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  • 1
    ISSN: 1432-2013
    Keywords: Intracellular pH ; SNARF-1 ; Carotid body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We report the use of a new pH-sensitive dualemission fluoroprobe, carboxy-seminaphthorhodafluor-1 (carboxy-SNARF-1) for ratiometric recording of intracellular pH (pHi) in small isolated cells. The method is illustrated with pHi measurement in single type-1 cells (cell diameter ∼10 μm) isolated from the carotid body of the neonatal rat. Carboxy-SNARF-1 is loaded using bath application of the acetoxymethyl ester. When excited at 540 nm, the fluoroprobe gives strong, inversely related emission signals at 590 nm and 640 nm. Stable ratiometric recordings of pHi can be achieved from a single cell (pHi 8.5-6.5) for up to 50 min. Photobleaching of the probe is minimised by illuminating at relatively low light intensity (50 W xenon lamp with 0.2% transmission neutral density filter). The probe can be calibrated in situ using the nigericin technique and this is in good quantitative agreement with the independent null-point technique (extracellular weak acid/weak base application) of Eisner et al. (1989). This fluoroprobe offers certain advantages over the other commonly used probe for pHi 2′,7′-bis-(2-carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF): (i) because of its two strong pH-sensitive peak emissions, SNARF displays a good signal-to-noise ratio for ratiometric recording at low light intensities; (ii) unlike BCECF, the dual emisson of SNARF requires no sequential mechanical switching of excitation filters, thus simplifying the epifluorescence set-up; (iii) because carboxy-SNARF-1 emission signals are at the yellow/red end of the visible spectrum, fluorescent drugs like amiloride, ethyl-isopropyl-amibride (EIPA), 4,4′-diisothiocyanostilbene 2,2′-disulphonic acid (DIDS) and cinnamate analogues do not interfere with the pHi recording, even when used at high concentrations.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 434 (1997), S. 429-437 
    ISSN: 1432-2013
    Keywords: Key words pHi ; SNARF ; Carotid body ; Type-1 cell ; Potassium-hydrogen exchange ; K+-H+ exchanger (KHE) ; Nigericin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Intracellular pH (pHi) was measured in enzymically isolated, neonatal rat carotid body type-1 cells, using the fluorophore carboxy-SNARF-1 (AM-loaded), and using the nigericin technique for in situ fluorescence calibration (nigericin is a membrane-soluble K+-H+ exchanger). In CO2/HCO3 –-free media, inhibiting Na+-H+ exchange produced a prompt fall of pHi (background acid-loading), the rate of which was reduced by raising the extracellular K+ concentration, [K+]o. pHi recovery from an intracellular acid or alkali load was also sensitive to changes of [K+]o. These results are similar to those of Wilding et al. (J Gen Physiol 100:593–608, 1992), who proposed the existence of an acid-loading, K+-H+ exchanger (KHE) in the type-1 cell. However, when nigericin was not used for post-experimental calibration, and the superfusion system was flushed exhaustively with strong detergent, alcohol and distilled water, then background acid-loading was attenuated, and the K+ o sensitivity of pHi insignificant. Background loading was increased again, and K+ o sensitivity restored, when cells were monitored in a superfusion system which had previously been exposed to a single nigericin-calibration protocol (followed by a short system wash with strong detergent and distilled water). We conclude that the previously reported expression of KHE in carotid body type-1 cells is an artefact caused by nigericin contamination. We have therefore quantified the pHi dependence of background loading in uncontaminated type-1 cells. We consider the possible implications of our work for reports of KHE in other cell types.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Carotid body ; Chemoreceptor ; Intracellular calcium ; Intracellular pH ; Extracellular pH ; Acidosis ; Hypercapnia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have investigated the effects of acidic stimuli upon [Ca2+]i in isolated carotid body type I cells from the neonatal rat using indo-1 (AM-loaded). Under normocapnic, non-hypoxic conditions (23 mM HCO3 −, 5% CO2 in air, pHo=7.4), the mean [Ca2+]i for single cells was 102±5.0 nM (SEM, n=55) with 58% of cells showing sporadic [Ca2+]i fluctuations. A hypercapnic acidosis (increase in CO2 to 10%–20% at constant HCO3 −, pHo 7.15–6.85), an isohydric hypercapnia (increase in CO2 to 10% at constant pHo=7.4) and an isocapnic acidosis (pHo=7.0, constant CO2) all increased [Ca2+]i in single cells and cell clusters. The averaged [Ca2+]i response to both hypercapnic acidosis and isohydric hypercapnia displayed a rapid rise followed by a secondary decline. The averaged [Ca2+]i response to isocapnic acidosis displayed a slower rise and little secondary decline. The rise of [Ca2+]i in response to all the above stimuli can be attributed to no single factor other than to a fall of pHi. The hypercapnia-induced rise of [Ca2+]i was almost completely abolished in Ca2+-free solution, suggesting a role for Ca2+ influx in triggering and/or sustaining the [Ca2+]i response. These results are consistent with a role for type I cell [Ca2+]i in mediating pH/PCO2 chemoreception.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Planta 157 (1983), S. 344-349 
    ISSN: 1432-2048
    Keywords: Glycinebetaine ; Ion, inorganic (distribution) ; Salt tolerance ; Shoot (solutes) ; Solute distribution ; Suaeda
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The distribution of sodium, potassium and glycinebetaine in shoot tissues of salt-treated Suaeda maritima was examined by semi-micro techniques after extraction into toluene-water. Much higher K/Na ratios were observed in the apical regions and in axillary buds than in more mature, fully vacuolated tissues. The younger tissues also contained very high levels of glycinebetaine. Electron-probe X-ray microanalysis of bulkfrozen and fractured preparations showed higher K/Na ratios and higher levels of sulphur and phosphorus in the cytoplasm of leaf primordial cells than in vacuoles of either young or old leaves, although the total counts were higher in the vacuolar samples. The results are discussed in relation to current models of subcellular solute compartmentation and salt tolerance in the Chenopodiaceae.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Planta 165 (1985), S. 392-396 
    ISSN: 1432-2048
    Keywords: Salt tolerance ; Osmotic adjustment ; Turgor ; Suaeda (salt tolerance)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Osmotic potentials and individual epidermal cell turgor pressures were measured in the leaves of seedlings of Suaeda maritima growing over a range of salinities. Leaf osmotic potentials were lower (more negative) the higher the salt concentration of the solution and were lowest in the youngest leaves and stem apices, producing a gradient of osmotic potential towards the apex of the plant. Epidermal cell turgor pressures were of the order of 0.25 to 0.3 MPa in the youngest leaves measured, decreasing to under 0.05 MPa for the oldest leaves. This pattern of turgor pressure was largely unaffected by external salinity. Calculation of leaf water potential indicated that the gradient between young leaves and the external medium was not altered by salinity, but with older leaves, however, this gradient diminished from being the same as that for young leaves in the absence of NaCl, to under 30% of this value at 400 mM NaCl. These results are discussed in relation to the growth response of S. maritima.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 82 (1991), S. 729 
    ISSN: 1432-2242
    Keywords: Triticum ; K/Na discrimination ; Salt tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A number of accessions of the three species of diploid wheat, Triticum boeoticum, T. monococcum, and T. urartu, were grown in 50 mol m-3 NaCl+2.5 mol m-3 CaCl2. Sodium accumulation in the leaves was low and potassium concentrations remained high. This was not the case in T. durum grown under the same conditions, and indicates the presence in diploid wheats of the enhanced K/Na discrimination character which has previously been found in Aegilops squarrosa and hexaploid wheat. None of the accessions of diploid wheat showed poor K/Na discrimination, which suggests that if the A genome of modern tetraploid wheats was derived from a diploid Triticum species, then the enhanced K/Na discrimination character became altered after the formation of the original allopolyploid. Another possibility is that a diploid wheat that did not have the enhanced K/Na discrimination character was involved in the hybridization event which produced tetraploid wheat, and that this diploid is now extinct or has not yet been discovered.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 87 (1993), S. 184-192 
    ISSN: 1432-2242
    Keywords: Tomato ; Salt tolerance ; Seed germination ; Isozyme markers ; QTL mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The germination responsiveness of an F2 population derived from the cross Lycopersicon esculentum (UCT5) x L. pennellii (LA716) was evaluated for salt tolerance at two stress levels, 150 mM NaCl + 15 mM CaCl2 and 200 mM NaCl + 20 mM CaCl2. Individuals were selected at both tails of the response distribution. The salt-tolerant and salt-sensitive individuals were genotyped at 16 isozyme loci located on 9 of the 12 tomato chromosomes. In addition, an unselected (control) F2 population was genotyped at the same marker loci, and gene frequencies were estimated in both selected and unselected populations. Trait-based marker analysis was effective in identifying genomic locations (quantitative trait loci, QTLs) affecting salt tolerance in the tomato. Three genomic locations marked by Est-3 on chromosome 1, Prx-7 on chromosome 3, and 6Pgdh-2 and Pgi-1 on chromosome 12 showed significant positive effects, while 2 locations associated with Got-2 on chromosome 7 and Aps-2 on chromosome 8 showed significant negative effects. The identification of genomic locations with both positive and negative effects on this trait suggests the likelihood of recovering transgressive segregants in progeny derived from these parental lines. Similar genomic locations were identified when selection was made either for salt tolerance or salt sensitivity and at both salt-stress treatments. Comparable results were obtained in uni- and bidirectional selection experiments. However, when marker allele gene frequencies in a control population are unknown, bidirectional selection may be more efficient than unidirectional selection in identifying marker-QTL associations. Results from this study are discussed in relationship to the use of molecular markers in developing salt-tolerant tomatoes.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 81 (1991), S. 321-326 
    ISSN: 1432-2242
    Keywords: Lycopersicon esculentum ; Salt tolerance ; Seed germination ; Maternal effects ; Tomato improvement ; Gene action
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The salt-tolerant cultivated tomato (Lycopersicon esculentum) accession, ‘PI174263’, and a sensitive cv, ‘UCT5’, were crossed to develop reciprocal F1, F2 and BC1 populations for genetic analysis of salt tolerance in tomatoes during seed germination. Variation was partitioned into embryo, endosperm and maternal (testa and cytoplasmic) components. Generation means analysis indicated that there were no significant embryo (additive, dominance or epistatic) effects on germination performance under salt stress. Highly significant endosperm additive and testa dominance effects were detected. The proportion of the total variance explained by the model containing these two components was R2=98.2%. Variance component analysis indicated a large genetic variance with additive gene action as the predominant component. Furhter inspection indicated that this variance was attributable to endosperm additive effects on germinability under salt stress. Narrow-sense heritability was estimated as moderately high. Implications for breeding procedures are discussed.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 89 (1985), S. 15-40 
    ISSN: 1573-5036
    Keywords: Elytrigia ; Epicuticular waxes ; Halophytes ; Leymus ; Potassium ; Roots ; Salt tolerance ; Shoots ; Sodium ; Transpiration ; Triticum ; Water use efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In the first part of this review the main features of salt tolerance in higher plants are discussed. The hypothesis of intracellular compartmentation of solutes is used as a basis for models of tolerance mechanisms operating in roots and in leaves. Consideration is given to the implications of the various mechanisms for the yield potential of salt-tolerant crop plants. Some work on the more salt-tolerant members of the Triticeae is then described. The perennial speciesElytrigia juncea andLeymus sabulosus survive prolonged exposure to 250 mol m−3 NaCl, whereas the annual Triticum species are severely affected at only 100 mol m−3 NaCl. In the perennial species the tissue ion levels are controlled within narrow limits. In contrast, the more susceptible wheats accumulate far more sodium and chloride than is needed for osmotic adjustment, and the effects of salt stress increase with time of exposure. Two different types of salt tolerance are exhibited in plants capable of growing at high salinities. In succulent Chenopodiaceae, for example, osmotic adjustment is achieved mainly by accumulation of high levels of sodium and chloride in the shoots, accompanied by synthesis of substantial amounts of the compatible solute glycinebetaine. This combination of mechanisms allows high growth rates, in terms of both fresh and dry weight. At the opposite end of the spectrum of salt tolerance responses are the halophytic grasses, which strictly limit the influx of salts into the shoots, but suffer from very much reduced growth rates under saline conditions. Another variation is shown in those species that possess salt glands. The development and exploitation of crop plants for use on saline soils is discussed in relation to the implications of these various mechanisms.
    Type of Medium: Electronic Resource
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