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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 138 (1994), S. 65-76 
    ISSN: 1432-1424
    Keywords: Cl- channel ; Na+ channel ; Ion fluxes ; Hormonal regulation ; Epithelial polarity ; cAMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The effect of antidiuretic hormone on transepithelial Na+ and Cl- transport and its modulation by aldosterone (10-6 m) was studied in the Xenopus laevis distal nephron cell line A6-C1 by measuring transepithelial electrophysiological parameters and bidirectional anion fluxes. Vasotocin (or vasopressin) induced a biphasic increase in transepithelial short-circuit current (I sc). Early and late effects were potentiated by aldosterone and could be mimicked by forskolin and BrcAMP, implicating cAMP as a mediator. The early increase in I sc (maximum 1–2 min after hormone addition) was resistant to 50 μm amiloride. Electrophysiological experiments with apical ion substitutions or basolateral bumetanide (0.5 mm), as well as flux studies with 125I- or 36C1-, indicated that this current represented Cl- secretion. The late increase in I sc appeared with a lag of 2–5 min and was maximal after 15–25 min. It corresponded to an increase in Na+ reabsorption, since it was amiloride sensitive. Bidirectional 36C1- flux measurements in aldosterone-treated monolayers maintained under open-circuit conditions showed that the large vasotocin-induced increase in Cl- permeability led, in these conditions, to a threefold increase of a baseline Cl- reabsorption. This study shows that vasotocin induces in A6-C1 cells both a rapid increase in Cl- permeability and a slower increase in Na+ transport. The Cl- permeability, which leads to Cl- secretion under short-circuit conditions, contributes, under the more physiological open-circuit conditions, to the transport of Na+ by allowing its co-reabsorption with Cl-.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 144 (1995), S. 93-110 
    ISSN: 1432-1424
    Keywords: Aldosterone ; Mineralocorticoid hormone ; Glucocorticoid hormone ; Hormonal regulation ; Na+ channel ; Na+,K+-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Conclusions The model for the adrenal steroid action on Na transport in tight epithelia as depicted in Fig. 3A and B dissociates two phases: an early phase during which the pre-existing Na transport machinery is activated and a late phase during which the transport capacity of the machinery is increased. These two sequential phases have been distinguished based on differences in functional aspects of the induced transport, on selective effects of agents interfering with transcriptional regulation and on a correlation of the late response phase with an increase in transport protein synthesis and expression [26, 45, 46, 98, 99, 124]. These observations suggest that a bimodal stimulation of Na transport could involve two different gene networks which are directly (in the physiological meaning) and independently stimulated by the action of the hormone-receptor complex and the following “molecular” cascades (see section Molecular and Physiological Cascades). The relatively clear temporal dissociation of the responses found in experimental situations is probably the consequence of inherent properties of the two networks. Indeed, to generate rapid functional changes, the genes involved in the early response must encode products which have relatively short half-lifes at the mRNA and protein levels. In contrast, the constitutive elements of the Na transport machinery that are increased during the late phase of adrenal steroid action have, as shown for the Na,K-ATPase [82], relatively long half-lifes. Consequently, even though changes in transcription may take place early in the course of the hormonal treatment, they impact on protein synthesis and pools only slowly and after a substantial lag period. On the one hand, ongoing research will soon provide more information on the nature, time course and hormone/receptor specificity of adrenal-steroid-regulated genes. On the other hand, the availability of new technical and molecular tools to study the proteins of the Na transport machinery greatly increases the possibilities for studying its regulation by adrenal steroids. Consequently, it will be a fascinating challenge to relate the data emerging from both approaches, and it appears that only a combination of methods and tools will allow to progressively fill the gap of understanding which still lies between the transcriptional effects and the transport regulation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 145 (1995), S. 193-204 
    ISSN: 1432-1424
    Keywords: Epithelial cell polarity ; Microfilament ; Microtubule ; Epithelial Na channel ; Cl channel ; Fluid phase endocytosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Antidiuretic hormone (ADH; 2.5 × 10−8 m vasotocin) produces a stimulation of apical fluid phase endocytosis, protein secretion and NaCl reabsorption in Xenopus laevis A6 distal nephron cell epithelia pretreated with aldosterone (10−6 m). The increase of NaCl transport is mediated by a sequential opening of apical Cl and Na conductances. The aim of this study was to characterize the actin and tubulin cytoskeleton of A6 cells and to assess the impact of its disruption on baseline and ADH-induced apical vesicular membrane movements and ion transport to test for possible functional links. The microfilament (MF) and microtubule (MT) networks and their disruption were visualized by confocal laser microscopy. Conditions of depolimerization were selected, by cytochalasin D or cold and nocodazole, respectively. MF disruption produced an increase in baseline apical protein secretion (exocytic movements) (plus 18%) and a decrease of its induction by ADH (minus 35%). MF disruption also increased baseline horseradish peroxidase uptake (endocytic movements) (plus 21%), however, without affecting its ADH-induced increase. In the case of MT disruption, the ADH-induced stimulation of both protein secretion and fluid phase endocytosis was decreased by 70 and 44%, respectively. At the ion transport level, MF and MT disruption only insignificantly affected the ADH-induced Cl conductance, while they decreased the ADH-induced stimulation of Na transport (amiloride-sensitive short-circuit current and conductance) by a factor of 2 to 4. In conclusion, both MT and MF disruption decrease ADH-induced apical protein secretion and Na conductance, while the ADH-induced apical Cl conductance is not significantly affected. Taken together the data support the hypothesis that the modulation of Na channel expression by apical vesicular membrane movements plays a role in Na transport expression and its regulation by ADH.
    Type of Medium: Electronic Resource
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