Osmotic pressure induced pores in phospholipid vesicles.

We report a comparative study of the leadage of hydrophilic molecules from vesicles of egg lecithin (EL) and of dipalmitoyllecithin (DPL). The effect of osmotic pressure differences a leakage is consistent with a model for statistical pore nucleation process. The major difference in osmotic pressure induced leakage from DPL and EL is that the number of pore creation sites is much greater in DPL. We suggest that the difference in number of these sites also accounts for other differences in the properties of DPL and EL, namely for differences in vesicle fusion and apparent rate of "flip-flop".     

渗透胁迫对皱果苋叶片渗透调节物质的影响

目的:研究渗透胁迫下皱果苋的渗透调节物质变化,探讨皱果苋的耐旱机理.方法:分别用浓度为5%、10%和20%PEG6000进行渗透胁迫,处 理皱果苋幼苗0、1、3、5和7d,取叶片测定各项生理指标.结果:随着渗 透胁迫时间的延长,皱果苋幼苗叶片中相对含水量(RWC)逐步下降;而脯氨酸和可溶性糖含 量上升;在5d,5%、10%、20%PEG胁迫下的叶片脯氨酸含量分别是对照的3.73、6.27、9.77 倍;而叶中可溶性糖含量分别是对照的2.87、3.68、5.21倍.胁迫3d,20%PEG处理的材料出 现萎焉现象,胁迫5d,10%PEG处理的材料开始萎焉;在5%和10%PEG胁迫下,可溶性蛋白含量 升高,但在20%PEG胁迫下,可溶性蛋白含量先升后降.结论:在适度的渗 透胁迫下,皱果苋叶片中的渗透调节物质使其具有较强的渗透能力,形成皱果苋耐旱性的内在基础.     

Osmotic imbalance in inositol-starved spheroplasts of Saccharomyces cerevisiae.

Physiological states associated with inositol starvation of spheroplasts of Saccharomyces cerevisiae were investigated and compared with conditions preceding death of starved whole cells. In the absence of synthesis of inositol-containing lipids, cell surface expansion terminated after one doubling of whole cells. In spheroplasts, cessation of membrane expansion was apparently followed by rapid development of an osmotic imbalance, causing lysis. Continued synthesis and accumulation of cytoplasmic constituents within the limited cell volume were implicated as a cause of the osmotic imbalance. In whole cells, an increase in internal osmotic pressure also follows termination of membrane and cell wall expansion. The cell wall prevents lysis, allowing a state of increasing cytoplasmic osmotic pressure to persist in the period preceding onset of inositol-less death.     

Osmotic adjustment in the filamentous fungus Aspergillus nidulans.

Aspergillus nidulans was shown to be xerotolerant, with optimal radial growth on basal medium amended with 0.5 M NaCl (osmotic potential [psi s] of medium, -3 MPa), 50% optimal growth on medium amended with 1.6 M NaCl (psi s of medium, -8.7 MPa), and little growth on medium amended with 3.4 M NaCl (psi s of medium, -21 MPa). The intracellular content of soluble carbohydrates and of selected cations was measured after growth on basal medium, on this medium osmotically amended with NaCl, KCl, glucose, or glycerol, and also after hyperosmotic and hypoosmotic transfer. The results implicate glycerol and erythritol as the major osmoregulatory solutes. They both accumulated during growth on osmotically amended media, as well as after hyperosmotic transfer, except on glycerol-amended media, in which erythritol did not accumulate. Furthermore, they both decreased in amount after hypoosmotic transfer. With the exception of glycerol, the extracellular osmotic solute did not accumulate intracellularly when mycelium was grown in osmotically amended media, but it accumulated after hyperosmotic transfer. It was concluded that the extracellular solute usually plays only a transient role in osmotic adaptation. The intracellular content of soluble carbohydrates and cations measured could reasonably account for the intracellular osmotic potential of mycelium growing on osmotically amended media.     

Osmotic mass transfer in the yeast Saccharomyces cerevisiae.

This paper reviews the passive mechanisms involved in the response of a yeast to changes in medium concentration and osmotic pressure. The results presented here were collected in our laboratory during the last decade and are experimentally based on the measurement of cell volume variations in response to changes in the medium composition. In the presence of isoosmotic concentration gradients of solutes between intracellular and extracellular media, mass transfers were found to be governed by the diffusion rate of the solutes through the cell membrane and were achieved within a few seconds. In the presence of osmotic gradients, mass transfers mainly consisting in a water flow were found to be rate limited by the mixing systems used to generate a change in the medium osmotic pressure. The use of ultra-rapid mixing systems allowed us to show that yeast cells respond to osmotic upshifts within a few milliseconds and to determine a very high hydraulic permeability for yeast membrane (Lp>6.10(-11) m x sec)-1) x Pa(-1)). This value suggested that yeast membrane may contain facilitators for water transfers between intra and extracellular media, i.e. aquaporins. Cell volume variation in response to osmotic gradients was only observed for osmotic gradients that exceeded the cell turgor pressure and the maximum cell volume decrease, observed during an hyperosmotic stress, corresponded to 60% of the initial yeast volume. These results showed that yeast membrane is highly permeable to water and that an important fraction of the intracellular content was rapidly transferred between intracellular and extracellular media in order to restore water balance after hyperosmotic stresses. Mechanisms implied in cell death resulting from these stresses are then discussed.     

Osmotic shrinkage of giant egg-lecithin vesicles.

Osmotic shrinkage of giant egg-lecithin vesicles was observed by phase-contrast microscopy. The vesicles remained or became spherical when shrinking. Small and thick-walled vesicles formed visible fingers attached to the sphere. The water permeability of the single bilayer was found to be 41 micrometers/s. A variety of observations indicate that osmosis induces a parallel lipid flow between the monolayers of the bilayer, leading to a strong positive spontaneous curvature. They also suggest the formation of mostly submicroscopic daughter vesicles. The estimated coupling constant, 2 . 10(-6) mol/mol, is large enough to be biologically significant.     

Osmotic shock of fertilized mouse ova.

The effect of osmotic changes on fertilized mouse ova was studied by measuring their survival, defined as development into hatching blastocysts, after exposure to various concentrations of ethanediol (ethylene glycol). In addition, a Boyle-van't Hoff plot was derived from exposing ova to hypotonic and hypertonic solutions ranging from 0.1 to 2.8 osmol. Volume of ova was inversely proportional to osmolality over this range. Extrapolation of this relationship yielded a nonosmotic volume of the ova of 22.5%. Eighty-five per cent or more of the ova survived exposure to this wide range of concentrations and developed into blastocysts. The rate of development of ova exposed to anisotonic solutions was the same as that of controls. Ova underwent osmotic shock when abruptly diluted out of concentrated solutions of ethanediol with an isotonic solution. Their survival was highly dependent on the ethanediol concentration with which they had equilibrated before dilution, and the manner, rate and temperature of dilution. The longer the exposure to ethanediol the greater was the sensitivity of the ova to osmotic shock, reflecting permeation of ethanediol into the ova. Osmotic shock could be alleviated by dilution at a high temperature, and prevented by the use of sucrose as an osmotic buffer at 37 degrees C. Identification of the variables that influence osmotic shock of ova will be helpful in the systematic study of their cryopreservation.     

Osmotic Adjustment and Osmotic Constituents in Roots of Mung Bean Seedlings

Osmotic adjustment in roots of mung bean seedlings (Vigna mungo(L.) Hepper) and the effect of cotyledon excision on the osmoticadjustment were investigated. The major osmotic constituentsin roots of intact seedlings were K⁺, Cl^–, free aminoacids and sugars (glucose, fructose and sucrose). All theseintracellular concentrations distinctly increased under osmoticstress and contributed to about 80% of the intracellular osmoticpressure of the root cell sap. Cotyledon excision remarkablysuppressed both the osmotic adjustment and the elongation inroots. However, the effect of cotyledon excision on intracellularK⁺ and Cl^– concentrations in roots was quite small. Twodifferent mechanisms are likely for the osmotic adjustment inroots. One is the K⁺ and Cl^–-dependent osmotic adjustmentwhich is cotyledon-independent, and the other is the osmoticadjustment dependent on the supply of free amino acids and sugarsfrom cotyledons. (Received September 20, 1986; Accepted January 14, 1987)     

Osmotic repression of anaerobic metabolic systems in Escherichia coli.

The influence of the osmolarity of the growth medium on anaerobic fermentation and nitrate respiratory pathways was analyzed. The levels of several enzymes, including formate dehydrogenase, hydrogenase, and nitrate reductase, plus a nickel uptake system were examined, as was the expression of the corresponding structural and regulatory genes. While some functions appear to be only moderately affected by an increase in osmolarity, others were found to vary considerably. An increase in the osmolarity of the medium inhibits both fermentation and anaerobic respiratory pathways, though in a more dramatic fashion for the former. fnr expression is affected by osmolarity, but the repression of anaerobic gene expression was shown to be independent of FNR regulatory protein, at least for hyd-17 and fdhF. This repression could be mediated by the intracellular concentration of potassium and is reversed by glycine betaine.     

Osmotic regulation of intracellular solute pools in Lactobacillus plantarum.

Bacteria respond to changes in medium osmolarity by varying the concentrations of specific solutes in order to maintain constant turgor pressure. The cytoplasmic pools of K+, proline, glutamate, alanine, and glycine of Lactobacillus plantarum ATCC 14917 increased when the osmolarity of the growth media was raised from 0.20 to 1.51 osmol/kg by KCL. When glycine-betaine was present in a high-osmolarity chemically defined medium, it was accumulated to a high cytoplasmic concentration, while the concentrations of most other osmotically important solutes decreased. These observations, together with the effects of glycine-betaine on the specific growth rate under high-osmolarity conditions, suggest that glycine-betaine is preferentially accumulated in L. plantarum. Uptake of glycine-betaine, proline, glutamate, and alanine was studied in cells that were alternately exposed to hyper- and hypo-osmotic stresses. The rate of uptake of proline and glycine-betaine increased instantaneously upon increasing the osmolarity, whereas that of other amino acids did not. This activation occurred also under conditions in which protein synthesis was inhibited was most pronounced when cells were pregrown at high osmolarity. The duration of net transport was a function of the osmotic strength of the assay medium. Glutamate uptake was not activated by an osmotic upshock, and the uptake of alanine was low under all conditions tested. When cells were subjected to osmotic downshock, a rapid efflux of accumulated glycine-betaine, proline, and alanine occurred whereas the pools of other amin acids remained unaffected. The results indicate that osmolyte efflux is, at least to some extent, mediated via specific osmotically regulated efflux systems and not via nonspecific mechanisms as has been suggested previously.     

Osmotic regulation of PhoE porin synthesis in Escherichia coli.

In Escherichia coli, adaptation to hyperosmotic conditions alters the expression of the outer membrane porins OmpF and OmpC. The amount of PhoE porin, which is normally induced by phosphate deprivation, was greatly reduced in cells adapted to high-osmolarity conditions. Osmoregulation of PhoE operated independently of the activity of the PhoR phosphate sensor and did not involve cross-talk from the homologous osmosensor EnvZ. PhoE synthesis was partially restored by additional copies of the positive regulator phoB+ and by the osmoprotectant glycine betaine.     

Osmotic significance of glycerol accumulation in exponentially growing yeasts.

Natural-abundance 13C-nuclear magnetic resonance spectroscopy has shown glycerol to be the major osmotically significant low-molecular-weight solute in exponentially growing, salt-stressed cells of the yeasts Saccharomyces cerevisiae, Zygosaccharomyces rouxii, and Debaromyces hansenii. Measurement of the intracellular nonosmotic volume (i.e., the fraction of the cell that is osmotically unresponsive) by using the Boyle-van't Hoff relationship (for nonturgid cells, the osmotic volume is directly proportional to the reciprocal of the external osmotic pressure) showed that the nonosmotic volume represented up to 53% of the total cell volume; the highest values were recorded in media with maximum added NaCl. Determinations of intracellular glycerol levels with respect to cell osmotic volumes showed that increases in intracellular glycerol may counterbalance up to 95% of the external osmotic pressure due to added NaCl. The lack of other organic osmotica in 13C-nuclear magnetic resonance spectra indicates that inorganic ions may constitute the remaining component of intracellular osmotic pressure.