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  • 1
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Biomembranes 978 (1989), S. 134-144 
    ISSN: 0005-2736
    Keywords: (Rabbit) ; Apical membrane ; Patch clamp ; Potassium ion channel ; Proximal tubule
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Biomembranes 938 (1988), S. 257-269 
    ISSN: 0005-2736
    Keywords: (Rabbit kidney) ; Monocarboxylate carrier ; Proximal tubule ; Sodium ion cotransport ; intracellular ; pH
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Biomembranes 1070 (1991), S. 387-400 
    ISSN: 0005-2736
    Keywords: (Rabbit) ; Patch clamp ; Potassium ion conductance
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Biochimica et Biophysica Acta (BBA)/Biomembranes 817 (1985), S. 333-342 
    ISSN: 0005-2736
    Keywords: (Frog urinary bladder) ; Colchicine ; Cytochalasin B ; Freeze-fracture ; Oxytocin ; Water permeability
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Medicine , Physics
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Techniques using microdissected tubules from rabbit kidney allow the isolation of well defined segments which can be cultured, to obtain pure renal cell epithelia. From microdissected proximal tubules, we obtained epithelia the cells of which exhibit some of the antigenic expressions of the initial proximal cells. For this purpose, we used three monoclonal antibodies raised against apical brush border membranes of the proximal tubules. We determined with precision the identity and some of the molecular characteristics of the antigens bound by these three antibodies and found that they correspond to three hydrolases present in the brush borders of proximal renal cells (amino-peptidase, dipeptidyl-peptidase IV and endopeptidase). These apical markers are expressed by the growing cells of primary cultures from proximal tubules, suggesting strongly that they are effectively proximal cells and that no appreciable dedifferentiation occured during the growth process. We have also shown that apical expression of these hydrolases on the plasma membrane of the epithelium occured only after several days of culture and determined the complete polarization of the cells. Electron microscopy studies confirmed the degree of polarization of the cultured cells by the presence of numerous microvilli on their apical face.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2013
    Keywords: Proximal tubule ; Primary culture ; Immunological characterization ; Patch clamp ; Ionic channel ; Ca2+ sensitivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Primary cultures were obtained from microdissected rabbit proximal tubules (S1 segments). The growing epithelia were maintained in culture for up to 30 days. Electron microscopy study revealed that the cells formed a monolayer and showed a morphological polarity with apical microvilli and tight junctions. An immunofluorescence technique using two monoclonal antibodies raised against two apical brush border enzymes of the proximal tubule (LAP, DPP IV) revealed that these enzymes were expressed in the cultured cells. Membrane associated and cytosolic enzyme activities were measured on 12, 20 and 30-day-old cultures. Cultured epithelia exhibited leucine aminopeptidase, γ glutamyl transferase and fructose 1–6 biphosphatase activities that remained constant for up to 30 days, whereas alkaline phosphatase activity decreased in the oldest cultures. Hexokinase activity on the other hand, increased after 12 days of culture. Cyclic AMP synthesis was stimulated by parathyroid hormone at 12, 20 and 30 days of culture and was insensitive to arginine vasopressin. After 20 days of culture the epithelia grown on permeable supports developed a transepithelial potential of −0.13 mV (apical negative) and a transepithelial resistance of 37 Ω cm2 that increased to −1.13 mV and 60 Ω cm2 respectively in 30-day-old cultures. The patch clamp technique was applied to the apical membrane of 12–15-day-old cultures. In the whole cell recording configuration, a cellular potential of −61.5 mV was measured, which was mainly due to K+ diffusion. A non-selective cationic channel was present in the apical membrane of the cultured cells. In cell-attached patches the channel carried an inward current and had a conductance of 13 pS. On excised patches the channel discriminated poorly between Na+ and K+ and was impermeant to Cl− and its conductance ranged between 20 and 28 pS. The channel activity was not voltage dependent but required a high calcium concentration (1 mM Ca2+) on the cytoplasmic face.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2013
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Calcium is actively reabsorbed in the distal nephron segments and recent studies have demonstrated the presence of Ca2+ channels in these epithelial cells, which could be involved in transepithelial transport. To test this possibility, single-channel currents were recorded by the patch-clamp technique in the apical membrane of primary cultures of the rabbit distal bright convoluted tubule cells (DCTb). In the cell-attached mode with 100 mmol/l BaCl2 in the pipette and 145 mmol/l NaCl in the bath, inward negative currents, consistent with Ba2+ currents, were recorded. In these conditions, the single-channel conductance was 15 pS. In excised insideout patches, the single-channel conductance was 13 pS and the current reversal potential of +60 mV was close to the Nernst equilibrium potential for Ba2+ (〉+58 mV). Similar experiments conducted with Ca2+ as the main charge carrier showed that this ion was less permeant through the channel than Ba2+ (P Ba/P Ca≈1.4). We also showed that the Ca2+-channel blocker, lanthanum (1 μmol/l La3+), added on the cytosolic side of the membrane, reversibly blocked the channel activity. On the other hand, verapamil (0.1 mmol/l) and nifedipine (10 μmol/l), perfused on the cytosolic side of the membrane, abolished the channel activity but this effect was not reversible. Another type of channel was also identified in the apical membrane of cultured DCTb cells. Ion-substitution experiments showed that this 21-pS conductance channel did not discriminate between Na+ and K+ and did not conduct Ba2+. 4′-Methyl-2-diphenylamine-carboxylic acid (10 μmol/l), added on the cytosolic side of the membrane, reversibly blocked this cationic channel whereas La3+ (10(μmol/l) had no effect on its activity. We conclude that (a) La3+-sensitive Ca2+ channels are present in the apical membrane of DCTb cells and may represent the apical uptake pathway in the transepithelial calcium transport. (b) These Ca2+ channels are distinct from the non-selective cationic channels that do not show Ba2+ permeability.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-1424
    Keywords: Key words: Intercalated cells — Cortical collecting tubule — Acidification — Cl−/HCO−3 exchange — Plasticity of transport — Polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. High speed video imaging microscopy and the pH-sensitive fluorophore2′,7′,-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF) were used to examine acid-base functions of beta-intercalated cells of the rabbit cortical collecting duct. The presence of intercalated cells was established and the properties of apical and basolateral acid-base transporters assessed by monitoring cell pH during acid loading and luminal and basolateral ion substitutions. We showed that treatment of beta-intercalated cells with ammonium chloride (20 mm) induced a profound decrease of their intracellular pH from 6.98 ± 5.93 ± 0.08. pH recovery occurred after different lag periods ranging between 2 to 15 min (0.22 ± 0.04 dpH/dt). We demonstrated that this pH recovery mechanism was independent of basolateral Na+ and apical HCO− 3 and K+. It was also not affected by apical and basolateral addition of NEM, by basolateral DIDS and by apical application of the H-KATPase inhibitor SCH28080. The process of pH recovery was however, critically dependent on basolateral HCO− 3. These results are best explained by acid-induced insertion and/or activation of chloride-bicarbonate exchangers that are functional properties with their apical analogues.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 176 (2000), S. 151-158 
    ISSN: 1432-1424
    Keywords: Key words: Proestrus — Estrus — Osmotic Permeability — Aquapoins — HgCl2, Diabetes mellitus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. Important functional and structural modifications occur in mammalian oocytes during their arrival to maturity. In this process, oocytes switch from a high activity level, implying an important metabolic rate and a coordinated movement of water and solutes, to a lower functional state. The aim of this work was to study the mechanisms involved in water movements during oocyte arrival to maturity. Volume changes, induced by an osmotic gradient, were followed by video microscopy in rat oocytes. The water osmotic permeability (P osm ) of immature oocytes (proestrus) was sensitive to HgCl2 and phloretin. In contrast, mature oocytes (estrus) had a reduced P osm that was not sensitive to these compounds. When proestrus oocytes were incubated in vitro at 37°C they spontaneously arrived at maturity and its P osm decreased between four and six hours of incubation. RT-PCR experiments were performed using specific primers for all rat aquaporins that had been cloned. We found that aquaporin-9 transcript (AQP9) is present in proestrus oocytes but not in estrus oocytes. AQP9 has been recently described as a ``broad selective channel'' responsible for solute and water transfers in highly active cells. Our experiments showed that proestrus oocytes, but not estrus, are permeable to mannitol. It is concluded that during the process of maturation, P osm decreases and AQP9 transcripts disappear. We report here the first study correlating water permeability and aquaporin mRNA expression in mammalian oocytes.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 86 (1985), S. 239-245 
    ISSN: 1432-1424
    Keywords: water channels ; glutaraldehyde fixation ; frog urinary bladder ; unstirred layers ; osmotic and diffusional permeabilities ; Rana esculenta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Unidirectional and net water movements were determined, in frog urinary bladders, before and after glutraldehyde fixation. Experiments were performed in three experimental conditions: 1) in nonstimulated preparations, 2) after the action of antidiuretic hormone (ADH) and 3) in nonstimulated preparations to which amphotericin B was incorporated from the luminal bath. As previously observed for net water fluxes, the increase in the unidirectional water movement induced by ADH was well preserved by glutaraldehyde fixation. After correction for the effects of unstirred layers and nonosmotic pathways, the observed correlation between the ADH-induced increases in the osmotic (Pf) and diffusional (Pd) permeability coefficients was not modified by the fixative action (before glutaraldehyde: slope 11.19,r:0.87±0.07;n=12; after glutaraldehyde: slope 10.67,r:0.86±0.04,n=39). In the case of amphotericin B, ΔPf/ΔPd=3.08 (r: 0.83±0.08), a value similar to that observed in lipid bilayers or in nonfixed toad urinary bladders. It is concluded that 1) The experimental approach previously employed to study water channels in artificial lipid membranes and in amphibian urinary bladders, can be applied to the glutaraldehyde-fixed frog urinary bladder. 2) Glutaraldehyde fixation does not modify the permeability properties of the ADH-induced water channels. 3) Any contribution of exo-endocytic processes or cell regulatory mechanisms to the observed permeability parameters can probably be excluded. 4) Glutaraldehyde-fixed preparations are a good model to characterize these water pathways.
    Type of Medium: Electronic Resource
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