Bicarbonate reabsorption and electrophysiology of proximal tubules in uninephrectomized rats

1991 ◽  
Vol 81 (2) ◽  
pp. 141-146 ◽  
Author(s):  
Dirce M. Zanetta Limongi ◽  
Antonio Carlos Cassola ◽  
Viktoria Woronik ◽  
Gerhard Malnic
Keyword(s):  
Basolateral Membrane ◽  
Electrical Potential ◽  
Ringer Solution ◽  
Proximal Tubules ◽  
Potential Difference ◽  
Protonmotive Force ◽  
Electrical Potential Difference ◽  
Analogue Model ◽  
Significant Difference ◽  
Membrane Electrical Potential

1. The kinetics of acidification of luminal fluid in hypertrophied proximal tubules after unilateral nephrectomy was studied by stationary microperfusion and continuous measurement of luminal pH with antimony microelectrodes. 2. Trans-epithelial and basolateral membrane electrical potential differences were measured in order to detect modifications in electrogenic transport mechanisms under these conditions. 3. The values of stationary pH and HCO−3 concentration were significantly lower, the mean acidification half-time was not different and net reabsorptive HCO−3 fluxes in proximal tubules were significantly increased in uninephrectomized rats. According to an electrical analogue model, these results suggest (a) a reduction in the internal series resistance of the H+ pump, caused perhaps by an increased density of pump sites, and (b) an increase in the protonmotive force of the pump. 4. The trans-epithelial electrical potential difference measured in free flow conditions was significantly more lumen-positive in uninephrectomized rats. The trans-epithelial electrical potential difference measured during intraluminal perfusion with Ringer solution containing 30 mmol/l HCO−3 was significantly more negative in all groups studied. In uninephrectomized rats treated with acetazolamide, the trans-epithelial electrical potential difference was more lumen-negative than that in untreated uninephrectomized rats. These results are compatible with a steeper transepithelial Cl− gradient as well as with electrogenic, active H+ secretion. 5. There was no significant difference in the basolateral electrical potential difference between control and uninephrectomized rats. 6. In conclusion, our data show an increase in the transport rates of HCO−3 in the proximal tubule of uninephrectomized rats, which may be due to an increase in the density of transporters in the brush-border membrane, and an increased ability of the transport mechanism to create H+ gradients.

Cl- secretion by cultured shark rectal gland cells. II. Effects of forskolin on cellular electrophysiology

1991 ◽  
Vol 260 (4) ◽  
pp. C824-C831 ◽  
Author(s):  
W. M. Moran ◽  
J. D. Valentich
Keyword(s):  
Apical Membrane ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Ringer Solution ◽  
Rectal Gland ◽  
Electrical Potential Difference ◽  
Electrophysiological Properties ◽  
Shark Rectal Gland ◽  
Membrane Electrical Potential ◽  
Cl Conductance

Employing microelectrode techniques we have assessed the cellular electrophysiological properties of shark rectal gland (SRG) cells in primary culture. In the absence of secretagogues a 10-fold reduction in the Cl- concentration of the apical superfusate shark Ringer solution had little effect on either apical membrane electrical potential difference (Va) or fractional resistance (fRa), indicating little, if any, apical membrane Cl- conductance. Superfusing the basolateral surface with high-K+ shark Ringer solution (K+ increased 10-fold) depolarized the basolateral membrane electrical potential difference (Vb) by 43 mV, indicating that this barrier is largely K+ conductive. In addition, basolateral Ba2+ (5 mM) depolarized Vb by 12 mV and reduced fRa from 0.92 to 0.58, results consistent with a K(+)-conductive basolateral membrane in unstimulated SRG cells. Basolateral forskolin (10(-6) M) depolarized Va by 25 mV and caused a dramatic reduction in fRa from 0.97 to approximately 0.10. Under these conditions, a 10-fold decrease in apical superfusate Cl- concentration depolarized Va by 37 mV, revealing an adenosine 3',5'-cyclic monophosphate-induced apical membrane Cl- conductance. The time course of the forskolin-induced changes in Va and Vb suggests that the basolateral membrane K+ conductance increased and maintained the driving force for apical Cl- exit, as in other Cl(-)-secreting epithelia. These electrophysiological properties compare favorably with those of the perfused SRG tubule and indicate that SRG primary cultures are a suitable model for Cl(-)-secreting epithelia.

Potassium transport by flounder intestinal mucosa

1984 ◽  
Vol 246 (6) ◽  
pp. F946-F951 ◽  
Author(s):  
R. A. Frizzell ◽  
D. R. Halm ◽  
M. W. Musch ◽  
C. P. Stewart ◽  
M. Field
Keyword(s):  
Apical Membrane ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Electrical Potential Difference ◽  
Cell Type ◽  
K Secretion ◽  
Single Cell Type ◽  
Free Media ◽  
Membrane Electrical Potential ◽  
K Transport

We studied the mechanisms of K transport across an epithelium in which NaCl absorption is mediated primarily by Na/K/Cl cotransport at the apical membrane. Rubidium served as a reliable K substitute; under control conditions, both K and Rb were actively secreted. During secretion, K (Rb) enters across the basolateral membrane via the Na/K pump and exits across the apical membrane through K conductance pathways, since serosal ouabain or mucosal barium abolished K secretion, mucosal furosemide or Cl-free media blocked K secretion by interfering with access of Na to the pump, and elevated mucosal solution [K] or [Rb] depolarized the apical membrane electrical potential difference. Mucosal Ba unmasked active Rb absorption that could be blocked by mucosal furosemide. These findings illustrate active K absorption and secretion across an epithelium that comprises a single cell type in which opposing K fluxes across the apical membrane are mediated by Na/K/Cl cotransport entry and conductive K exit. The direction of transepithelial K transport is determined by the relative activities of these pathways.

Intracellular chloride activities in the isolated perfused shark rectal gland

1983 ◽  
Vol 245 (5) ◽  
pp. F640-F644
Author(s):  
M. J. Welsh ◽  
P. L. Smith ◽  
R. A. Frizzell
Keyword(s):  
Basolateral Membrane ◽  
Apical Cell ◽  
Electrical Potential ◽  
Electrochemical Potential ◽  
Potential Difference ◽  
Rectal Gland ◽  
Electrical Potential Difference ◽  
Apical Cell Membrane ◽  
Shark Rectal Gland ◽  
Electrochemical Equilibrium

The isolated, perfused shark rectal gland secretes Cl when stimulated with adenosine 3',5'-cyclic monophosphate (cAMP). To investigate the mechanism of secretion, we used Cl-selective and conventional (KCl-filled) microelectrodes to measure the intracellular Cl activity (aClc). Under nonsecreting conditions, the electrical potential difference across the basolateral membrane (psi b) was -78 m V and aClc was 57 mM, a value seven times greater than predicted for electrochemical equilibrium across the basolateral membrane. When theophylline and 8-bromo-cAMP were added to the perfusate, the transglandular electrical potential difference doubled and the rate of fluid secretion increased 20-fold; however, neither psi b nor aClc changed. During both nonsecreting and secreting conditions the intracellular accumulation of Cl results in an electrochemical potential difference favoring Cl exit across the apical cell membrane. The constancy of aClc despite the variation in secretion rate suggests that stimulation is associated with an equivalent enhancement of net Cl movement across both the apical and basolateral membranes. When stimulated glands were perfused with Na-free (choline) Ringer, secretion was abolished and aClc fell toward the value predicted for electrochemical equilibrium. These findings suggest that the "uphill" step in Cl secretion lies at the basolateral membrane, where cellular Cl accumulation probably involves secondary active transport; i.e., Cl entry is driven by an inwardly directed electrochemical potential difference for Na.

scholarly journals Electrical Potential Difference Measurements in Perfused Single Proximal Tubules of Necturus Kidney

1961 ◽  
Vol 44 (4) ◽  
pp. 679-687 ◽  
Author(s):  
Guillermo Whittembury ◽  
Erich E. Windhager
Keyword(s):  
Electrical Potential ◽  
Proximal Tubules ◽  
The Other ◽  
Potential Difference ◽  
Stopped Flow ◽  
Electrical Potential Difference ◽  
Necturus Maculosus ◽  
Other Hand ◽  
Glomerular Filtrate

Transtubular and peritubular face electrical potential differences (P.D.) of the proximal tubules of the kidney of the amphibian Necturus maculosus have been measured in situ. These measurements have been carried out both under normal conditions, when the tubular fluid originates in the glomerular filtrate, and under conditions when the composition of the tubular fluid has been altered using the stopped flow microperfusion technique. Under normal conditions the transtubular potential difference is 20 mv. (lumen-negative) and the P.D. across the peritubular face is 74 mv. (cell-negative). The P.D. across the luminal face is thus 54 mv. (cell-negative). This electrical asymmetry is not influenced by replacing the normal tubular fluid by NaCl, NaCl + mannitol, or by alteration in the intraluminal pH from 7 to 4. On the other hand, replacement of Na by K or choline and the addition of small amounts of DNP to the perfusate diminish this asymmetry.

Reversal of glibenclamide and voltage block of an epithelial KATP channel

1996 ◽  
Vol 271 (4) ◽  
pp. C1122-C1130 ◽  
Author(s):  
O. Mayorga-Wark ◽  
W. P. Dubinsky ◽  
S. G. Schultz
Keyword(s):  
Hydrophobic Interactions ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Membrane Vesicles ◽  
Voltage Gating ◽  
Electrical Potential Difference ◽  
Outer Solution ◽  
Channel Inhibition ◽  
K Concentration ◽  
Voltage Gate

K+ channels present in basolateral membrane vesicles isolated from Necturus maculosa small intestinal cells and reconstituted into planar phospholipid bilayers are inhibited by MgATP and sulfonylurea derivatives, such as tolbutamide and glibenclamide, when these agents are added to the solution bathing the inner mouth of the channel. In addition, these channels possess an intrinsic "voltage gate" and are blocked when the electrical potential difference across the channel is oriented so that the inner solution is electrically positive with respect to the outer solution. We now show that increasing the concentration of permeant ions such as K+ or Rb+ in the outer solution reverses channel inhibition resulting from the addition of 50 microM glibenclamide to the inner solution and also inhibits intrinsic voltage gating; these effects are not elicited by increasing the concentrations of the relatively impermeant ions, Na+ or choline, in the outer solution. Furthermore, increasing the K+ concentration in the outer solution in the absence of glibenclamide inhibits voltage gating, and, under these conditions, the subsequent addition of glibenclamide to the inner solution is ineffective. These results are consistent with a model in which the voltage gate is an open-channel blocker whose action is directly reversed by elevating the external concentration of relatively permeant cations and where the action of glibenclamide is to stabilize the inactivated state of the channel, possibly through hydrophobic interactions.

Active electrolyte transport in mammalian buccal mucosa

1988 ◽  
Vol 255 (3) ◽  
pp. G286-G291 ◽  
Author(s):  
R. C. Orlando ◽  
N. A. Tobey ◽  
V. J. Schreiner ◽  
R. D. Readling
Keyword(s):  
Buccal Mucosa ◽  
Short Circuit ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Short Circuit Current ◽  
Electrolyte Transport ◽  
Electrical Potential Difference ◽  
Ussing Chambers ◽  
Net Fluxes

The transmural electrical potential difference (PD) was measured in vivo across the buccal mucosa of humans and experimental animals. Mean PD was -31 +/- 2 mV in humans, -34 +/- 2 mV in dogs, -39 +/- 2 mV in rabbits, and -18 +/- 1 mV in hamsters. The mechanisms responsible for this PD were explored in Ussing chambers using dog buccal mucosa. After equilibration, mean PD was -16 +/- 2 mV, short-circuit current (Isc) was 15 +/- 1 microA/cm2, and resistance was 1,090 +/- 100 omega.cm2, the latter indicating an electrically "tight" tissue. Fluxes of [14C]mannitol, a marker of paracellular permeability, varied directly with tissue conductance. The net fluxes of 22Na and 36Cl were +0.21 +/- 0.05 and -0.04 +/- 0.02 mueq/h.cm2, respectively, but only the Na+ flux differed significantly from zero. Isc was reduced by luminal amiloride, serosal ouabain, or by reducing luminal Na+ below 20 mM. This indicated that the Isc was determined primarily by active Na+ absorption and that Na+ traverses the apical membrane at least partly through amiloride-sensitive channels and exits across the basolateral membrane through Na+-K+-ATPase activity. We conclude that buccal mucosa is capable of active electrolyte transport and that this capacity contributes to generation of the buccal PD in vivo.

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