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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 60 (1982), S. 1103-1113 
    ISSN: 1432-1440
    Keywords: Proximal tubule ; Thick ascending limb ; Henle's loop ; Membrane transport ; Transport alteration ; Diuretics ; Proximaler Tubulus ; dicker aufsteigender Teil ; Henle'sche Schleife ; Membrantransport ; Transportstörung ; Diuretika
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Die Kenntnisse der renalen Transportmechanismen sind während den letzten Jahren dank den Fortschritten in der Methodik enorm angewachsen. Der transepitheliale Transport sowie seine pathologische und pharmakologische Veränderung können teilweise als Membranphänomene verstanden werden. Am Beispiel natriumgekoppelter Transportmechanismen im proximalen Tubulus sowie des Natrium-chlorid-Transportes in der dicken aufsteigenden Henle'schen Schleife werden in diesem Artikel Membrantransportmechanismen diskutiert. Es wird gezeigt, daß geänderte Nierenfunktion oft mit geändertem Membrantransport gleichzusetzen ist.
    Notes: Summary Over the past few years, our knowledge on renal tubular transport mechanisms has increased considerably. Due to new technical developments, it is now possible to understand in part transepithelial transport and its pathological and pharmacological alterations at the level of the cell membranes. Different membrane transport mechanisms are discussed in this article, whereby sodium coupled solute transport in the proximal tubule and sodium chloride transport in the thick ascending limb of Henle's loop are taken as examples. It is indicated that an altered function of the kidney can often be equated with an alteration of the membrane transport.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 57 (1979), S. 977-991 
    ISSN: 1432-1440
    Keywords: Epithelial transport ; Kidney ; Small intestine ; Electrolyte ; Epithelialer Transport ; Niere-Darm-Elektrolyt ; Elektrochemisches Gradient
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Das Epithel von Niere und Darm besteht aus einer einzigen Lage von Zellen, die an ihrer luminalen Seite durch Schlußleisten zusammengekittet sind. Der Stofftransport geht entweder transzellulär durch die Zellen hindurch und ist dann in der Regel aktiv, oder er geht parazellulär an den Zellen vorbei durch die Schlußleisten und interzellulären Spalten und ist dann passiv. Die Triebkraft für den aktiven Transport kommt entweder direkt aus dem Stoffwechsel und wirkt mittels ATPasen auf die zu transportierenden Stoffe. Wir haben dann einen primär aktiven Transport vor uns. Oder sie kommt aus Gradienten von Substanzen, in erster Linie Natriumionen, die ihrerseits primär aktiv transportiert wurden. Man spricht dann von sekundär aktivem Transport. Die Triebkräfte für den passiven Transport sind Konzentrations- bzw. elektrochemische Potentialdifferenzen sowie der durch Reibung bedingte Mitreißeffekt des resorbierten Wassers. Sowohl in Niere als auch im Darm haben die proximalen Abschnitte, wo eine große Flüssigkeitsmenge isoton resorbiert wird, undichte Schlußleisten, so daß eine beträchtliche Substanzmenge passiv resorbiert werden kann. In den distalen Abschnitten hingegen, wo der Transport geregelt wird, sind die Schlußleisten dicht, so daß entsprechende Konzentrationsunterschiede erzeugt und aufrecht erhalten werden können. Aktiver Transport durch die Epithelzellen hindurch ist indessen nur möglich, wenn der Stofftransport polar ist, d.h. an der luminalen Zellseite anders als an der kontraluminalen Zellseite. Durch elektrophysiologische Messungen an den einzelnen Zellseiten als auch durch Transportmessungen an geschlossenen Vesikeln, die von den beiden Zellseiten gewonnen wurden, konnten die treibenden Kräfte für die einzelnen Substanzen weitgehend festgelegt werden. An Schemata, in die die Transportmechanismen der einzelnen Zellseiten eingezeichnet sind, wird eine weitgehende Identität der Transportmechanismen im proximalen Tubulus und Dünndarm deutlich.
    Notes: Summary Epithelia of kidney and small intestine consist of one layer of cells which, at their luminal edge, are linked together by terminal bars. Solute transport proceeds either across the cells, which is true of all active transports, or it proceeds paracellularly through the basolateral spaces and terminal bars and is then passive. The driving force for the active transport of a substance is derived either directly from metabolism (primary active transport), or from the gradient of another solute, usually Na+, which in turn is created by primary active transport. In the latter case the transport is referred to as secondary active. The driving forces of passive transport are the electrochemical gradient of the respective substance and solvent drag. The proximal parts of the kidney as well as of the intestine are leaky so that a considerable part of net reabsorption proceeds passively. Their distal parts, however, where the transport is regulated, are tight so that large concentration differences can be created and maintained. Transcellular active transport is only possible if the cells are polar, i.e., the transport characteristics of the luminal cell membrane differ from those of the contraluminal cell membrane. By measuring the cellular electrical potential difference or by measuring transport into isolated plasma membrane vesicles from either cell side the driving forces for the two transport steps, the luminal and contraluminal, have been elucidated. Schemes for the transport steps in the proximal tubule and in the small intestine are given. They show the principal similarity of the transport of many substances in both epithelia.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 64 (1986), S. 824-828 
    ISSN: 1432-1440
    Keywords: Proximal tubule ; Sodium phosphate cotransport ; cAMP ; Protein kinases ; Renal tissue culture ; Proximaler Tubulus ; Natrium-Phosphat-Kotransport ; cAMP ; Proteinkinasen ; Nieren-Gewebekultur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Die Kenntnisse der intrazellulären Regulationsmechanismen in Hormon-kontrollierten Zell-Funktionen sind während der letzten Jahre enorm angewachsen. Die Besetzung des Rezeptors in der Plasma-Membran führt zur Bildung verschiedener intrazellulärer Signale („Messenger“), z.B. cyclische Nucleotide (cAMP, cGMP), Inositoltrisphosphate (IP3), Diacylglycerin (DAG), Erhöhung der Kalzium-Konzentration. Diese „Messenger“ kontrollieren die Aktivität verschiedener Regulationsmechanismen, die entweder sich folgend oder parallel schlußendlich zur biologischen Antwort führen. Bei der PTH-abhängigen Regulation des Phosphattransportes sind cAMP-abhängige und Kalzium-abhängige Regulationsmechanismen beteiligt: Versuche an kultivierten Epithelzellen haben bestätigt, daß die Stimulierung der Adenylatcyclase als erstes Ereignis auftritt. Das cAMP Signal kann jedoch umgangen werden und eine direkte Aktivierung der Proteinkinase C kann die PTH-Hemmung nachahmen. Als letztes Ereignis in der Regulationskette tritt vermutlich eine Entfernung des Transportsystemes auf.
    Notes: Summary The knowledge about intracellular regulatory cascades in hormone action has increased considerably over the last few years. Receptor occupation at the plasma membrane level results in a production of intracellular messengers, such as cyclic nucleotides (cAMP, cGMP), inositoltrisphosphate (IP3), diacylglycerol (DAG) and a rise in cytosolic calcium concentration. These messengers control the activity of different regulatory mechanisms which operate either in sequence or in parallel to generate the final biological response. In PTH-dependent regulation of renal phosphate transport, cAMP-dependent and calcium-dependent mechanisms are involved: Recent experiments with cultured renal epithelial cells have confirmed that activation of adenylate cyclase is the initial event. However, the cAMP signal can be bypassed and direct activation of protein kinase C seems to mimic PTH induced inhibition of phosphate transport. The final event in the regulatory cascade is most likely a removal of the phosphate transport system followed by a degradation.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 151 (1996), S. 215-224 
    ISSN: 1432-1424
    Keywords: Key words: Amino acid transport — Brush-border membrane — Intestine — Epithelium — Caco-2 cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. The nature of transepithelial and cellular transport of the dibasic amino acid lysine in human intestinal epithelial Caco-2 cells has been characterized. Intracellular accumulation of lysine across both the apical and basolateral membranes consists of a Na+-independent, membrane potential-sensitive uptake. Na+-independent lysine uptake at the basolateral membrane exceeds that at the apical membrane. Lysine uptake consists of both saturable and nonsaturable components. Na+-independent lysine uptake at both membranes is inhibited by lysine, arginine, alanine, histidine, methionine, leucine, cystine, cysteine and homoserine. In contrast, proline and taurine are without inhibitory effects at both membranes. Fractional Na+-independent lysine efflux from preloaded epithelial layers is greater at the basolateral membrane and shows trans-stimulation across both epithelial borders by lysine, arginine, alanine, histidine, methionine, and leucine but not proline and taurine. Na+-independent lysine influx (10 μm) in the presence of 10 mm homoserine shows further concentration dependent inhibition by lysine. Taken together, these data are consistent with lysine transport being mediated by systems bo,+, y+ and a component of very low affinity (nonsaturable) at both membranes. The relative contribution to lysine uptake at each membrane surface (at 10 μm lysine), normalized to total apical uptake (100%), is apical bo,+ (47%), y+ (27%) and the nonsaturable component (26%), and basal bo,+ (446%), y+ (276%) and the nonsaturable component (20%). Northern analysis shows hybridization of Caco-2 poly(A)+RNA with a human rBAT cDNA probe.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1424
    Keywords: Key words: Adenosine receptors — A6 cells — Chloride channels — Sodium transport — cAMP — Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. The effect of adenosine regulation on sodium and chloride transport was examined in cultured A6 renal epithelial cells. Adenosine and its analogue N6-cyclopentyladenosine (CPA) had different effects on short-circuit current (I sc) depending on the side of addition. Basolateral CPA addition induced an approximately threefold increase of the I sc that reached a maximum effect 20 min after addition and was completely inhibited by preincubation with either an A2 selective antagonist, CSC, or the sodium channel blocker, amiloride. Apical CPA addition induced a biphasic I sc response characterized by a rapid fourfold transient increase over its baseline followed by a decline and a plateau phase that were amiloride insensitive. The A1 adenosine antagonist, CPX, completely prevented this response. This I sc response to apical CPA was also strongly reduced in Cl−-free media and was significantly inhibited either by basolateral bumetanide or apical DPC preincubation. Only basolateral CPA addition was able to induce an increase in cAMP level. CPA, added to cells in suspension, caused a rapid rise in [Ca2+] i that was antagonized by CPX, not affected by CSC and prevented by thapsigargin preincubation. These data suggest that basolateral CPA regulates active sodium transport via A2 adenosine receptors stimulating adenylate cyclase while apical CPA regulates Cl− secretion via A1 receptor-mediated changes in [Ca2+] i .
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-1424
    Keywords: Key words: Renal Na/Pi-cotransport — Renal Na/sulfate-cotransport — Sf9 cells — Purification — Ni2+-chelate chromatography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. Two mammalian sodium-dependent anion-cotransporters (NaPi-2 for phosphate and NaSi-1 for sulfate) have been expressed in Sf9 insect cells using the baculovirus expression system. A histidine tag was introduced at the C-termini in order to facilitate purification by metal-affinity chromatography. Sf9 cells infected with the histidine-tagged Ni/P i -cotransporter exhibited more than 60-fold higher sodium-dependent transport of phosphate compared to noninfected cells. Expressed Na/P i -cotransport exhibited a K m of P i of 0.21 mm and an apparent K m of sodium of 92 mm. Infected cells expressed a 65 kDa polypeptide as detected by Western blotting and immunoprecipitation. Sf9 cells infected with the histidine-tagged NaSi-1 or untagged NaSi-1 protein expressed sodium-dependent sulfate cotransport up to 60-fold higher compared to noninfected cells. Transport of sulfate was highly dependent on sodium exhibiting a K m of SO2− 4 of about 0.3–0.4 mm and a K m of sodium of 55 mm. By Western blotting and immunoprecipitation expressed NaS i -1 proteins were detected at 55–60 kDa. These studies demonstrate that histidine tagged proximal tubular Na-dependent cotransporters for phosphate and sulfate can be expressed functionally in Sf9 cells and that the kinetic characteristics were not altered by the introduction of a histidine tag at the C-termini. Furthermore, it is demonstrated that after solubilization under denaturing conditions histidine-tagged cotransporter proteins can be purified by metal-chelate affinity chromatography.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-1424
    Keywords: Key words: Na/Pi-cotransporter — PTH — Endocytosis — Tyrosine-based signals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. The rat renal type II Na/Pi-cotransporter (NaPi2), which is regulated by mechanisms involving endocytosis and lysosomal degradation, contains two sequences that show high homology with two tyrosine (Y)-based consensus motifs previously reported to be involved in such intracellular trafficking: GY402FAM matching the consensus sequence GYXXZ, and Y509RWF matching the motif YXXO. Mutations of any of these two Y nearly abolished the NaPi2 mediated 32P i -uptake after cRNA-injection into oocytes. The mechanisms underlying these defects are however different. Mutation of the Y402 results in a lack of glycosylation and reduced surface expression of the cotransporter, that are specific for the Y402 mutation since substitution of the neighboring F403 did not have any effect. The inhibitory effect of the Y509 mutation is related to a functional inactivation of the protein expressed in the plasma membrane; mutation of the neighboring R510 also led to a decrease in the cotransporter activity. Pharmacological activation of the protein kinase C cascade by DOG induced the retrieval of both wild-type (WT) as well as Y509 cotransporters from the oocyte plasma membrane. These data suggest that the Y402 is important for the surface expression whereas Y509 for the function of the type II Na/P i -cotransporter expressed in oocytes. Y509 seems not to be involved in the membrane retrieval of the cotransporter.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 144 (1995), S. 43-48 
    ISSN: 1432-1424
    Keywords: Renal Na/P i -cotransport ; Sf9 cells ; Infection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The recently cloned Na/P i -cotransport system NaPi-2 is an apical membrane protein of rat proximal tubular cells and is involved in proximal phosphate reabsorption. To make the protein available for further functional/structural studies, this transport system has been expressed in Sf9 insect cells using a recombinant baculovirus. Sf9 cells infected with NaPi-2 (or 6His tagged NaPi-2) expressed functional Na/P i -cotransport up to 20- to 50-fold over noninfected Sf9 cells. Transport of phosphate in infected cells was highly dependent on sodium, exhibited a K m for P i of 0.114 mm and an apparent K m for Na of 63 mm (Hill coefficient of approximately 3) and was stimulated by high external pH. Infected cells expressed a polypeptide of 65 kDa representing a nonglycosylated form of the 85 kDa mature NaPi-2 transporter as present in proximal tubular brush-border membranes. By confocal microscopy expression of NaPi-2 protein was observed in the plasma membrane, yet submembranous accumulation of NaPi-2 protein could not be excluded. This demonstrates that the rat proximal tubular Na/P i -cotransport system NaPi-2 can be successfully expressed in Sf9 cells with characteristics similar to that in isolated brush-border membranes. The 6His tagging will permit isolation of the NaPi-2 cotransporter in amounts sufficient for structural/functional studies.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 154 (1996), S. 1-9 
    ISSN: 1432-1424
    Keywords: Key words: Phosphate transport — Dietary phosphate — Parathyroid hormone — Endocytosis — Exocytosis — Microtubules--〉
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-1424
    Keywords: Key words: Cl− Channel — Organic Anions — Phosphate — NPPB
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract. Expression of the protein NaPi-1 in Xenopus oocytes has previously been shown to induce an outwardly rectifying Cl− conductance (GCl), organic anion transport and Na+-dependent P i -uptake. In the present study we investigated the relation between the NaPi-1 induced GCl and P i -induced currents and transport. NaPi-1 expression induced P i -transport, which was not different at 1–20 ng/oocyte NaPi-1 cRNA injection and was already maximal at 1–2 days after cRNA injection. In contrast, GCl was augmented at increased amounts of cRNA injection (1–20 ng/oocyte) and over a five day expression period. Subsequently all experiments were performed on oocytes injected with 20 ng/oocytes cRNA. P i -induced currents (Ip) could be observed in NaPi-1 expressing oocytes at high concentrations of P i (≥ 1 mm P i ). The amplitudes of Ip correlated well with GCl. Ip was blocked by the Cl− channel blocker NPPB, partially Na+-dependent and completely abolished in Cl− free solution. In contrast, P i -transport in NaPi-1 expressing oocytes was not NPPB sensitive, stronger depending on extracellular Na+ and weakly affected by Cl− substitution. Endogenous P i -uptake in water-injected oocytes amounted in all experiments to 30–50% of the Na+-dependent P i -transport observed in NaPi-1 expressing oocytes. The properties of the endogenous P i -uptake system (K m for P i 〉 1 mm; partial Na+- and Cl−-dependence; lack of NPPB block) were similar to the NaPi-1 induced P i -uptake, but no Ip could be recorded at P i -concentrations ≤3 mm. In summary, the present data suggest that Ip does not reflect charge transfer related to P i -uptake, but a P i -mediated modulation of GCl.
    Type of Medium: Electronic Resource
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