end of peritoneal capillaries, whereas solute diffusion occurs only through large venular pores. We tested this heteroporosity hypothesis in the eviscerated New Zealand White rabbit by determining sieving coefficients (S) for creatinine, p-aminohippurate (PAH) and neutral dextran during hydraulically-induced transperitoneal ultrafiltration (N = 13). A hydraulically-induced driving force directs convective solute transport through the same capillary pores employed for diffusion; therefore S for all solutes should approach unity if the heteroporosity hypothesis is valid. S for creatinine and PAH were respectively 0.72 +/- 0.03 and 0.67 +/- 0.05, values lower than unity and not different from those previously determined during osmotically induced ultrafiltration. Mean S for dextran were relatively independent of molecular size, ranging from 0.50 at 13 A to 0.40 at 50 A. Thus, dextran S were higher than those previously determined during osmotically induced ultrafiltration yet still less than unity. Control experiments (N = 6) suggested that only surface area and not transport characteristics were altered by evisceration. These observations demonstrate that the heteroporosity hypothesis fails to completely describe both diffusive and convective transport properties of the peritoneum.

Animals↗
PubMed1973

A sensitive and accurate gel osmometer.

1. A bilayer strip, cut from a thin layer of cross-linked polyacrylamide gel cast on to cellulose tissue, forms an open circular loop whose ends are close together. Shrinkage of the gel, in response to the osmotic pressure of a non-penetrating solution, causes a proportional separation of the ends of the loop. This is measured with a microscope and micrometer eyepiece. 2. The resulting effective sensitivity is about 30 times that of the Sephadex-bead osmometer (Ogston & Wells, 1970), i.e. of the order of 5Pa, comparable with that of a membrane osmometer. Use of gel up to 70% (w/v) allows the measurement of molecular weights, as low as 1500 in favourable cases, with an accuracy of 1-2%. The useful range of osmotic pressure is up to 5kPa. A single measurement requires 0.5ml of solution. Equilibration is completed in 20-30min. 3. The method is illustrated by measurements on human serum albumin, ovalbumin, cytochrome c, samples of dextrans, polyvinyl alcohol, and polyethylene glycols 6000 and 1000.

Acrylamides↗
PubMed1960

Osmotic flow of water across permeable cellulose membranes.

Direct measurements have been made of the net volume flow through cellulose membranes, due to a difference in concentration of solute across the membrane. The aqueous solutions used included solutes ranging in size from deuterated water to bovine serum albumin. For the semipermeable membrane (impermeable to the solute) the volume flow produced by the osmotic gradient is equal to the flow produced by the hydrostatic pressure RT DeltaC, as given by the van't Hoff relationship. In the case in which the membrane is permeable to the solute, the net volume flow is reduced, as predicted by the theory of Staverman, based on the thermodynamics of the steady state. A means of establishing the amount of this reduction is given, depending on the size of the solute molecule and the effective pore radius of the membrane. With the help of these results, a hypothetical biological membrane moving water by osmotic and hydrostatic pressure gradients is discussed.

Cell Membrane↗
PubMed2000

Water permeability and revolving movement in Phaseolus vulgaris L. twining shoots.

Osmotic water permeability (Pos) was measured in protoplasts isolated from different tissues of Phaseolus vulgaris twining shoot. Parenchyma protoplasts exhibited more frequently high Pos values than epidermis protoplasts did. Water channels could facilitate water movement between parenchyma cells whereas cell-to-cell water transport mostly occurs through plasmodesmata in epidermis.

Cell Membrane Permeability↗
PubMed1986

Selectivity to K+ and Na+ of protoplast fractions isolated from different regions of Aspergillus nidulans hyphae.

The selectivity to K+ and Na+ of protoplast samples representing cytoplasm isolated from different regions of the hyphal filament of Aspergillus nidulans was investigated. Concentrations of both ions contained in successive protoplast fractions were measured. During lytic digestion, protoplasts were released first from apical regions and subsequently from progressively older regions of hyphae. A low K+/Na+ ratio was found in protoplasts containing primarily apical cytoplasm and a high K+/Na+ ratio was found in protoplasts originating from older regions of hyphae. The ratios were the same whether MgSO4 or mannitol was used as stabilizer. Absolute concentrations of both ions were higher in protoplasts of apical origin. Protoplasts stabilized in mannitol lost more ions than those stabilized in MgSO4 over an 8 h incubation period. Na+ losses were higher from apical protoplasts whereas K+ losses were higher from protoplasts liberated from older regions of hyphae. The addition of divalent metal cations (1.5 mM-Mn2+ or Mg2+) reduced losses of Na+ from protoplasts but did not affect loss of K+. Data obtained using protoplast samples were related to those obtained for intact mycelium. Absolute losses of both ions from mycelium were lower than for protoplasts but when compared on a protein basis the data suggested that protoplasts possess properties similar to those of intact mycelium in terms of K+ and Na+ selectivity.

Aspergillus nidulans↗
PubMed1993

Cation flux studies of the lesion induced in human erythrocyte membranes by the thermostable direct hemolysin of Vibrio parahaemolyticus.

Vibrio parahaemolyticus, an important agent of seafood-borne gastroenteritis, expresses several putative virulence factors that could account for the disease symptoms of infected humans, namely, diarrhea, nausea, and abdominal cramps. The pathogenicity of V. parahaemolyticus correlates well with the Kanagawa phenomenon (the hemolytic ability of strains grown on Wagatsuma blood agar), implicating the thermostable direct hemolysin (TDH) as the predominant toxin responsible for pathogenicity. TDH-induced hemolysis could be inhibited by the addition of the osmolyte sorbitol to the extracellular solution, supporting the hypothesis that hemolysis occurs through colloid osmosis secondary to an increase in the cation permeability of the membrane. The effect of TDH on cation permeability was investigated by measuring K+ (congener, 86Rb+) influx into human erythrocytes in which the endogenous cation transporters had been blocked (by use of ouabain, bumetanide, and nitrendipine). TDH increased K+ influx into these cells; this increase was rapid in onset and constant in magnitude, suggesting a direct action by TDH on the membrane. The kinetics of leak generation were examined; the relationship between counts accumulated and hematocrit indicated that the TDH-induced lesion is multihit in nature. TDH-induced K+ influx was sensitive to Zn2+. Time courses of hemolysis in isosmotic solutions of monovalent cation chlorides were used to obtain the selectivity series for the TDH-induced leak: Cs+ > Li+ > K+ > Rb+ > Na+. Both the Zn2+ sensitivity and this selectivity series were obtained for crude culture supernatants, suggesting that TDH is the predominant leak-inducing agent. Thus, we have identified several features of the TDH-induced leak likely to be important in the diarrhetic action of V. parahaemolyticus in the human intestine.

Adult↗
PubMed1996

Swelling of the vesicle is prerequisite for PTH secretion.

Unlike most secretory cells, high extra cellular calcium inhibits rather than stimulates hormonal secretion in several cells such as parathyroid cells, Juxtaglomerular cells and osteoclast. To gain further insight into the common but unique stimulus-secretion coupling mechanism in these cells, bovine parathyroid slices were incubated in various conditions of Krebs-Ringer (KR) solution containing essential amino acids. Parathyroid cells showed the inverse dependency of secretion on extra cellular calcium concentration as we expected. Ammonium acetate overcame the inhibitory effect of 2.5 mM of calcium and the maximum effect was as much as the five times of the basal value, while there was a little additive effect under 0 mM CaCl2. PTH secretion was biphasic according to the change of extra cellular osmolarity and the lowest response was observed at 300 mOsm/l. In Na-rich KR solution, high concentration of nigericin (> 10(-4)M) completely overcame the inhibitory effect of 2.5 mM CaCl2 and the maximum stimulatory effect was 8 times greater whereas it was only 2 times greater without CaCl2. In K-rich KR solution that abolished the K-gradient between the extra cellular solution and the cytoplasm, the rate of PTH secretion increased, and furthermore the addition of nigericin increased the rate of secretion significantly. The results above suggested that the osmotic swelling of the secretory vesicle in parathyroid cells might promote exocytosis as in Juxtaglomerular cells. We propose that the swelling of the vesicle is also prerequisite for secretion in several cells inhibited paradoxically by Ca++, whatever the signal transduction pathway for swelling of the secretory granules induced by the lowering of Ca++ in cytoplasm are.

Acetates↗
PubMed2004

Effects of acetazolamide and anordiol on osmotic water permeability in AQP1-cRNA injected Xenopus oocyte.

AIM: To study the effects of acetazolamide and anordiol on osmotic water permeability in aquaporin 1 (AQP1)-cRNA injected Xenopus oocyte and their mechanisms. METHODS: AQP1 gene constructed in pBluescript was transcripted into cRNA in vitro and then the cRNA was injected in Xenopus oocytes. The effects of acetazolamide and anordiol on the water transport function of AQP1 were observed by assaying the osmotic swelling of oocytes. In addition, their effects on protein expression of AQP1 were quantitatively investigated by Western blotting method. RESULTS: After incubation for 15 min or 72 h, acetazolamide, a carbonic anhydrase inhibitor, equally reduced the water permeability of AQP1-cRNA injected oocyte in a dose-dependent manner. After incubation for 72 h, anordiol, an antiestrogen with partial estrogenic activity, reduced the osmotic water permeability dose dependently as well; however, no discernable action was observed after incubation with anordiol for 15 min. The Western blotting analysis showed that acetazolamide did not influence the protein expression of AQP1. However, after incubation for 72 h with anordiol (10 micromol/L), the quantity of AQP1 in the oocyte membrane was decreased dramatically (P<0.05). CONCLUSION: Both acetazolamide and anordiol inhibited the osmotic water permeability of AQP1-cRNA injected oocyte, but their mechanisms were different. Acetazolamide functionally inhibited the osmotic water permeability of AQP1, whereas anordiol primarily decreased the amount of AQP1 protein in the oocyte membrane.

Acetazolamide↗
PubMed1988

On the role of calcium in the mechanism of ADH action.

With an increased influx of Ca2+ in the cytoplasm, the response of cells to ADH in the urinary bladder of the frog was lowered by addition of ionophore A23187 from the side of the basolateral cell membrane, but inhibited when it was added from the apical cell membrane. The removal of calcium by EGTA from the serosal surface was accompanied by a sharp increase of osmotic permeability not only to water, but also to inulin; while when calcium was removed from the mucosal surface of the urinary bladder, osmotic permeability was not changed. After being added to the Ringer solution from the outer surface of the apical cell membrane, the inhibitors of Ca2+ channels (verapamil, Ni2+, Mn2+, Co2+) decreased the effect of ADH. These data indicate that Ca2+ applied onto the outer surface of apical plasma membrane plays an important role in the action of ADH.

Animals↗