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  • 1
    Electronic Resource
    Electronic Resource
    Palo Alto, Calif. : Annual Reviews
    Annual Review of Physiology 63 (2001), S. 631-645 
    ISSN: 0066-4278
    Source: Annual Reviews Electronic Back Volume Collection 1932-2001ff
    Topics: Medicine , Biology
    Notes: Abstract Cl- transport in the loop of Henle is responsible for reclamation of 25-40% of the filtered NaCl load and for the formation of dilute urine. Our understanding of the physiologic and molecular mechanisms responsible for Cl- reabsorption in both the thin ascending limb and thick ascending limb of Henle's loop has increased greatly over the last decade. Plasma membrane Cl- channels are known to play an integral role in transcellular Cl- transport in both the thin and thick ascending limbs. This review focuses on the functional characteristics and molecular identities of these Cl- channels, as well as the role of these channels in the pathophysiology of disease.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 118 (1990), S. 269-278 
    ISSN: 1432-1424
    Keywords: Cl− channels ; bilayers ; PGO inhibition ; Cl− dependence ; open-time probability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary We evaluated the effects of vawrying aqueous Cl− concentrations, and of the arginyl- and lysyl-specific reagent phenylglyoxal (PGO), on the properties of Cl− channels fused from basolaterally enriched renal medullary vesicles into planar lipid bilayers. The major channel properties studied were the anion selectivity sequence, anionic requirements for, channel activity. and the efects of varying Cl− concentrations and/or PGO on the relation between holding voltageV H -mV) and open-time probability (P o). Reducingcis Cl− concentrations, in the range 50–320mm, produced a linear reduction in fractional open time (P v) with a half-maximal reduction inP o atcis Cl−≈170mM. Channel activity was sustained by equimolar replacement ofcis Cl− with F−, but not with impermeant isethionate. Fortrans solutions, the relation between Cl− concentration andP 0 at 10mm Cl−. Reducingcis Cl− had no effect on the gating charge (Z) for channel opening, but altered significantly the voltage-independent, energy (δG) for channel opening. Phenylglyoxal (PGO) reducedZ and altered δG for Cl− channel activity when added tocis, but nottrans solutions, Furthermore, in the presence ofcis PGO, reducing thecis Cl− concentration had no effect onZ but altered δG. Thus we propose thatcis PGO and,cis Cl− concentrations affect separate sites determining channel activity at the extracellular faces of, these Cl− channels.
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  • 3
    ISSN: 1432-1424
    Keywords: mTALH ; Cl− channel ; protein kinase A ; channel activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Cl− channels from basolaterally-enriched rabbit outer renal medullary membranes are activated either by increases in intracellular Cl− activity or by intracellular protein kinase A (PKA). Phosphorylation by PKA, however, is not obligatory for channel activity since channels can be activated by intracellular Cl− in the absence of PKA. The PKA requirement for activation of Cl− channels in certain secretory epithelia is, in contrast, obligatory. In the present studies, we examined the effects of PKA and intracellular Cl− concentrations on the properties of Cl− channels obtained either from basolaterally-enriched vesicles derived from highly purified suspensions of mouse medullary thick ascending limb (mTALH) segments, or from apical membrane vesicles obtained from two secretory epithelia, bovine trachea and rabbit small intestine. Our results indicate that the Cl− channels from mTALH suspensions were virtually identical to those previously described from rabbit outer renal medulla. In particular, an increase in intracellular (trans) Cl− concentration from 2 to 50 mm increased both channel activity (P o) and channel conductance (g Cl, pS). Likewise, trans PKA increased mTALH Cl− channel activity by increasing the activity of individual channels when the trans solutions were 2 mm Cl. Under the latter circumstance, PKA did not activate quiescent channels, nor did it affect g Cl. Moreover, when mTALH Cl− channels were inactivated by reducing cis Cl− concentrations to 50 mm, cis PKA addition did not affect P o. These results are consistent with the view that these Cl− channels originated from basolateral membranes of the mTALH. Cl− channels from apical vesicles from trachea and small intestine were completely insensitive to alterations in trans Cl− concentrations and demonstrated markedly different responses to PKA. In the absence of PKA, tracheal Cl− channels inactivated spontaneously after a mean time of 8 min; addition of PKA to trans solutions reactivated these channels. The intestinal Cl− channels did not inactivate with time. Trans PKA addition activated new channels with no effect on basal channel activity. Thus the regulation of Cl− channel activity by both intracellular Cl− and by PKA differ in basolateral mTALH Cl− channels compared to apical Cl− channels from either the tracheal or small intestine.
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  • 4
    ISSN: 1432-1424
    Keywords: Cl− channels/bilayers ; Cl− channels vesicles ; thick ascending limb ; channel conductance ; cAMP-dependent protein kinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary We examined the interactions of cAMP-dependent protein kinase and varying aqueous Cl− concentrations in modulating the activity of Cl− channels obtained by fusing basolaterally enriched renal outer medullary vesicles into planar lipid bilayers. Under the present experimental conditions, thecis andtrans solutions face the extracellular and intracellular aspects of these Cl− channels, respectively. Raising thetrans Cl− concentration from 2 to 50mm increased the channel open-time probability, raised the unit channel conductance, and affected the voltage-independent determinant (ΔG) of channel activity but not the gating charge (Winters, C.J., Reeves, W.B., Andreoli, T.E. 1990.J. Membrane Biol. 118:269–278). With 2mm trans KCl,trans addition of the catalytic subunit of PKA (C-PKA) plus ATP increased channel open-time probability and altered the voltage-independent determinant of channel activity without affecting either unit channel conductance or gating charge. The effect was ATP specific, did not occur with (C-PKA plus ATP) addition tocis solutions, and was abolished by denaturing C-PKA. Finally, (C-PKA plus ATP) activation of channel activity was not detected with relatively high (50mm)trans Cl− concentrations. These data indicate that (C-PKA plus ATP) might modulate Cl− channel activity by phosphorylation at or near the Cl−-sensitive site on the intracellular face of these channels.
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  • 5
    ISSN: 1432-1424
    Keywords: Cl− channels ; Cl−-interactive loci ; PGO ; TNBS ; Anion binding sites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A unique property of basolateral membrane Cl− channels from the mTAL is that the Cl− concentration facing the intracellular aspects of these channels is a determinant of channel open time probability (P 0 ). The K 1/2 for maximal activation of P 0 by Cl− facing intracellular domains of these channels is 10 mm Cl−. The present experiments evaluated the nature of these Cl−-interactive sites. First, we found that the impermeant anion isethionate, when exposed to intracellular Cl− channel faces, could augment P 0 with a K 1/2 in the range of 10 mm isethionate without affecting conductance (g Cl, pS). Second, pretreatment of the solutions facing the intracellular aspects of the channels with either 1 mm phenylglyoxal (PGO), an arginine-specific reagent, or the lysine/terminal amine reagent trinitrobenzene sulfonic acid (TNBS, 1 mm), prevented the activation of P 0 usually seen when the Cl− concentration of solutions facing intracellular channel domains was raised from 2 to 50 mm. However, when the Cl− channel activity was increased by first raising the Cl− concentration bathing intracellular channel faces from 2 to 50 mm, subsequent addition of either PGO or TNBS to solutions bathing intracellular Cl− channel faces had no effect on P 0 . We conclude that the intracellular aspects of these Cl− channels contain Cl−-interactive loci (termed [Cl] i ) which are accessible to impermeant anions in intracellular fluids and which contain arginineand lysine-rich domains which can be inactivated, at low ambient Cl− or isethionate concentrations, by interactions with PGO or TNBS.
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  • 6
  • 7
    Publication Date: 2001-03-01
    Description: ▪ Abstract  Cl− transport in the loop of Henle is responsible for reclamation of 25–40% of the filtered NaCl load and for the formation of dilute urine. Our understanding of the physiologic and molecular mechanisms responsible for Cl− reabsorption in both the thin ascending limb and thick ascending limb of Henle's loop has increased greatly over the last decade. Plasma membrane Cl− channels are known to play an integral role in transcellular Cl− transport in both the thin and thick ascending limbs. This review focuses on the functional characteristics and molecular identities of these Cl− channels, as well as the role of these channels in the pathophysiology of disease.
    Print ISSN: 0066-4278
    Electronic ISSN: 1545-1585
    Topics: Biology , Medicine
    Published by Annual Reviews
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