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  • Electronic Resource  (6)
  • 32.80.H  (2)
  • Addition lines  (2)
  • Benzamil  (2)
  • 1
    ISSN: 1432-2013
    Keywords: Key wordsXenopus oocytes ; Guinea-pig colon ; Na+ channel ; Single-channel analysis ; Amiloride ; Benzamil ; cAMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Guinea-pig distal colonic mRNA injection into Xenopus laevis oocytes resulted in expression of functional active epithelial Na+ channels in the oocyte plasma membrane. Poly(A)+ RNA was extracted from distal colonic mucosa of animals fed either a high-salt (HS) or a low-salt (LS) diet. The electrophysiological properties of the expressed amiloride-sensitive Na+ conductances were investigated by conventional two-electrode voltage-clamp and patch-clamp measurements. Injection of poly(A)+ RNA from HS-fed animals [from hereon referred to as HS-poly(A)+ RNA] into oocytes induced the expression of amiloride-sensitive Na+ conductances. On the other hand, oocytes injected with poly(A)+ RNA from LS-fed animals [LS-poly(A)+ RNA] expressed a markedly larger amount of amiloride-blockable Na+ conductances. LS-poly(A)+ RNA-induced conductances were completely inhibitable by amiloride with a K i of 77 nM, and were also blocked by benzamil with a K i of 1.8 nM. 5-(N-Ethyl-N-isopropyl)-amiloride (EIPA), even in high doses (25 μM), had no detectable effect on the Na+ conductances. Expressed amiloride-sensitive Na+ channels could be further activated by cAMP leading to nearly doubled clamp currents. When Na+ was replaced by K+, amiloride (1 μM) showed no effect on the clamp current. Single-channel analysis revealed slow gating behaviour, open probabilities (P o) between 0.4 and 0.9, and slope conductances of 3.8 pS for Na+ and 5.6 pS for Li+. The expressed channels showed to be highly selective for Na+ over K+ with a permeability ratio P Na/P K 〉 20. Amiloride (500 nM) reduced channel P o to values 〈 0.05. All these features make the guinea-pig distal colon of LS-fed animals an interesting mRNA source for the expression of highly amiloride-sensitive Na+ channels in Xenopus oocytes, which could provide new insights in the regulatory mechanism of these channels.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2242
    Keywords: Key words Aegilops markgrafii ; Triticum aestivum ; Addition lines ; Chromosome markers ; Homoeology ; Wheat ; Wheat microsatellites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We describe the use of wheat microsatellites for the discrimination of Aegilops markgrafii chromosomes. Twenty out of eighty eight wheat microsatellites (WMS) tested were able to distinguish Triticum aestivum-Ae. markgrafii addition lines. Six, three, three, one and six of 18 WMS can be used as markers for single Ae. markgrafii chromosomes B, C, D, F and G, respectively. Addition line A is not available but additional bands, appearing only in Ae. markgrafii and the T. aestivum-Ae. markgrafii amphiploid and not in any of the available addition lines, indicate that three WMS detect markers for Ae. markgrafii chromosomes A. Addition line E could not be detected by any of the WMS markers applied, although the 20 WMS represented all the homologous groups of wheat. All three WMS located on the short arm of group-2 chromosomes were located on Ae. markgrafii chromosome B; three of four WMS, located on the long arm of wheat group-2 chromosomes, were specific to Ae. markgrafii chromosome G and three of four WMS, specific to group-5 chromosomes, were markers for Ae. markgrafii chromosome C, indicating the homoeology of these wheat chromosome arms with the respective Ae. markgrafii chromosomes.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2242
    Keywords: Key words Aegilops markgrafii ; Triticum aestivum ; RAPD ; Addition lines ; Leaf rust ; Powdery mildew
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Aegilops markgrafii contains resistance genes to powdery mildew, leaf rust and stripe rust, and also has high crude protein and lysine contents, which can be useful for wheat improvement. These important traits are localized on different chromosomes. Disomic Triticum aestivum-Ae. markgrafii addition lines and euploid introgression lines showing leaf-rust and powdery mildew resistance were screened with RAPDs to detect chromosome-specific markers which can accelerate the breeding process. RAPD markers for all six available disomic addition lines were obtained. The additional chromosomes B, C, D, E, F and G were identified by three, three, three, two, one and seven primers, respectively. All three chromosome-B-specific RAPD markers demonstrated the presence of alien chromatin in the leaf-rust-resistant 42-chromosome introgression lines as well as in the segregating progeny. The three chromosome-C-identifying primers also demonstrated the presence of that chromosome in powdery mildew-resistant euploid introgression lines. The substitution lines (5A)5C and (5D)5C with different genetic backgrounds for both parents, in comparison to the lines mentioned above, showed the chromosome C-specific band with only two of the three primers. The chromosome F-specific primer and a primer evident on all the Ae. markgrafii chromosomes analysed did not generate the expected fragments on the chromosome Fdel addition line, indicating that the markers are located on the deleted part of chromosome F.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 163 (1993), S. 153-159 
    ISSN: 1432-136X
    Keywords: Na+ channels ; Amiloride ; Benzamil ; Noise analysis ; Leech, Hirudo medicinalis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The dorsal skin of the leech Hirudo medicinalis was used for electrophysiological measurements performed in Ussing chambers. The leech skin is a tight epithelium (transepithelial resistance = 10.5±0.5 kΩ· cm-2) with an initial short-circuit current of 29.0±2.9 μA·cm-2. Removal of Na+ from the apical bath medium reduced short-circuit current about 55%. Ouabain (50μmol·l-1) added to the basolateral solution, depressed the short-circuit current completely. The Na+ current saturated at a concentration of 90 mmol Na+·l-1 in the apical solution (K M=11.2±1.8 mmol·l-1). Amiloride (100 μmol·l-1) on the apical side inhibited ca. 40% of the Na+ current and indicated the presence of Na+ channels. The dependence of Na+ current on the amiloride concentration followed Michaclis-Menten kinetics (K i=2.9±0.4 μmol·l-1). The amiloride analogue benzamil had a higher affinity to the Na+ channel (K i=0.7±0.2 μmol·l-1). Thus, Na+ channels in leech integument are less sensitive to amiloride than channels known from vertebrate epithelia. With 20 mmol Na+·l-1 in the mucosal solution the tissue showed an optimum amiloride-inhibitable current, and the amiloride-sensitive current under this condition was 86.8±2.3% of total short-circuit current. Higher Na+ concentrations lead to a decrease in amiloride-blockade short-circuit current. Sitmulation of the tissue with cyclic adenosine monophosphate (100 μmol·l-1) and isobutylmethylxanthine (1 mmol·l-1) nearly doubled short-circuit current and increased amiloride-sensitive Na+ currents by 50%. By current fluctuation analysis we estimated single Na+ channel current (2.7±0.9 pA) and Na+ channel density (3.6±0.6 channels·μm-2) under control conditions. After cyclic adenosine monophosphate stimulation Na+ channel density increased to 5.4±1.1 channels·μm-2, whereas single Na+ channel current showed no significant change (1.9±0.2 pA). These data present a detailed investigation of an invertebrate epithelial Na+ channel, and show the similarities and differences to vertebrate Na+ channels. Whereas the channel properties are different from the classical vertebrate Na+ channel, the regulation by cyclic adenosine monophosphate seems similar. Stimulation of Na+ uptake by cyclic adenosine monophosphate is mediated by an increasing number of Na+ channels.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    The European physical journal 16 (1990), S. 97-106 
    ISSN: 1434-6079
    Keywords: 34.80.D ; 32.80.H
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract We have measured the ratio of cross sections σ(KL 2,3)/σ(K) for neon for electron impact in the energy range ofE 0=1.5 ... 50 keV via the intensity ofKL 2,3−LLL 2,3 Auger satellite lines relative to the intensity ofKL 1 L 2,3 (3 P) diagram line. The experimental ratio decreases over the full range of energyE 0 which is contrary to an earlier result by Carlson et al. We have also measured the alignment ofKL 2,3 1 P and3 P states via the angular distribution of Auger satellite intensity for the energy rangeE 0=1.5 ... 4 keV, within experimental error we have found a zero alignment. The totalK Auger spectrum, measured forE 0=40 keV and at the magic angle of emission ϑ=54.7°, has been decomposed into its components by using appropriate line shapes distorted by postcollision interaction. Finally, we discussed whether the lines observed at the high-energy side ofKL 2,3−LLL 2,3 Auger satellite lines can be interpreted as structures caused by an angular momentum exchange in the postcollision interaction predicted by Niehaus and Zwakhals.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    The European physical journal 31 (1994), S. 53-54 
    ISSN: 1434-6079
    Keywords: 34.80.D ; 32.80.H
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract From intensity of Auger transitions (1s2p)−1 1 P,3 P→2p −3 2 P,2 D+e A − relative to 1s −1 2 S→2p −2 1 D+e A − measured at ϑ=54 and 90° relative to the primary electron beam we have determined the alignment of double vacancy states (1s 2p)−1 1 P and3 P of neon for electron impact ionization for the impact energies 1.5, 2.0, 2.5 and 4.0 keV. ForE 0=1.5, 2.0 and 2.5 keV the alignment is compatible with zero with an upper limit of , forE 0=4.0 keV a small negative value was found.
    Type of Medium: Electronic Resource
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