Hypotonicity increases basolateral taurine permeability in rabbit proximal convoluted tubule

1995 ◽  
Vol 268 (4) ◽  
pp. F595-F603 ◽  
Author(s):  
S. Breton ◽  
M. Marsolais ◽  
R. Laprade
Keyword(s):  
Initial Rate ◽  
Basolateral Membrane ◽  
Proximal Convoluted Tubule ◽  
Cell Swelling ◽  
Initial Increase ◽  
Volume Regulatory Decrease ◽  
Cellular Volume ◽  
Isotonic Condition ◽  
Rate Of Increase ◽  
A Cell

The permeabilities of the basolateral membrane of rabbit proximal convoluted tubule (PCT) to taurine (PTau) and glucose (PGlc) were estimated under control and hypotonic conditions using the initial rate of increase in cellular volume (CV) induced on isotonic replacement of 40 mM mannitol by one or the other of these substrates. Under control conditions, addition of taurine led to an increase in CV at an initial rate of 7.1 +/- 1.7%/min, leading to a cell swelling of 30.2 +/- 4.8% after 5 min (n = 6). Addition of glucose led to an increase in CV at an initial rate of 30.0 +/- 3.8%/min, leading to a cell swelling of 25.7 +/- 3.1% after 5 min (n = 7). After a period of recovery of 5 min in the absence of taurine or glucose, a 40 mosmol/kg hypotonic shock induced a cell swelling of 14.2 +/- 1.3 and 16.1 +/- 5.2%, respectively, followed by an almost complete volume regulatory decrease after 5 min. At that time, addition of taurine under continuous hypotonicity induced an increase in CV at an initial rate 2.57 +/- 0.17 times larger than that observed under the isotonic condition (P < 0.005), while addition of glucose induced an initial increase in CV identical to that observed under the isotonic condition. The increases in CV observed on addition of taurine were completely abolished in the absence of sodium under both isotonic and hypotonic conditions. The permeability to K+ was also estimated, in the absence of sodium, using the initial rate of increase in CV induced on isotonic replacement of 40 mM N-methyl-D-glucamine by K+.(ABSTRACT TRUNCATED AT 250 WORDS)

Volume regulation and intracellular calcium in the rabbit proximal convoluted tubule

1991 ◽  
Vol 260 (6) ◽  
pp. F861-F867 ◽  
Author(s):  
J. S. Beck ◽  
S. Breton ◽  
R. Laprade ◽  
G. Giebisch
Keyword(s):  
Intracellular Calcium ◽  
Cell Volume ◽  
Sodium Transport ◽  
Calcium Concentration ◽  
Basolateral Membrane ◽  
Potassium Conductance ◽  
Proximal Convoluted Tubule ◽  
Intracellular Calcium Concentration ◽  
Cell Swelling ◽  
Sustained Increase

The hypothesis that an increase of calcium leads to activation of calcium-activated ionic conductances during cell swelling was examined in the isolated perfused proximal convoluted tubule of the rabbit. Reduction of bath and luminal osmolality by 90 mosmol/kgH2O caused the cells to swell by 23.6 +/- 1.5% (n = 5) and intracellular calcium to rise from 227 +/- 35 to 347 +/- 60 nM (n = 6). Both these increases were transient, with volume decreasing to 5.5 +/- 1.2% above control and intracellular calcium concentration decreasing to 272 +/- 46 nM after 5-9 min. The addition of glucose and alanine to the tubule lumen to increase transcellular sodium transport caused a sustained increase in cell volume of 15.6 +/- 3.4% (n = 4). In parallel experiments, no significant increase in intracellular calcium concentration was observed. Addition of 1 microM of the calcium ionophore, ionomycin, reversibly increased intracellular calcium by 224 +/- 60 nM from a control value of 301 +/- 29 nM (n = 7) and reversibly depolarized the basolateral membrane by 3.6 +/- 0.9 mV (n = 5). However, there was no initial increase in the apparent transference number for potassium or chloride and no significant change in cell volume. We conclude from these observations that the sustained increase in basolateral potassium conductance observed when cells are swollen by hypotonicity or increased sodium transport (J. S. Beck and D. J. Potts. J. Physiol. Lond. 425: 369-378, 1990) is not due to a calcium-activated potassium conductance.

Relationship between sodium transport and intracellular ATP in isolated perfused rabbit proximal convoluted tubule

1991 ◽  
Vol 261 (4) ◽  
pp. F634-F639 ◽  
Author(s):  
J. S. Beck ◽  
S. Breton ◽  
H. Mairbaurl ◽  
R. Laprade ◽  
G. Giebisch
Keyword(s):  
Potassium Channels ◽  
Cell Volume ◽  
Intracellular Ph ◽  
Sodium Transport ◽  
Sodium Pump ◽  
Basolateral Membrane ◽  
Potassium Conductance ◽  
Transference Number ◽  
Proximal Convoluted Tubule ◽  
A Cell

The effect of alterations in sodium transport on cell ATP content and pH in the isolated perfused proximal convoluted tubule (PCT) of the rabbit was examined. Stimulating sodium transport by the addition of luminal glucose and alanine decreased cell ATP from 4.44 +/- 0.93 to 2.69 +/- 0.62 mM (n = 4), increased intracellular pH by 0.13 +/- 0.02 (n = 7), and increased cell volume by 0.10 +/- 0.02 nl/mm (n = 4). Blocking the sodium pump with 10(-4) M strophanthidin in tubules in which sodium transport had been stimulated increased cell ATP from 2.04 +/- 0.24 to 2.42 +/- 0.32 mM (n = 6). In parallel experiments the same dose of strophanthidin depolarized the basolateral membrane from -52.6 +/- 1.9 to -6.4 +/- 1.6 mV, depolarized the transepithelial potential from -3.2 +/- 0.3 to -0.1 +/- 0.1 mV, and reduced the basolateral membrane potassium transference number from 0.47 to 0.26 indicating a reduction in basolateral potassium conductance. Since strophanthidin caused a cell alkalinization of 0.15 +/- 0.03, this latter effect cannot be due to changes of intracellular pH. Strophanthidin caused no change in cell volume over the period studied, suggesting that stretch-activated potassium channels are not involved either. Instead, potassium conductance inhibition may be the result of the closure of ATP-sensitive potassium channels. These same channels might thus be partly responsible for the increase in potassium conductance commonly observed during stimulation of sodium transport.

Basolateral transport pathways for K+ and Cl- in rabbit proximal tubule: effects on cell volume

1991 ◽  
Vol 260 (1) ◽  
pp. F101-F109 ◽  
Author(s):  
L. Schild ◽  
P. S. Aronson ◽  
G. Giebisch
Keyword(s):  
Cell Volume ◽  
Basolateral Membrane ◽  
Cell Swelling ◽  
Volume Changes ◽  
Transport Pathways ◽  
Hypertonic Medium ◽  
Rate Of Increase ◽  
Kcl Transport ◽  
Cl Channels ◽  
Concentration Changes

To characterize the nature of K+ and Cl- transport pathways across basolateral membrane of rabbit proximal convoluted tubule, we used quantitative video microscopy to measure cell volume changes induced by rapid basolateral K+ and Cl- concentration changes. Elevating basolateral K+ resulted in cell swelling, which was largely inhibited by replacement of basolateral Cl- with cyclamate (85%) or by addition of 2 mM Ba2+ (72%). Substitution of basolateral Cl- by NO3- enhanced cell swelling, whereas substitution of Cl- by I- did not affect the K(+)-induced volume changes. Removal of Cl- from the bath reversed the cell swelling induced by raising K+ in the bath. Steady-state cell volume was 28% greater in hypotonic medium (250 mosmol/kgH2O) than in hypertonic medium (350 mosmol/kgH2O), and the rate of increase in cell volume induced by raising K+ was three times higher in hypotonic than in hypertonic medium. Substitution of Cl- by NO3- did not alter the effect of medium osmolality on K(+)-induced cell swelling, whereas addition of 0.2 mM diphenylamine-2-carboxylate inhibited the response (63%). We conclude that K(+)-induced cell swelling results from entry of K+ and Cl- into the cell across the basolateral membrane; it is proposed that transport of KCl across the basolateral cell membrane proceeds largely through two separate conductive pathways for K+ and Cl-. Cell swelling activates KCl transport occurring via K+ and Cl- channels across the basolateral membrane.

Evidence for the involvement of a K+ channel in isosmotic cell shrinking in vestibular dark cells

1992 ◽  
Vol 263 (3) ◽  
pp. C616-C622 ◽  
Author(s):  
P. Wangemann ◽  
N. Shiga ◽  
C. Welch ◽  
D. C. Marcus
Keyword(s):  
Initial Rate ◽  
Basolateral Membrane ◽  
Regulatory Volume Decrease ◽  
Cell Swelling ◽  
K Channel ◽  
Control Solution ◽  
Dark Cells ◽  
Physiological Relevance ◽  
K Secretion ◽  
Vestibular Dark Cells

Cell volume changes were measured in dark cells. Isosmotic addition of 21.4 mM K+, Rb+, Cs+, or NH4+ to a control solution containing 3.6 mM K+ caused piretanide-sensitive cell swelling (initial rate for K+, 0.100 +/- 0.005 microns/s; n = 119), suggesting dependence on the Na(+)-Cl(-)-K+ cotransporter. Subsequent isosmotic removal of 21.4 mM K+ caused piretanide-insensitive cell shrinking (initial rate, -0.104 +/- 0.005 microns/s; n = 119), which was inhibited by barium, lidocaine, quinidine, quinine, verapamil, and 4-aminopyridine but not tetraethylammonium (TEA) or glibenclamide, suggesting the involvement of K+ channel(s). Barium, lidocaine, quinine, quinidine, and 4-aminopyridine caused cell swelling in control solution (initial rate for barium, 0.011 +/- 0.004 microns/s; n = 6), suggesting that the K+ channel is also involved in efflux under control conditions. Cell shrinking was slowed by 21.4 mM extracellular K+, Rb+, or Cs+ but unaffected by Na+, Li+, TEA+, or NH4+ (all in the presence of piretanide and compared with N-methyl-D-glucamine), supporting the notion that the efflux mechanism is permeable to and/or inhibited by K+, Rb+, and Cs+. Cell shrinking was slowed by the presumed replacement of intracellular K+ by Cs+ but not by Rb+. Circumstantial evidence suggests that this putative K+ channel is present in the basolateral membrane. The physiological relevance of such a K+ channel might encompass regulatory volume decrease during K+ secretion.

scholarly journals Mechanism of basolateral membrane H+/OH-/HCO-3 transport in the rat proximal convoluted tubule. A sodium-coupled electrogenic process.

1985 ◽  
Vol 86 (5) ◽  
pp. 613-636 ◽  
Author(s):  
R J Alpern
Keyword(s):  
Initial Rate ◽  
Potassium Concentration ◽  
Basolateral Membrane ◽  
Chloride Concentration ◽  
Sodium Concentration ◽  
Proximal Convoluted Tubule ◽  
Extracellular Potassium ◽  
Sodium Removal ◽  
Cell Ph ◽  
Cell Depolarization

In order to examine the mechanism of basolateral membrane H+/OH-/HCO-3 transport, a method was developed for the measurement of cell pH in the vivo doubly microperfused rat proximal convoluted tubule. A pH-sensitive fluorescein derivative, (2',7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein, was loaded into cells and relative changes in fluorescence at two excitation wavelengths were followed. Calibration was accomplished using nigericin with high extracellular potassium concentrations. When luminal and peritubular fluids were pH 7.32, cell pH was 7.14 +/- 0.01. Decreasing peritubular pH from 7.32 to 6.63 caused cell pH to decrease from 7.16 +/- 0.02 to 6.90 +/- 0.03. This effect occurred at an initial rate of 2.4 +/- 0.3 pH units/min, and was inhibited by 0.5 mM SITS. Lowering the peritubular sodium concentration from 147 to 25 meq/liter caused cell pH to decrease from 7.20 +/- 0.03 to 6.99 +/- 0.01. The effect of peritubular sodium concentration on cell pH was inhibited by 0.5 mM SITS, but was unaffected by 1 mM amiloride. In addition, when peritubular pH was decreased in the total absence of luminal and peritubular sodium, the rate of cell acidification was 0.2 +/- 0.1 pH units/min, a greater than 90% decrease from that in the presence of sodium. Cell depolarization achieved by increasing the peritubular potassium concentration caused cell pH to increase, an effect that was blocked by peritubular barium or luminal and peritubular sodium removal. Lowering the peritubular chloride concentration from 128 to 0 meq/liter did not affect cell pH. These results suggest the existence of an electrogenic, sodium-coupled H+/OH-/HCO-3 transport mechanism on the basolateral membrane of the rat proximal convoluted tubule.

Inhibition of basolateral potassium conductance by taurine in the proximal convoluted tubule

1996 ◽  
Vol 271 (5) ◽  
pp. F1012-F1019 ◽  
Author(s):  
S. Breton ◽  
F. Belachgar ◽  
M. Marsolais ◽  
J. Y. Lapointe ◽  
R. Laprade
Keyword(s):  
Basolateral Membrane ◽  
Potassium Conductance ◽  
Membrane Conductance ◽  
Proximal Convoluted Tubule ◽  
Cell Swelling ◽  
Partial Recovery ◽  
Progressive Decrease ◽  
Electrophysiological Properties ◽  
The One ◽  
Dependent Transport

The effect of taurine on the electrophysiological properties of the basolateral membrane of the rabbit proximal convoluted tubule was examined. Short-duration isosmotic pulses of 40 mM taurine in the bath solution induced basolateral membrane depolarizations (delta Vbl) of 6.44 +/- 0.5 mV, which were reduced by 58% in absence of Na+. In presence of barium and quinine, delta Vbl values were reduced by 55% in Na(+)-containing bath solutions and were completely abolished in Na(+)-free solutions. Continuous addition of taurine into the bath solutions for a period of 5 min induced 1) a decrease in the partial conductance of the basolateral membrane to K+ (tK) from 0.39 to 0.23; 2) an increase in the partial conductance to Cl- (tcl) from 0.055 to 0.172; 3) a rapid and transient increase of the partial conductance mediated by the Na-HCO3 cotransporter (tNaHCO3) from 0.37 to 0.52, followed by a progressive decrease to 0.29; and 4) a depolarization of the basolateral membrane of 16 mV. The absolute membrane conductance mediated by the Na-HCO3 cotransporter was not initially affected by taurine, whereas that to K (GK) initially decreased by a maximal factor of 2 after 1 min, followed by a partial recovery after 5 min, and that to Cl (GCl) increased by a factor of 4. Addition of taurine after a hypotonicity-induced cell swelling produced an inhibition of GK comparable to the one observed under isotonic conditions. These results demonstrate the presence of an electrogenic Na-dependent transport of taurine and indicate that taurine inhibits GK.

Evidence against a proton pump in rabbit proximal convoluted tubule

1993 ◽  
Vol 264 (1) ◽  
pp. F175-F180 ◽  
Author(s):  
J. Beck ◽  
R. Laprade
Keyword(s):  
Basolateral Membrane ◽  
Proximal Convoluted Tubule ◽  
Proton Leak ◽  
Proton Extrusion ◽  
Cell Potential ◽  
Membrane Hyperpolarization ◽  
Initial Control ◽  
A Cell ◽  
Electrochemical Equilibrium

H+/OH- transport in the absence of bicarbonate was studied in the rabbit proximal convoluted tubule (PCT) perfused in vitro using measurements of membrane potential and intracellular pH (pHi). Blockade of apical Na/H exchange led to a cell acidification of 0.64 +/- 0.1 pH units from a control pHi of 7.27 +/- 0.04. A bafilomycin-insensitive recovery of pHi of 0.05 +/- 0.02 pH units occurred, but pHi did not exceed electrochemical equilibrium. A larger, sustained acidification of 0.87 +/- 0.07 from an initial control pHi of 7.25 +/- 0.05 induced by bilateral Na removal left pHi substantially below electrochemical equilibrium. These results suggest the absence of Na-independent active proton extrusion. We also examined the possibility that a passive electrogenic proton leak may exist. The removal of luminal glucose and alanine led to a basolateral membrane hyperpolarization of 31.3 +/- 0.5 mV, which was associated with a cell acidification of 0.15 +/- 0.02 pH units. These responses were reversed by addition of luminal glucose and alanine but not by depolarization by basolateral barium, suggesting that luminal glucose and alanine effects on pHi were due to changes other than cell potential. We conclude that, in the absence of bicarbonate, all active proton extrusion in the rabbit PCT is dependent on active Na transport and that a proton leak is negligible.

Luminal flow rate regulates proximal tubule H-HCO3 transporters

1992 ◽  
Vol 262 (1) ◽  
pp. F47-F54 ◽  
Author(s):  
P. A. Preisig
Keyword(s):  
Proximal Tubule ◽  
Flow Rate ◽  
Initial Rate ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Rate Dependence ◽  
Unstirred Layer ◽  
Disulfonic Acid ◽  
Luminal Flow

In vivo microperfusion was used to examine the mechanism of luminal flow rate dependence of proximal tubule acidification. Luminal flow rate was acutely changed between 5 and 40 nl/min, while luminal and peritubular capillary composition were held constant. With inhibition of basolateral membrane base transport by peritubular 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), cell pH (pHi) provides a sensitive index of apical membrane H secretory activity. At a luminal perfusate [HCO3] of 25 mM, progressive increases in luminal flow rate (5----15----25----40 nl/min) caused progressive increases in pHi. This effect was of a smaller magnitude with a luminal perfusate [HCO3] of 60 mM and was further decreased at a luminal perfusate [HCO3] of 100 mM. This pattern of diminished flow rate dependence at higher luminal [HCO3] is consistent with the presence of a luminal unstirred layer, whose composition can be modified by luminal flow rate. The activity of the apical membrane Na-H antiporter, assayed as the initial rate of pHi recovery from an acid load in the presence of peritubular DIDS, was faster at 40 compared with 5 nl/min. Basolateral membrane Na-3HCO3 symporter activity, assayed as the initial rate of pHi recovery from an alkali load in the absence of luminal and peritubular chloride, was faster at 40 compared with 5 nl/min. This effect was eliminated by luminal amiloride, suggesting an indirect effect of flow mediated by changes in pHi secondary to flow rate-dependent changes in apical membrane Na-H antiporter activity. In summary, increases in luminal flow rate directly increase apical membrane H secretion, possibly by modification of a luminal unstirred layer.(ABSTRACT TRUNCATED AT 250 WORDS)

Water permeability of apical and basolateral cell membranes of rat inner medullary collecting duct

1990 ◽  
Vol 259 (6) ◽  
pp. F986-F999 ◽  
Author(s):  
B. Flamion ◽  
K. R. Spring
Keyword(s):  
Water Flow ◽  
Water Permeability ◽  
Cell Membranes ◽  
Apical Membrane ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Volume Regulatory Decrease ◽  
Water Permeation ◽  
Inner Medullary Collecting Duct ◽  
Medullary Collecting Duct

To quantify the pathways for water permeation through the kidney medulla, knowledge of the water permeability (Posmol) of individual cell membranes in inner medullary collecting duct (IMCD) is required. Therefore IMCD segments from the inner two thirds of inner medulla of Sprague-Dawley rats were perfused in vitro using a setup devised for rapid bath and luminal fluid exchanges (half time, t1/2, of 55 and 41 ms). Differential interference contrast microscopy, coupled to video recording, was used to measure volume and approximate surface areas of single cells. Volume and volume-to-surface area ratio of IMCD cells were strongly correlated with their position along the inner medullary axis. Transmembrane water flow (Jv) was measured in response to a variety of osmotic gradients (delta II) presented on either basolateral or luminal side of the cells. The linear relation between Jv and delta II yielded the cell membrane Posmol, which was then corrected for membrane infoldings. Basolateral membrane Posmol was 126 +/- 3 microns/s. Apical membrane Posmol rose from a basal value of 26 +/- 3 microns/s to 99 +/- 5 microns/s in presence of antidiuretic hormone (ADH). Because of amplification of basolateral membrane, the ADH-stimulated apical membrane remained rate-limiting for transcellular osmotic water flow, and the IMCD cell did not swell significantly. Calculated transcellular Posmol, expressed in terms of smooth luminal surface, was 64 microns/s without ADH and 207 microns/s with ADH. IMCD cells in anisosmotic media displayed almost complete volume regulatory decrease but only partial volume regulatory increase.

Dynamic changes in viscoelastic properties in cytotoxic T-lymphocyte-mediated killing

1988 ◽  
Vol 91 (2) ◽  
pp. 179-189 ◽  
Author(s):  
K.L. Sung ◽  
L.A. Sung ◽  
M. Crimmins ◽  
S.J. Burakoff ◽  
S. Chien
Keyword(s):  
T Lymphocyte ◽  
Target Cell ◽  
Viscoelastic Properties ◽  
Cytotoxic T Lymphocyte ◽  
Cell Swelling ◽  
Initial Increase ◽  
Target Cells ◽  
Dynamic Changes ◽  
Lymphocyte Clone ◽  
Membrane Cell

The biophysical properties of cytotoxic T lymphocytes during the killing of their target cells was investigated by using a human cytotoxic T lymphocyte clone, F1, and the target cell, JY, for which it is specific. In single cytotoxic cell/target cell pairs after their conjugation there are changes in the viscoelastic properties of the target cell in association with the lethal hit delivery and post-binding cytolytic steps. On the basis of these changes in the target cell, the complex cytolytic event can be divided into stages: the viscoelastic coefficients exhibited an initial increase followed by a return to resting values; thereafter these coefficients decreased below control and then rose again prior to lysis. The eventual killing of the target cell involves bubbling and swelling of the nucleus, clustering of granules, damage to the cytoplasmic membrane, cell swelling, and lysis. The viscoelastic changes involved in target cell death suggest the loss of integrity of its cytoskeletal apparatus.

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