scholarly journals Basolateral membrane Na(+)-independent Cl-/HCO3- exchange in the inner stripe of the rabbit outer medullary collecting tubule.

1990 ◽  
Vol 95 (2) ◽  
pp. 347-367 ◽  
Author(s):  
S R Hays ◽  
R J Alpern
Keyword(s):  
Basolateral Membrane ◽  
Cell Voltage ◽  
Complete Removal ◽  
Distal Nephron ◽  
Cell Ph ◽  
Collecting Tubule ◽  
Nephron Segment ◽  
A Cell ◽  
Segment Cell

The inner stripe of the outer medullary collecting tubule is a major distal nephron segment in urinary acidification. To examine the mechanism of basolateral membrane H+/OH-/HCO3- transport in this segment, cell pH was measured microfluorometrically in the inner stripe of the rabbit outer medullary collecting tubule perfused in vitro using the pH-sensitive fluorescent dye, (2',7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein. Decreasing peritubular pH from 7.4 to 6.8 (changing [HCO3-] from 25 to 5 mM) caused a cell acidification of 0.25 +/- 0.02 pH units, while a similar luminal change resulted in a smaller cell acidification of only 0.04 +/- 0.01 pH units. Total replacement of peritubular Cl- with gluconate caused cell pH to increase by 0.18 +/- 0.04 pH units, an effect inhibited by 100 microM peritubular DIDS and independent of Na+. Direct coupling between Cl- and base was suggested by the continued presence of peritubular Cl- removal-induced cell alkalinization under the condition of a cell voltage clamp (K(+)-valinomycin). In addition, 90% of basolateral membrane H+/OH-/HCO3- permeability was inhibited by complete removal of luminal and peritubular Cl-. Peritubular Cl(-)-induced cell pH changes were inhibited two-thirds by removal of exogenous CO2/HCO3- from the system. The apparent Km for peritubular Cl- determined in the presence of 25 mM luminal and peritubular [HCO3-] was 113.5 +/- 14.8 mM. These results demonstrate that the basolateral membrane of the inner stripe of the outer medullary collecting tubule possesses a stilbene-sensitive Cl-/HCO3- exchanger which mediates 90% of basolateral membrane H+/OH-/HCO3- permeability and may be regulated by physiologic Cl- concentrations.

Changes in tubular basolateral membrane markers after chronic DOCA treatment

1983 ◽  
Vol 245 (1) ◽  
pp. F100-F109 ◽  
Author(s):  
G. El Mernissi ◽  
D. Chabardes ◽  
A. Doucet ◽  
A. Hus-Citharel ◽  
M. Imbert-Teboul ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Atpase Activity ◽  
Protein Content ◽  
Basolateral Membrane ◽  
Chronic Administration ◽  
Distal Nephron ◽  
Nephron Segments ◽  
Collecting Tubule ◽  
Nephron Segment ◽  
Cortical Collecting Tubule

Chronic administration of DOCA to rabbits is known to increase the surface area of the basolateral membrane and the Na-K-ATPase activity of the cortical collecting tubule (CCT). We attempted to ascertain 1) whether Na-K-ATPase is the only basolateral membrane marker induced by DOCA, and 2) whether CCT is the only nephron segment affected by this steroid. We measured the activity of Na-K-ATPase and adenylate cyclase (AC) and the protein content of nephron segments microdissected from control and DOCA-treated rabbits. Morphogenic effects of DOCA, assessed by 30-60% increases in protein content, were specifically observed in the distal convoluted tubule, CCT, and medullary collecting tubule. When expressed as a function of tubular length, Na-K-ATPase activity rose from 80 to 200% in all these segments, whereas the increments in AC of 40-70%, observed in response to four different hormones, occurred only in some of them. When expressed as a function of protein content, Na-K-ATPase activity increased but AC activity remained unchanged. This study indicates that the morphogenic action resulting from chronic DOCA administration affects the entire rabbit distal nephron. During this action Na-K-ATPase is the preferentially induced enzyme.

Effects of respiratory acidosis on HCO3- transport by rabbit collecting tubules

1988 ◽  
Vol 255 (4) ◽  
pp. F656-F665 ◽  
Author(s):  
T. D. McKinney ◽  
K. K. Davidson
Keyword(s):  
Calcium Concentration ◽  
Basolateral Membrane ◽  
Respiratory Acidosis ◽  
Bicarbonate Transport ◽  
Co2 Absorption ◽  
Distal Nephron ◽  
Outer Medulla ◽  
Collecting Tubules ◽  
Stimulation Of

These studies were performed to determine whether in vitro elevation of bath PCO2 with associated reduction in pH (acute respiratory acidosis) affected bicarbonate transport by isolated perfused rabbit cortical collecting tubules (CCT) and collecting tubules from the outer (OMCTos) and inner (OMCTis) stripes of the outer medulla. When the PCO2 was elevated and pH reduced from approximately 7.4 to 7.0 the rate of total CO2 absorption increased to 252% of that observed at pH 7.4 in CCT, 146% in OMCTos, and 150% in OMCTis. In OMCTis, pretreatment with colchicine inhibited the stimulation of total CO2 absorption associated with respiratory acidosis, whereas lumicolchicine did not. Similar inhibition was observed in the presence of maptam and a low calcium concentration and in the presence of a calmodulin inhibitor. No differences were observed in apical or basolateral membrane morphometry of principal or intercalated cells between control tubules and those subjected to respiratory acidosis. The results indicate that acute respiratory acidosis stimulates acidification by the rabbit distal nephron in vitro through a process(es) that, at least in OMCTis, evidently involves the cell cytoskeleton and changes in cell calcium and calmodulin activities.

Expression of claudin-7 and -8 along the mouse nephron

2004 ◽  
Vol 286 (6) ◽  
pp. F1063-F1071 ◽  
Author(s):  
Wing Y. Li ◽  
Catherine L. Huey ◽  
Alan S. L. Yu
Keyword(s):  
Tight Junction ◽  
Renal Tubule ◽  
Basolateral Membrane ◽  
Paracellular Permeability ◽  
Mouse Kidney ◽  
Integral Membrane Proteins ◽  
Distal Nephron ◽  
Nephron Segments ◽  
Nephron Segment

Claudins are integral membrane proteins of the tight junction that determine the magnitude and selectivity of paracellular permeability in epithelial tissues. The mammalian renal tubule exhibits considerable heterogeneity in the permeability properties of its different segments. To determine the nephron segment localization of claudin-7 and -8, immunofluorescence staining of mouse kidney sections was performed using isoform-specific antibodies. Claudin-8 was found to be expressed primarily at the tight junction along the entire aldosterone-sensitive distal nephron and in the late segments of the thin descending limbs of long-looped nephrons. This pattern of expression is consistent with the putative role of claudin-8 as a paracellular cation barrier. By contrast, claudin-7 was found in the same nephron segments as claudin-8, but it was expressed primarily at the basolateral membrane.

Role of basolateral carbonic anhydrase in proximal tubular fluid and bicarbonate absorption

2001 ◽  
Vol 280 (1) ◽  
pp. F146-F154 ◽  
Author(s):  
Shuichi Tsuruoka ◽  
Erik R. Swenson ◽  
Snezana Petrovic ◽  
Akio Fujimura ◽  
George J. Schwartz
Keyword(s):  
Carbonic Anhydrase ◽  
Basolateral Membrane ◽  
Bathing Solution ◽  
Fluid Absorption ◽  
Cell Ph ◽  
Before And After ◽  
Proximal Tubular ◽  
Membrane Bound

Membrane-bound carbonic anhydrase (CA) is critical to renal acidification. The role of CA activity on the basolateral membrane of the proximal tubule has not been defined clearly. To investigate this issue in microperfused rabbit proximal straight tubules in vitro, we measured fluid and HCO3 −absorption and cell pH before and after the extracellular CA inhibitor p-fluorobenzyl-aminobenzolamide was applied in the bath to inhibit only basolateral CA. This inhibitor was 1% as permeant as acetazolamide. Neutral dextran (2 g/dl, molecular mass 70,000) was used as a colloid to support fluid absorption because albumin could affect CO2 diffusion and rheogenic HCO3 − efflux. Indeed, dextran in the bath stimulated fluid absorption by 55% over albumin. Basolateral CA inhibition reduced fluid absorption (∼30%) and markedly decreased HCO3 − absorption (∼60%), both reversible when CA was added to the bathing solution. In the presence of luminal CA inhibition, which reduced fluid (∼16%) and HCO3 − (∼66%) absorption, inhibition of basolateral CA further decreased the absorption of fluid (to 74% of baseline) and HCO3 − (to 22% of baseline). CA inhibition also alkalinized cell pH by ∼0.2 units, suggesting the presence of an alkaline disequilibrium pH in the interspace, which would secondarily block HCO3 − exit from the cell and thereby decrease luminal proton secretion (HCO3 −absorption). These data clearly indicate that basolateral CA has an important role in mediating fluid and especially HCO3 −absorption in the proximal straight tubule.

scholarly journals Inhibition of Na(+)-independent H+ pump by Na(+)-induced changes in cell Ca2+.

1991 ◽  
Vol 98 (4) ◽  
pp. 791-813 ◽  
Author(s):  
S R Hays ◽  
R J Alpern
Keyword(s):  
Apical Membrane ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Chelating Agent ◽  
Cell Ph ◽  
Pump Activity ◽  
Acid Load ◽  
Medullary Collecting Duct ◽  
Induced Changes

Apical membrane H+ extrusion in the renal outer medullary collecting duct, inner stripe, is mediated by a Na(+)-independent H+ pump. To examine the regulation of this transporter, cell pH and cell Ca2+ were measured microfluorometrically in in vitro perfused tubules using 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein and fura-2, respectively. Apical membrane H+ pump activity, assayed as cell pH recovery from a series of acid loads (NH3/NH+4 prepulse) in the total absence of ambient Na+, initially occurred at a slow rate (0.06 +/- 0.02 pH units/min), which was not sufficient to account for physiologic rates of H+ extrusion. Over 15-20 min after the initial acid load, the rate of Na(+)-independent cell pH recovery increased to 0.63 +/- 0.09 pH units/min, associated with a steady-state cell pH greater than the initial pre-acid load cell pH. This pattern suggested an initial suppression followed by a delayed activation of the apical membrane H+ pump. Replacement of peritubular Na+ with choline or N-methyl-D-glucosamine resulted in an initial spike increase in cell Ca2+ followed by a sustained increase in cell Ca2+. The initial rate of Na(+)-independent cell pH recovery could be increased by elimination of the Na+ removal-induced sustained cell Ca2+ elevation by: (a) performing studies in the presence of 135 mM peritubular Na+ (1 mM peritubular amiloride used to inhibit basolateral membrane Na+/H+ antiport); (b) clamping cell Ca2+ low with dimethyl-BAPTA, an intracellular Ca2+ chelating agent; or (c) removal of extracellular Ca2+. Cell acidification induced a spike increase in cell Ca2+. The late acceleration of Na(+)-independent cell pH recovery was independent of Na+ removal and of the method used to acidify the cell, but was eliminated by prevention of the cell Ca2+ spike and markedly delayed by the microfilament-disrupting agent, cytochalasin B. This study demonstrates that peritubular Na+ removal results in a sustained elevation in cell Ca2+, which inhibits the apical membrane H+ pump. In addition, rapid cell acidification associated with a spike increase in cell Ca2+ leads to a delayed activation of the H+ pump. Thus, cell Ca2+ per se, or a Ca(2+)-activated pathway, can modulate H+ pump activity.

Cellular ultrastructure of Amphiuma distal nephron: effects of exposure to potassium

1984 ◽  
Vol 247 (3) ◽  
pp. C204-C216 ◽  
Author(s):  
B. Stanton ◽  
D. Biemesderfer ◽  
D. Stetson ◽  
M. Kashgarian ◽  
G. Giebisch
Keyword(s):  
Membrane Surface ◽  
Basolateral Membrane ◽  
Distal Tubule ◽  
Thick Ascending Limb ◽  
Distal Nephron ◽  
Structural Adaptation ◽  
Cellular Ultrastructure ◽  
Potassium Adaptation ◽  
Collecting Tubule ◽  
Collecting Tubules

The cellular ultrastructure of the renal distal nephron of the salamander, Amphiuma means, was examined by electron-microscopic and stereological techniques before and after exposure to potassium in the ambient environment. The distal nephron of Amphiuma is composed of three ultrastructurally distinct segments: early distal (or diluting segment), late distal, and collecting tubule. The early distal tubule structurally resembles the mammalian thick ascending limb of Henle's loop. Large renin-like granules are present in the smooth muscle cells of the afferent arteriole in the vicinity of the early distal tubule, suggesting the presence of a rudimentary juxtaglomerular apparatus. Late distal tubules are composed of one large cell type characterized by extensive basal membrane invaginations, often extending to the luminal membrane. Collecting tubules contain principal and intercalated cells that are ultrastructurally similar to cells of the mammalian cortical collecting tubule. Exposure to potassium had no effect on the ultrastructure of early distal cells but led to a sharp increase in the basolateral membrane surface density of principal cells in the collecting tubule (1.17 +/- 0.08-1.63 +/- 0.13 micron2/micron3). Potassium adaptation leads to a similar structural response in the mammalian collecting tubule. Since Amphiuma collecting tubules can be isolated and perfused in vitro and impaled with ion- and voltage-sensitive microelectrodes, the observed structural adaptation suggests that the collecting tubule may be a useful preparation to study the cellular mechanisms of potassium adaptation.

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.

A distal nephron glycoprotein that has different cell surface distributions on MDCK cell sublines

1987 ◽  
Vol 253 (3) ◽  
pp. C433-C443 ◽  
Author(s):  
G. K. Ojakian ◽  
R. E. Romain ◽  
R. E. Herz
Keyword(s):  
Cell Surface ◽  
Mdck Cell ◽  
Mdck Cells ◽  
Basolateral Membrane ◽  
Culture Collection ◽  
Rabbit Kidney ◽  
Distal Nephron ◽  
Surface Polarity ◽  
Nephron Segment ◽  
Collecting Tubules

Monoclonal antibodies that recognize three distinct epitopes of a 23-kDa glycoprotein (gp23) on the plasma membrane of MDCK cells were used to study cell-surface polarity. Immunofluorescence microscopy of MDCK cells obtained from the American Type Culture Collection demonstrated that gp23 was nonpolarized in approximately 50% of the cells (on both apical and basolateral membranes), whereas, in the remaining cells, gp23 had a polarized distribution (basolateral only). This heterogeneous gp23 cell-surface staining was not observed in a variety of other MDCK sublines. Instead, gp23 was found to have a nonpolarized distribution on MDCK cells that produced monolayers with low transepithelial electrical resistances (less than 220 omega X cm2) and was localized only to the basolateral membrane of MDCK cell lines capable of generating considerably higher transepithelial electrical resistances (770-2,220 omega X cm2). Immunofluorescence and immunoelectron microscopy of dog, rat, and rabbit kidney demonstrated that gp23 is a nephron segment-specific glycoprotein localized only to the distal and collecting tubules. These observations provide further evidence for the heterogeneity of the MDCK cell line. They also support a proposal that the origin of MDCK cells is the renal distal nephron.

scholarly journals Basolateral membrane Na/base cotransport is dependent on CO2/HCO3 in the proximal convoluted tubule.

1987 ◽  
Vol 90 (6) ◽  
pp. 833-853 ◽  
Author(s):  
R Krapf ◽  
R J Alpern ◽  
F C Rector ◽  
C A Berry
Keyword(s):  
Buffer Capacity ◽  
Basolateral Membrane ◽  
Proximal Convoluted Tubule ◽  
Proton Flux ◽  
Proton Fluxes ◽  
Cell Ph ◽  
Absolute Requirement ◽  
Ambient Co2

The mechanism of basolateral membrane base transport was examined in the in vitro microperfused rabbit proximal convoluted tubule (PCT) in the absence and presence of ambient CO2/HCO3- by means of the microfluorometric measurement of cell pH. The buffer capacity of the cells measured using rapid NH3 washout was 42.8 +/- 5.6 mmol.liter-1.pH unit-1 in the absence and 84.6 +/- 7.3 mmol.liter-1.pH unit-1 in the presence of CO2/HCO3-. In the presence of CO2/HCO3-, lowering peritubular pH from 7.4 to 6.8 acidified the cell by 0.30 pH units and lowering peritubular Na from 147 to 0 mM acidified the cell by 0.25 pH units. Both effects were inhibited by peritubular 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonate (SITS). In the absence of exogenous CO2/HCO3-, lowering peritubular pH from 7.4 to 6.8 acidified the cell by 0.25 pH units and lowering peritubular Na from 147 to 0 mM decreased cell pH by 0.20 pH units. Lowering bath pH from 7.4 to 6.8 induced a proton flux of 643 +/- 51 pmol.mm-1.min-1 in the presence of exogenous CO2/HCO3- and 223 +/- 27 pmol.mm-1.min-1 in its absence. Lowering bath Na from 147 to 0 mM induced proton fluxes of 596 +/- 77 pmol.mm-1.min-1 in its absence. The cell acidification induced by lowering bath pH or bath Na in the absence of CO2/HCO3- was inhibited by peritubular SITS or by acetazolamide, whereas peritubular amiloride had no effect. In the absence of exogenous CO2/HCO3-, cyanide blocked the cell acidification induced by bath Na removal, but was without effect in the presence of exogenous CO2/HCO3-. We reached the following conclusions. (a) The basolateral Na/base n greater than 1 cotransporter in the rabbit PCT has an absolute requirement for CO2/HCO3-. (b) In spite of this CO2 dependence, in the absence of exogenous CO2/HCO3-, metabolically produced CO2/HCO3- is sufficient to keep the transporter running at 30% of its control rate in the presence of ambient CO2/HCO3-. (c) There is no apparent amiloride-sensitive Na/H antiporter on the basolateral membrane of the rabbit PCT.

Ultrastructure of melanocyte-keratinocyte interactions and pigment transfer in vitro

1978 ◽  
Vol 36 (2) ◽  
pp. 650-651
Author(s):  
Raul I. Garcia ◽  
Evelyn A. Flynn ◽  
George Szabo
Keyword(s):  
Direct Injection ◽  
Skin Pigmentation ◽  
Freeze Fracture ◽  
Pigment Granule ◽  
Time Lapse ◽  
Ultrastructural Level ◽  
Lanthanum Tracer ◽  
A Cell

Skin pigmentation in mammals involves the interaction of epidermal melanocytes and keratinocytes in the structural and functional unit known as the Epidermal Melanin Unit. Melanocytes(M) synthesize melanin within specialized membrane-bound organelles, the melanosome or pigment granule. These are subsequently transferred by way of M dendrites to keratinocytes(K) by a mechanism still to be clearly defined. Three different, though not necessarily mutually exclusive, mechanisms of melanosome transfer have been proposed: cytophagocytosis by K of M dendrite tips containing melanosomes, direct injection of melanosomes into the K cytoplasm through a cell-to-cell pore or communicating channel formed by localized fusion of M and K cell membranes, release of melanosomes into the extracellular space(ECS) by exocytosis followed by K uptake using conventional phagocytosis. Variability in methods of transfer has been noted both in vivo and in vitro and there is evidence in support of each transfer mechanism. We Have previously studied M-K interactions in vitro using time-lapse cinemicrography and in vivo at the ultrastructural level using lanthanum tracer and freeze-fracture.

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