Cation effects on volume and water permeability in the halophilic algae Dunaliella parva.

Rates of change of cell volume were measured in suspensions of the halophilic green algae Dunaliella parva subjected to changes in cation composition and concentration of the outside medium. Measurements were made with a particle size analyzer and results were checked by direct microphotography. For any one salt solution, changes in cell volume with concentration were consistent with the Boyle-Van't Hoff model of an osmometer. Nonosmotic volume comprised 60-80% of total cell volume and was sensitive to the nature of the cation, increasing in the order Cs less than K less than Na less than Ca less than Mg. Kinetics of volume change in response to changes in outside salt concentration are best described by two kinetic coefficients differing by one order of magnitude and dependent on the nature of the outside cation (decreasing in order Cs greater than K greater than Na greater than Mg) as well as on direction of water flow.     

Electrical sizing of cells of the halophilic alga Dunaliella parva

Abstract. The cell volume of two strains of the halophilic alga Dunaliella parva was measured by two direct methods and by electrical sizing. The results from the direct methods agreed with each other and were consistently larger than those obtained from electrical sizing. Statistical analysis showed that the cell volume populations obtained by electrical size of D. Parva cells is due to the cell membrane having an unusually low electrical resistance relative to other cell membranes.     

The volume dependence of the erythrocyte water diffusion permeability

The product of the water diffusion permeability and the membrane area of a human erythrocyte has been found to be nearly independent of the cell volume. The product was measured by an NMR technique. This result conflicts with previous flow tube determinations but is in accord with recent measurements of the hydraulic permeability and various solute permeabilities. The results are consistent with the major part of the water fllux traversing the membrane through a fixed number of pores. There may also be a minor non-pore flux. It appears to be practicable to follow volume changes in the red blood cell by an NMR technique.     

Gallbladder epithelial cell hydraulic water permeability and volume regulation

The hydraulic water permeability (Lp) of the cell membranes of Necturus gallbladder epithelial cells was estimated from the rate of change of cell volume after a change in the osmolality of the bathing solution. Cell volume was calculated from computer reconstruction of light microscopic images of epithelial cells obtained by the "optical slice" technique. The tissue was mounted in a miniature Ussing chamber designed to achieve optimal optical properties, rapid bath exchange, and negligible unstirred layer thickness. The control solution contained only 80% of the normal NaCl concentration, the remainder of the osmolality was made up by mannitol, a condition that did not significantly decrease the fluid absorption rate in gallbladder sac preparations. The osmotic gradient ranged from 11.5 to 41 mosmol and was achieved by the addition or removal of mannitol from the perfusion solutions. The Lp of the apical membrane of the cell was 1.0 X 10(-3) cm/s . osmol (Posm = 0.055 cm/s) and that of the basolateral membrane was 2.2 X 10(-3) cm/s . osmol (Posm = 0.12 cm/s). These values were sufficiently high so that normal fluid absorption by Necturus gallbladder could be accomplished by a 2.4-mosmol solute gradient across the apical membrane and a 1.1-mosmol gradient across the basolateral membrane. After the initial cell shrinkage or swelling resulting from the anisotonic mucosal or serosal medium, cell volume returned rapidly toward the control value despite the fact that one bathing solution remained anisotonic. This volume regulatory response was not influenced by serosal ouabain or reduction of bath NaCl concentration to 10 mM. Complete removal of mucosal perfusate NaCl abolished volume regulation after cell shrinkage. Estimates were also made of the reflection coefficient for NaCl and urea at the apical cell membrane and of the velocity of water flow across the cytoplasm.     

Osmotic water permeability and regulatory volume decrease of rat thymocytes

Rat thymocytes displayed robust regulatory volume decrease (RVD) when suspended in NaCl-based hypotonic Ringer solutions. The RVD of thymocytes was completely abolished upon replacement of external Na+ ions with K+, indicating a role of coupled efflux of K+ and Cl- ions as a driving force of regulatory volume decrease. Osmotic water permeability (Pf) measured in KCl-based hypotonic solutions was (1.3 +/- 1.0 x 10(-4) cm/s at 25 degrees C and was temperature-dependent with low activation energy (Ea = 4.65 +/- 0.77 kcal/mol) characteristic to water transport through pores. HgCl2 and a sulfhydryl-blocking reagent, methyl methanethiosulphonate (MMTS), modulated the water permeability of thymocytes in a biphasic manner: inhibited at low dose (0.1-1 micromol/l) and restored or even enhanced at higher (10-100 micromol/l) concentrations. RVD paralleled the Pf: it was greatly suppressed at low dose of MMTS (sufficient to attenuate the water transport), but recovered at higher dose, when the water movement was restored. Therefore we suggest that thymocytes require the effective water transport for functional regulatory volume decrease.     

The antitheilerial effects of Theileria parva parva reaction and recovery sera in vitro

Peripheral blood leucocytes (PBL) of cattle were infected in vitro with the sporozoites of Theileria parva spp. The transformed cell lines were adapted to grow in sera from the PBL donors. The cattle were then infected with T. p. parva stabilate and either treated with parvaquone or the disease allowed to run its course. Sera harvested during severe disease reaction or early recovery were substituted for pre infection sera and caused the intracellular degeneration of the Theileria macroschizonts. Cell lines passaged in these sera died out as the parasites were eliminated. The antiparasitic effects of sera were short lived and were neither host nor parasite isolate restricted.     

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).     

氟西汀与三氯生复合暴露对麦穗鱼的毒性效应

为探讨药品和个人护理产品(PPCPs)复合污染对水生生态系统的影响,本试验以东北地区土著鱼类麦穗鱼为试验材料,研究了氟西汀(FLX)与三氯生(TCS)复合暴露对其不同器官的毒性效应.经急性(4 h)与慢性(42 d)复合暴露试验后分别检测麦穗鱼Ⅰ相和Ⅱ相解毒酶、神经系统、消化系统及抗氧化系统等受到的影响.结果表明: 在FLX/TCS复合暴露条件下,麦穗鱼脑部乙酰胆碱酯酶活性受到短暂抑制,肝中细胞色素P450活性持续受到抑制,肠中α-葡萄糖苷酶活性在急性暴露后受到诱导但是长期暴露后被抑制,同时长期复合暴露导致肝中脂质过氧化水平升高.氟西汀和三氯生对麦穗鱼的复合暴露可对麦穗鱼多个器官产生急性毒性应激效应,而随着暴露时间的延长,麦穗鱼可产生一定的适应性,但这种适应作用的机制有待进一步研究.     

Cation permeability induced by spermine and polybrene in rat liver mitochondria

The effect of the polybasic substances Polybrene and spermine on the passive and active transport of monovalent cations in mitochondria was studied. These agents were found to stimulate the low amplitude swelling of mitochondria. Volume oscillations were induced by addition of substrate in the presence of spermine. In conditions where weak oscillations were obtained without these substances, oscillations were stimulated and their frequencies increased in the presence of Polybrene and spermine. Their effects were maximal with 100–300 moles spermine per litre and 3–5 mg Polybrene per litre. These results are discussed in relation to an interaction of the agents studied with membrane negative charges which may be important regulators of ion transport.     

Cation specificity of propranolol-induced changes in RBC membrane permeability: comparative effects in human, dog and cat erythrocytes

Propranolol, in the presence of calcium, causes marked K efflux from human red blood cells (high K, low Na). The studies reported here indicate this effect of propranolol is specific for K and does not represent a nonspecific permeability increase for intracellular cations to leave the cell. Amphotericin-treated human RBC's (high Na, low K) and dog RBC's (high Na, low K) both gain K and increase in size when incubated in a K-medium containing propranolol and calcium. No effect was noted when cat RBC's (high Na, low K) were similarly treated. Propranolol, independent of added calcium, also inhibited the normally increased Na efflux observed when dog RBC's are suspended in K-medium. These species differences in response to propranolol thus may serve as a focus for elucidating the mechanism by which this drug alters normal membrane physiology. The unique drug effect on Na permeability of canine erythrocytes also may be a useful probe for the study of dog RBC volume regulation.     

Cation transport and cell volume changes in maturing rat reticulocytes

During maturation, reticulocytes lose membrane material,including transporters, and this is accompanied by a loss of cell waterand volume. Here we determined a possible role of ion transport inadjusting cell volume during maturation. Reticulocytes and red bloodcells of different ages were prepared from erythropoietin-treated ratsby density gradient fractionation. Cell volume and ion transport weremeasured in freshly prepared cells and in reticulocytes during in vitromaturation. Reticulocytes had an increased K content and cell volume,whereas intracellular Na was decreased. All parameters approached wholeblood values after 2 days in culture. Na-K pump was elevated inreticulocytes and decreased during maturation. Na-K-2Cl cotransport(NKCC) activity was lower in reticulocytes and was activated 8- and20-fold by shrinkage and okadaic acid, respectively, whereasstimulation was barely detectable in high-buoyant density red bloodcells. The ouabain- and bumetanide-insensitive Na flux in reticulocytesdecreased on maturation. Most of it was inhibited by amiloride,indicating the presence of Na/proton exchange. Our results show that,although the Na-K-pump activity in reticulocytes is very muchincreased, the enhanced capacity of NKCC is essentially cryptic untilstimulated. Both types of capacities (activities) decrease duringmaturation, indicating a possible loss of transport protein. Thedecrease was constrained to the period of reticulocyte maturation. Lossof transport capacity appears to exceed the loss of membrane surfacearea. Reticulocyte age-related changes in the net electrochemicaldriving force indicate that the increasing NKCC activity mightcontribute to the reduction in cell water.

     

Combined effects of light and water stress on chloroplast volume regulation.

A nuclear magnetic resonance technique was used to measure changes in the water content of Acer platanoides chloroplasts in leaf discs that had reached osmotic equilibrium with external solutions either in the dark or under exposure to light. Results showed that chloroplast volume regulation (CVR) maintained constant water content in the chloroplasts over a range of water potentials in the dark, but CVR failed when the water potential fell below a critical value. The critical potential was lower in the dark in sun leaves than in shade leaves. Upon exposure to intense light, CVR remained effective in sun leaves over the same range as in the dark, but it failed in shade leaves at all water potentials. Osmolytes are necessary for CVR, but KCl is relatively ineffective; increased concentrations of intracellular KCl did not fully support an increase in the range of CVR. The results indicate that leaves need reserve supplies of cytosolic osmolytes to maintain CVR at low water potentials, and a larger reserve supply is needed in leaves that are exposed to intense light.     

Cell volume and plasma membrane osmotic water permeability in epithelial cell layers measured by interferometry.

The development of strategies to measure plasma membrane osmotic water permeability (Pf) in epithelial cells has been motivated by the identification of a family of molecular water channels. A general approach utilizing interferometry to measure cell shape and volume was developed and applied to measure Pf in cell layers. The method is based on the cell volume dependence of optical path length (OPL) for a light beam passing through the cell. The small changes in OPL were measured by interferometry. A mathematical model was developed to relate the interference signal to cell volume changes for cells of arbitrary shape and size. To validate the model, a Mach-Zehnder interference microscope was used to image OPL in an Madin Darby Canine Kidney (MDCK) cell layer and to reconstruct the three-dimensional cell shape (OPL resolution < lambda/25). As predicted by the model, a doubling of cell volume resulted in a change in OPL that was proportional to the difference in refractive indices between water and the extracellular medium. The time course of relative cell volume in response to an osmotic gradient was computed from serial interference images. To measure cell volume without microscopy and image analysis, a Mach-Zehnder interferometer was constructed in which one of two interfering laser beams passed through a flow chamber containing the cell layer. The interference signal in response to an osmotic gradient was analyzed to quantify the time course of relative cell volume. The calculated MDCK cell plasma membrane Pf of 6.1 x 10(-4) cm/s at 24 degrees C agreed with that obtained by interference microscopy and by a total internal reflection fluorescence method. Interferometry was also applied to measure the apical plasma membrane water permeability of intact toad urinary bladder; Pf increased fivefold after forskolin stimulation to 0.04 cm/s at 23 degrees C. These results establish and validate the application of interferometry to quantify cell volume and osmotic water permeability in cell layers.