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PubMed1983

Osmotic water permeability of the human red cell. Dependence on direction of water flow and cell volume.

The osmotic permeability coefficient (Pf) was measured with a stopped-flow light-scattering technique. There is an artifactual light-scattering signal produced by the initial mixing that decays with a half-time of approximately 0.2 s. This seriously interferes with the measurement of the osmotically induced change in cell volume, which has a similar half-time. This "injection artifact" is associated with the biconcave shape of the cells. It is negligible for cells that have been made nearly spherical by swelling them in 160 mosmol. The dependence of this artifact on the cell volume may explain the previously observed dependence of Pf on the cell volume. When cells are made echinocytic (and therefore spherically symmetric), this injection artifact becomes negligible at all cell volumes and Pf can be accurately measured. The Pf of echinocytic cells was nearly constant, varying by less than 10% with the direction of flow and the medium osmolarity (160-360 mosmol). The average value of Pf was 2.0 X 10(-2) cm/s (T = 23 degrees C).

Cell Membrane Permeability↗
PubMedDate not supplied

Osmotic balance in the eggs of the turtle Chelodina rugosa during developmental arrest under water.

The tropical Australian turtle Chelodina rugosa normally lays its hard-shelled eggs in mud, under shallow freshwater, during the monsoon season. The eggs undergo developmental arrest until the water recedes and oxygen is able to diffuse into the embryo. This period of arrest can exceed 12 wk without embryonic mortality. To understand how the eggs avoid osmotic absorption of water leading to shell rupture and embryonic death, this study investigates the solute concentrations and volumes of the albumen and yolk compartments during submergence in distilled water. The albumen loses considerable sodium through the shell, particularly during the first week, and its osmotic concentration drops from 234 mmol/kg at laying to about 23 mmol/kg. Meanwhile, water from the albumen slowly moves through the vitelline membrane into the yolk compartment, which enlarges at a constant rate until it approaches the inside of the shell at about 22 wk. Osmotic uptake dilutes yolk solutes, decreasing the osmotic concentration from 281 mmol/kg at laying to 132 mmol/kg at 157 d. Loss of embryonic viability is associated with contact of the vitelline membrane with the inside of the shell. The principal adaptation of this species for protracted developmental arrest under water is a vitelline membrane of such low permeability to water that the expansion of the yolk compartment occurs about 10 times more slowly than in other chelonians.

Animals↗
PubMed1966

The effect of membrane fixed charges on diffusion potentials and streaming potentials.

1. Electrical potential differences (p.d.'s) have been measured across an in vitro preparation of rabbit gall-bladder.2. When the gall-bladder separates identical bathing solutions, the p.d. is always zero, regardless of the composition of the bathing solution. Hence the gall-bladder is symmetrical: i.e. the mucosal and serosal cell membranes have the same relative permeability coefficients.3. Osmotic water flow causes streaming potentials of up to 20 mV, of a sign indicating greater permeability to cations than to anions.4. At constant osmolarity, streaming potentials increase slightly with NaCl concentration. Streaming potentials decrease considerably with changes in osmolarity resulting from changes in NaCl concentration.5. Diffusion potentials resulting from electrolyte concentration gradients are fitted well by the constant-field equation with the relative permeability coefficients P(Na) = 1.00, P(Cl) = 0.33, P(K) = 2.3. These permeability coefficients are independent of osmolarity and of salt concentration.6. Relative to 0.25 mM-Ca, 5 mM-Ca reduces streaming potentials by 40%, NaCl diffusion potentials by 62%, and potassium diffusion potentials by 43%.7. The aqueous channels through which water and electrolytes traverse the cell membranes of the gall-bladder contain negative fixed charges, which are blocked by Ca. The physiological significance of the charges may be to reduce chloride permeability and thereby to increase the effectiveness of the gall-bladder in concentrating bile.8. The effect of pH, and analogy with surface charges of other cells, suggest that the charges are organic acids of low pK(a).

Animals↗
PubMed1966

A rapid method for determining voltage-concentration relations across membranes.

1. A rapid method is described for obtaining curves relating the potential difference (p.d.) across a membrane to solute concentration gradients. The experiment consists of measuring the p.d. as a function of time while the concentration at the membrane is continuously increasing from 0 to C(o) (or decreasing from C(o) to 0) owing to diffusion through an unstirred layer.2. The voltage trace is then analysed with a piece of transparent paper, on which lines corresponding to solutions of the diffusion equation convert the time axis of the voltage trace into a concentration axis. Superposition of this paper on an experimental voltage trace permits one to read off the p.d. corresponding to any concentration between 0 and C(o).3. The relation between diffusion potentials and NaCl concentration, and between streaming potentials and concentrations of impermeant non-electrolytes, has been reconstructed by this procedure in rabbit gall-bladder. The method yields non-linear voltage-concentration curves in good agreement with those obtained by conventional methods, but requires much less time and experimental manipulation. The conditions that must exist for the method to be valid are discussed.4. The thicknesses of the unstirred layers in rabbit gall-bladder are measured, and are shown to have a negligible effect upon steady-state streaming potentials and NaCl diffusion potentials.5. The changes of permeability with osmolarity described in rabbit gall-bladder must be complete within a fraction of a second, in agreement with the interpretation that they represent swelling and shrinking of the cell membrane itself.

Animals↗
PubMed1976

Osmotic reflextion coefficients of capillary walls to low molecular weight hydrophilic solutes measured in single perfused capillaries of the frog mesentery.

1. Individual capillaries of the transilluminated frog mesentery have been perfused with suspensions of human red cells in frog Ringer solution containing 1-0 g albumin 100 ml.-1. The outer surface of the mesentery has been washed with normal frog Ringer solution and with frog Ringer solutions made hypertonic by addition of one of the following solutes: sodium chloride (100 m-mole. 1.-1); urea (100 m-mole.1.-1); sucrose (20-50 m-mole. 1.-1); cyanocobalamin (8-5 m-mole. 1.-1). The temperature of the mesentery was between 14 and 16 degrees C in all experiments. 2. Wtih the mesentery superfused with normal Ringer, the filtration coefficient was determined from measurements of the rate of fluid filtration across the capillary wall, at a series of known capillary pressures (Michel, Mason, Curry & Tooke, 1974). Filtration coefficient varied from 0-69 X 10(-3) to 4-45 X 10(-3) mum. sec-1 .cm H2O-1 with an average value of 1-87 X 10(-3) mum. sec-1. cm H2O-1. 3. When the superfusate was made hypertonic by the addition of a test solute, the osmotic reflextion coefficient (sigma) of the capillary wall to test solute was calculated from the additional rate of filtration, the concentration of test solute in the superfusate and the filtration coefficient. Average values for sigma were: sodium chloride, 0-068 +/- 0-03 (three capillaries); urea, 0-071 +/- 0.015 (four capillaries); sucrose, 0-115 +/- 0-023 (seven capillaries); cyanocobalamin, 0-100 +/- 0-03 (three capillaries). 4. In further experiments, the osmotic reflextion coefficients to sodium chloride, urea and sucrose were determined in the same capillary. Five technically acceptable experiments were carried out. Although there were differences in the value of sigma between different capillaries, in any one capillary values of sigma were of the same magnitude and there appeared to be no significant trend with the molecular size of the test solute. 5. Our findings are inconsistent with the hypothesis that there is a single pathway for water and small hydrophilic molecules across the capillary wall. 6. Our results may be interpreted in terms of an exclusive channel for water in parallel with a channel shared by both water and small hydrophilic molecules. It is suggested that the exclusive water channel may be the membranes and cytoplasm of the endothelial cells and the shared channel may be located in the intercellular junctions. 7. Our data suggest the exclusive water channel represents about 10% of the total filtration coefficient in frog mesenteric capillaries. The shared channel shows relatively little restriction to the molecules investigated. Estimates of the volume flow throught the two channels are made for conditions where hydrostatic pressure differences and osmotic pressure differences are the driving forces.

Animals↗
PubMed1976

Permeability of individual human erythrocytes to thiourea.

The osmotic swelling to haemolysis of individual red blood cells by isosmotic thiourea has been studied using microcine photography. 2. Crenation occurs immediately upon addition of isosmotic thiourea. The cell becomes a crenated sphere without volume decrease. 3. Subsequently, the cell volume increases linearly with time with maximum swelling occurring at about 102 sec which is 81% of the total haemolysis time. 4. At maximum swelling, the cell volume is 92% greater than the initial cell volume. This volume increase is about double that measured with other permeating substances. 5. The much larger maximum volume implies that thiourea increases the area of the cell membrane. This increase varies from 0 to 75% for individual cells, with a mean of 22%. 6. Membrane expansion varies inversely as the initial cell membrane area and cell volume (r=0-790). 7. Using the increased surface area, increased maximum volume and the swelling time, the mean permeability is calculated to be 5-52 X 10(-7) cm/sec (S.D. of mean=+/-1-19 X 10(-7) cm/sec). The distribution of permeabilities represents a normal distribution. 8. The pre-lytic potassium loss ranged from 0 to 36% with a mean value of 16-5%. This is consistent with values reported in the literature for slow haemolysis. As with other permeants the distribution is skewed towards lower values. 9. Membrane permeability of individual cells varies with the amount of membrane expansion observed. Coefficient of correlation between permeability and expansion index is 0-674. 10. There is no correlation between permeability and the reciprocal of the haemolysis time (r=-0-035). The correlation between permeability and the reciprocal of the swelling time is also poor (r=0-303), probably owing to the variability in membrane expansion by thiourea in individual cells. 11. As has been shown previously for faster permeants, the permeability coefficient cannot be calculated from the haemolysis time. Because thiourea alters the membrane area and the haemolytic volume, the coefficient cannot be calculated from the swelling time unless the changes in the membrane area are also taken into account.

Cell Membrane↗
PubMed1988

Filtration coefficient and osmotic reflection coefficient to albumin in rabbit submandibular gland capillaries.

1. The isolated perfused submandibular salivary gland of the rabbit has been used in order to make estimates of the filtration coefficient (Kf) and reflection coefficient (sigma d) of the capillary wall to albumin. 2. An isogravimetric preparation was used and in paired experiments the value for Kf obtained in glands perfused with albumin-Krebs solution, 0.96 +/- 0.086 (mean +/- S.E. of mean) ml min-1 mmHg-1 100 g-1, was not significantly different from that in blood-perfused glands, 0.90 +/- 0.15. 3. On analysing the data for reflection coefficient, it was concluded that the above values underestimated Kf by about 30%; using corrected values for Kf, osmotic reflection coefficients were determined from the weight changes following a sudden change in the oncotic pressure of the perfusate. The value for sigma d to albumin lay between 0.79 and 1.0, the lower value being obtained after the Kf correction. 4. The high hydraulic conductivity, combined with sieving properties comparable to those in continuous capillaries, is consistent with other data on fenestrated capillaries. 5. Finally, it was observed that, while the Kf value calculated from the initial flux rate was similar whether measured during fluid efflux from or influx into the microvasculature, on returning to the initial conditions after raising osmotic pressure, efflux was now more rapid than influx. This phenomenon is discussed in relation to readjustment of Starling forces and the possible existence of an asymmetric double membrane in the capillary, interstitium system and cells of the salivary gland.

Albumins↗
PubMed1984

Effect of an osmotic gradient on antidiuretic hormone-induced endocytosis and hydroosmosis in the toad urinary bladder.

The luminal (apical) border of the epithelium of the bladder in the well-hydrated toad is relatively impermeable, so the bladder usually stores hyposmotic urine. When antidiuretic hormone (ADH) increases apical membrane osmotic permeability dramatically, water is resorbed from hyposmotic mucosal solution; in the presence of hyposmotic or isosmotic mucosal solutions, ADH concomitantly induces exocytosis at the apical border of granule-rich (G) cells. Then ADH induces endocytosis at this border. We describe how an osmotic gradient affects ADH-induced endocytosis and hydroosmosis in vitro. We can assess ADH-induced endocytosis in gradient and no-gradient bladders by applying a double-marker technique that distinguishes among endocytosis, completed internalization of previously surface-attached membrane, and surface invagination by comparing the number of horseradish peroxidase (HRP) uptake bodies (endocytosis) with the number of ruthenium red (RR)-delineated bodies (surface invaginations). With this approach we find that gradient bladders have approximately six times more ADH-induced endocytosis than no-gradient bladders during 45-60 min of ADH stimulation. Furthermore, at 60 min approximately 50% of the HRP-containing structures in no-gradient bladders remain surface connected compared with approximately 1% in gradient bladders. In parallel physiological studies, no-gradient bladders reach and maintain higher induced osmotic permeabilities than gradient bladders. These findings support the hypothesis that endocytosis plays an active role in reestablishing impermeable apical surface characteristics in toad bladder.

Animals↗
PubMed1985

Blockade of colchicine-induced inhibition of vasopressin-stimulated osmotic water flow: failure to influence microtubule formation.

Colchicine inhibits vasopressin-induced osmotic water flow across isolated toad urinary bladder. Concomitantly, colchicine has been shown to reduce the relative cytoplasmic volume fraction of microtubules in the apical granular cells of this epithelium that have been shown previously to mediate the hydroosmotic effect of vasopressin. Therefore, an intact cytoskeleton has been postulated to be a requirement for a full response to vasopressin. Since it has been demonstrated recently that cyclooxygenase inhibitors (meclofenamic acid) abrogate the inhibition by colchicine of vasopressin-stimulated water flow, we tested by stereological criteria the hypothesis that colchicine in the presence of meclofenamic acid does not prevent the polymerization of tubulin. Our results show that the relative cytoplasmic volume fraction of microtubules was reduced 75% by colchicine in the presence or absence of meclofenamic acid. An alternative explanation of the inhibitory action of colchicine is its ability in the toad urinary bladder to enhance the endogenous synthesis of and sensitivity to prostaglandin E, a potent negative modulator of vasopressin-stimulated water flow. An intact microtubular component of the cytoskeleton does not appear to be required for a maximal response to a physiological dose of vasopressin.

Animals↗
PubMed1987

Comparison of effects of forskolin, cAMP, and vasopressin on Pf/Pd(w) of toad urinary bladder luminal membrane.

We have reported that Pf/Pd(w) (the ratio of osmotic and diffusional water permeabilities) for the luminal membrane of toad urinary bladder is approximately 17 for tissues stimulated with either vasopressin or 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). In a recent abstract, Kachadorian and co-workers have shown that tissues stimulated with adenosine 3',5'-cyclic monophosphate (cAMP) or forskolin have a lower Pf than would be anticipated from the frequency of aggregates visualized on the flat portion of the luminal membrane using freeze-fracture electron microscopy. We report here measurements of Pf/Pd(w) for the luminal membrane of tissues receiving these agents: Pf/Pd(w) for submaximally stimulated tissues was the same, regardless of whether the stimulant was vasopressin (12.7 +/- 0.3), forskolin (13.7 +/- 0.9), or cAMP (12.0 +/- 1.3). The calculated Pd(w)'s for the series barrier were also identical (6.8 +/- 0.5, 6.5 +/- 0.3, and 8.2 +/- 1.0 X 10(-4) cm/s respectively). Our data, taken together with those of Kachadorian et al. are consistent with a number of possibilities: because our methodology does not permit estimation of Pf for the series barrier, we cannot rule out the possibility of a "post-luminal barrier" that is rate-limiting for Pf, but not for Pd(w) in forskolin- and cAMP-stimulated tissues, Pf and Pd(w) of the luminal surface aggregates could decrease in parallel, so that luminal membrane Pf/Pd(w) remains constant, and there could be a diminished frequency of fused aggregate-rich aggrephores, but not of aggregates that are on the flat portion of the luminal membrane. Only the latter can be unequivocally quantitated using freeze-fracture electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1967

The application of Starling's law of capillary exchange to the lungs.

The forces governing the movement of water across the pulmonary capillaries were studied in 39 intact, spontaneously breathing dogs. A situation favoring the net movement of water out of the pulmonary capillaries was created by means of partial pulmonary venous obstruction (left atrial balloon catheter) followed by rapid saline hemodilution. A predetermined difference between pulmonary capillary and plasma colloid osmotic pressures was maintained for periods of 1 to 2 hours. Left atrial (P(LA)) and plasma colloid osmotic pressures (pi(pl)) were measured directly. The water content of the lungs was measured serially by an indicator-dilution technique, and at autopsy by drying the lungs. The rate of accumulation of lung water was measured in four groups of animals: in three of the groups, the capillary hydrostatic and colloid osmotic pressures were varied; in the fourth group, the right lymphatic duct was obstructed in addition. The average rate of water accumulation in the lungs varied in a nonlinear way with the level of the capillary hydrostatic-plasma colloid osmotic pressure difference and was unaffected by the level of the capillary hydrostatic pressure. At low levels of P(LA) - pi(pl), water accumulated in the lung at an average rate of 0.09 g per g dry lung per hour per mm Hg pressure difference. At higher levels of P(LA) - pi(pl) the average rate of accumulation was 0.22 g per g per hour per mm Hg DeltaP; in most of the experiments in this group water accumulated in the lungs slowly during the first 30 minutes of the test period and more rapidly as the period was extended. Obstruction of right lymphatic duct outflow did not alter the rate of water accumulation. Based on the control data of the present experiments, the pericapillary pressure in normal lungs is estimated to be of the order of - 9 mm Hg in the normal dog lung. The filtration coefficient for the pulmonary capillaries is estimated to be of the order of one-tenth to one-twentieth of that for canine muscle capillaries. The data of the present study indicate that edema formation in lung tissue cannot be defined solely in terms of intravascular forces, but may be governed to a significant degree by changes in pericapillary forces in the pulmonary interstitium.

Animals↗