Transport of p-aminohippuric acid by plasma membrane vesicles isolated from rat kidney cortex

1976 ◽  
Vol 361 (3) ◽  
pp. 269-277 ◽  
Author(s):  
W. Berner ◽  
R. Kinne
Keyword(s):  
Plasma Membrane ◽  
Rat Kidney ◽  
Membrane Vesicles ◽  
Kidney Cortex ◽  
Plasma Membrane Vesicles ◽  
Rat Kidney Cortex

scholarly journals Calcium ion transport across plasma membranes isolated from rat kidney cortex

1979 ◽  
Vol 178 (3) ◽  
pp. 549-557 ◽  
Author(s):  
P Gmaj ◽  
H Murer ◽  
R Kinne
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Rat Kidney ◽  
Membrane Vesicles ◽  
Free Flow ◽  
Kidney Cortex ◽  
Plasma Membrane Vesicles ◽  
Filtration Method ◽  
Rat Kidney Cortex ◽  
Calcium Ion Transport

Basal-lateral-plasma-membrane vesicles and brush-border-membrane vesicles were isolated from rat kidney cortex by differential centrifugation followed by free-flow-electrophoresis. Ca2+ uptake into these vesicles was investigated by a rapid filtration method. Both membranes show a considerable binding of Ca2+ to the vesicle interior, making the analysis of passive fluxes in uptake experiments difficult. Only the basal-lateral-plasma-membrane vesicles exhibit an ATP-dependent pump activity which can be distinguished from the activity in mitochondrial and endoplasmic reticulum by virtue of the different distribution during free-flow electrophoresis and its lack of sensitivity to oligomycin. The basal-lateral plasma membranes contain in addition a Na+/Ca2+-exchange system which mediates a probably rheogenic counter-transport of Ca2+ and Na+ across the basal cell border. The latter system is probably involved in the secondary active Na+-dependent and ouabain-inhibitable Ca2+ reabsorption in the proximal tubule, the ATP-driven system is probably more important for the maintenance of a low concentration of intracellular Ca2+.

scholarly journals Studies on the orientation of brush-border membrane vesicles

1978 ◽  
Vol 172 (1) ◽  
pp. 57-62 ◽  
Author(s):  
W Haase ◽  
A Schäfer ◽  
H Murer ◽  
R Kinne
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Rat Kidney ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Immunological Methods ◽  
Kidney Cortex ◽  
Asymmetric Distribution ◽  
Rat Kidney Cortex

Orientation of rat renal and intestinal brush-border membrane vesicles was studied with two independent methods: electron-microscopic freeze-fracture technique and immunological methods. With the freeze-fracture technique a distinct asymmetric distribution of particles on the two membrane fracture faces was demonstrated; this was used as a criterion for orientation of the isolated membrane vesicles. For the immunological approach the accessibility or inaccessibility of aminopeptidase M localized on the outer surface of the cell membrane to antibodies was used. With both methods we showed that the brush-border membrane vesicles isolated from rat kidney cortex and from rat small intestine for transport studies are predominantly orientated right-side out.

scholarly journals Immunoelectron Microscopic Localization of the Electrogenic Na/HCO3 Cotransporter in Rat and Ambystoma Kidney

2000 ◽  
Vol 11 (12) ◽  
pp. 2179-2189
Author(s):  
ARVID B. MAUNSBACH ◽  
HENRIK VORUM ◽  
TAE-HWAN KWON ◽  
SØREN NIELSEN ◽  
BRIAN SIMONSEN ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Proximal Tubule ◽  
Rat Kidney ◽  
Proximal Tubules ◽  
Kidney Cortex ◽  
Apical Plasma Membrane ◽  
Rat Kidney Cortex ◽  
C Terminus ◽  
Basolateral Plasma Membrane ◽  
Intracellular Vesicles

Abstract. Immunofluorescence analysis has revealed that electrogenic Na+/HCO3- (NBC1) is expressed in the proximal tubule of rat kidney and in the proximal and distal tubules of the salamander Ambystoma tigrinum kidney. The present study was undertaken to define the detailed subcellular localization of the NBC1 in rat and Ambystoma kidney using high-resolution immunoelectron microscopy. For this purpose, two rabbit polyclonal antibodies raised against amino acids 928 to 1035 and amino acids 1021 to 1035 of the C-terminus of rat kidney (rkNBC1) were developed. The affinity-purified antibodies revealed a strong band of approximately 140 kD in immunoblots of membranes from rat kidney cortex but no signal in membranes isolated from outer and inner medulla. Deglycosylation reduced the apparent molecular weight to approximately 120 kD, corresponding to the predicted molecular weight. A similar but weaker band was also present in membranes isolated from the lateral part of Ambystoma kidney. In rat kidney, immunohistochemistry confirmed the presence of rkNBC1 in convoluted segments of the proximal tubules. In ultrathin cryosections or Lowicryl HM20 sections from rat kidney cortex, distinct immunogold labeling was associated with the basolateral plasma membrane of segments S1 and S2 of proximal tubules, whereas in S3 no labeling was observed. The labeling density was similar at the basal and lateral plasma membrane and was specifically associated with the inner surface of the membrane consistent with the internal position of the C-terminus of the transporter. In contrast, rkNBC1 was absent from the apical plasma membrane and not observed in intracellular vesicles, including those closely associated with basolateral plasma membrane. In Ambystoma kidney, a weak labeling was present in the basolateral membrane of the proximal tubule and stronger labeling was observed in the late distal segment. The results demonstrate that rkNBC1 is expressed only in segment S1 and segment S2 of rat proximal tubule as well as Ambystoma proximal and late distal tubule and that rkNBC1 is present in both basal and lateral plasma membranes and absent in intracellular vesicles of the apical plasma membrane.

scholarly journals pH gradient as an additional driving force in the renal re-absorption of phosphate

1990 ◽  
Vol 271 (3) ◽  
pp. 687-692 ◽  
Author(s):  
J Strévey ◽  
S Giroux ◽  
R Béliveau
Keyword(s):  
Driving Force ◽  
Phosphate Uptake ◽  
Rat Kidney ◽  
Membrane Vesicles ◽  
Phosphate Transport ◽  
Kidney Cortex ◽  
Rat Kidney Cortex ◽  
Total Phosphate Concentration ◽  
Combined Action ◽  
Phosphate Carrier

The effects of the Na+ gradient and pH on phosphate uptake were studied in brush-border membrane vesicles isolated from rat kidney cortex. The initial rates of Na(+)-dependent phosphate uptake were measured at pH 6.5, 7.5 and 8.5 in the presence of sodium gluconate. At a constant total phosphate concentration, the transport values at pH 7.5 and 8.5 were similar, but at pH 6.5 the influx was 31% of that at pH 7.5. However, when the concentration of bivalent phosphate was kept constant at all three pH values, the effect of pH was less pronounced; at pH 6.5, phosphate influx was 73% of that measured at pH 7.5. The Na(+)-dependent phosphate uptake was also influenced by a transmembrane pH difference; an outwardly directed H+ gradient stimulated the uptake by 48%, whereas an inwardly directed H+ gradient inhibited the uptake by 15%. Phosphate on the trans (intravesicular) side stimulated the Na(+)-gradient-dependent phosphate transport by 59%, 93% and 49%, and the Na(+)-gradient-independent phosphate transport by 240%, 280% and 244%, at pH 6.5, 7.5 and 8.5 respectively. However, in both cases, at pH 6.5 the maximal stimulation was seen only when the concentration of bivalent trans phosphate was the same as at pH 7.5. In the absence of a Na+ gradient, but in the presence of Na+, an outwardly directed H+ gradient provided the driving force for the transient hyperaccumulation of phosphate. The rate of uptake was dependent on the magnitude of the H+ gradient. These results indicate that: (1) the bivalent form of phosphate is the form of phosphate recognized by the carrier on both sides of the membrane; (2) protons are both activators and allosteric modulators of the phosphate carrier; (3) the combined action of both the Na+ (out/in) and H+ (in/out) gradients on the phosphate carrier contribute to regulate efficiently the re-absorption of phosphate.

Inhibition by phenylglyoxal of the sodium-coupled fluxes of glucose and phosphate in renal brush-border membranes

1988 ◽  
Vol 66 (9) ◽  
pp. 1005-1012 ◽  
Author(s):  
R. Béliveau ◽  
M. Bernier ◽  
S. Giroux ◽  
D. Bates
Keyword(s):  
Brush Border ◽  
Direct Method ◽  
Rat Kidney ◽  
Membrane Vesicles ◽  
Kidney Cortex ◽  
Sodium Influx ◽  
Rat Kidney Cortex ◽  
Process Modification ◽  
Arginine Residues ◽  
Phosphate Influx

The coupling of phosphate and glucose transport to sodium in brush-border membrane vesicles from rat kidney cortex was studied after chemical modification of arginine residues by phenylglyoxal. Phosphate (10 mM) and sodium (20 mM) uptakes were linear for 6 s and stimulated in the presence of their cosubstrate. The sodium: phosphate stoichiometry measured by a direct method was 1.74. Sodium-independent phosphate and glucose influx were found to be unaffected by phenylglyoxylation. Phosphate- or glucose-independent sodium influx also remained unaltered by the treatment. However, phosphate influx measured with sodium was inhibited by 69% and sodium influx measured with phosphate was inhibited by 40%. When these values were corrected for uncoupled fluxes, the sodium influx coupled to phosphate and the phosphate influx coupled to sodium were inhibited by 93 and 95%, respectively. Glucose influx measured in the presence of sodium was inhibited by 36% and sodium influx in the presence of glucose was reduced by 39%. When the values were corrected for diffusion, these inhibitions were 95 and 100%, respectively. We conclude that the coupling of phosphate and glucose to sodium fluxes by the renal carriers requires the participation of arginine residue(s) in the translocation process. Modification of this arginine by phenylglyoxal leads to a marked inhibition of coupling. These results suggest the implication of arginine residues in the molecular coupling for both glucose and phosphate sodium symporters.

Kinetic model for phosphate transport in renal brush-border membranes

1988 ◽  
Vol 254 (3) ◽  
pp. F329-F336 ◽  
Author(s):  
R. Beliveau ◽  
J. Strevey
Keyword(s):  
Brush Border ◽  
Rat Kidney ◽  
Membrane Vesicles ◽  
Phosphate Transport ◽  
Kidney Cortex ◽  
Binary Complex ◽  
Rat Kidney Cortex ◽  
First Time ◽  
Phosphate Influx ◽  
Phosphate Carrier

Phosphate transport was studied in brush-border membrane vesicles purified from rat kidney cortex. Influx and efflux were strongly dependent on the presence of cis sodium; the rate of efflux, calculated by linear regression performed on the first time points, was much lower than the rate of influx (0.044 vs. 0.198 pmol.microgram protein-1.s-1). Trans phosphate had a stimulatory effect on phosphate influx (145% stimulation at 10 mM phosphate trans, with 0.2 mM phosphate cis). Trans phosphate was, however, inhibitory for phosphate efflux (89% inhibition at 10 mM phosphate trans). Trans effects of sodium were also studied. With 200 mM trans sodium, we observed 73% inhibition of phosphate influx and 60% inhibition of phosphate efflux. Studies involving sodium and phosphate present at the same time as trans substrates showed that the trans inhibition of phosphate influx by sodium could be completely reversed by trans phosphate. Trans inhibition of phosphate efflux by phosphate was not additive to the inhibition caused by sodium. Addition of trans phosphate had a stimulatory effect on sodium-independent influx, indicating that the binary complex (C-P) could translocate in efflux. These results indicate that the renal phosphate carrier presents a random binding scheme for the intra- and extravesicular sides of the membrane.

Indirect coupling to Na+ of p-aminohippuric acid uptake into rat renal basolateral membrane vesicles

1987 ◽  
Vol 253 (5) ◽  
pp. F795-F801 ◽  
Author(s):  
H. Shimada ◽  
B. Moewes ◽  
G. Burckhardt
Keyword(s):  
Transport System ◽  
Equilibrium Distribution ◽  
Rat Kidney ◽  
Basolateral Membrane ◽  
Membrane Vesicles ◽  
Kidney Cortex ◽  
Acid Uptake ◽  
Basolateral Membrane Vesicles ◽  
Rat Kidney Cortex ◽  
Indirect Coupling

Experiments with basolateral membrane vesicles prepared from rat kidney cortex were performed to study the mechanism by which p-aminohippuric acid (PAH) is taken up across the contraluminal membrane and is concentrated in proximal tubule cells. An inward Na+ gradient failed to stimulate [3H]PAH uptake compared with K+ or Li+ and did not cause intravesicular PAH accumulation above equilibrium distribution. In the absence of Na+, the dicarboxylates glutarate and suberate cis-inhibited and trans-stimulated [3H]PAH uptake, indicating a common transport system. In the presence of Na+, 10 microM glutarate in the incubation medium did not cis-inhibit, but rather stimulated [3H]PAH uptake and caused PAH accumulation above equilibrium distribution ("overshoot"). Li+ diminished this stimulation, but was without effect on [3H]PAH/PAH- and [3H]PAH/glutarate exchange. The data indicate the coexistence of a Na+ -coupled, Li+-sensitive transport system for dicarboxylates and a Li+ -insensitive PAH/dicarboxylate exchanger in the basolateral membrane. We propose that dicarboxylates are cotransported with Na+ into the cell and subsequently exchange for extracellular PAH at the basolateral membrane. PAH uptake is thereby indirectly coupled to Na+ via the Na+/dicarboxylate cotransporter.

scholarly journals Sulphate-ion/sodium-ion co-transport by brush-border membrane vesicles isolated from rat kidney cortex

1979 ◽  
Vol 182 (1) ◽  
pp. 223-229 ◽  
Author(s):  
Heinrich Lücke ◽  
Gertraud Stange ◽  
Heini Murer
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Rat Kidney ◽  
Electrical Potential ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Kidney Cortex ◽  
Electrical Potential Difference ◽  
Univalent Cations ◽  
Rat Kidney Cortex

Uptake of SO42− into brush-border membrane vesicles isolated from rat kindey cortex by a Ca2+-precipitation method was investigated by using a rapid-filtration technique. Uptake of SO42− by the vesicles was osmotically sensitive and represented transport into an intra-vesicular space. Transport of SO42− by brush-border membranes was stimulated in the presence of Na+, compared with the presence of K+ or other univalent cations. A typical ‘overshoot’ phenomenon was observed in the presence of an NaCl gradient (100mm-Na+ outside/zero mm-Na+ inside). Radioactive-SO42− exchange was faster in the presence of Na+ than in the presence of K+. Addition of gramicidin-D, an ionophore for univalent cations, decreased the Na+-gradient-driven SO42− uptake. SO42− uptake was only saturable in the presence of Na+. Counter-transport of Na+-dependent SO42− transport was shown with MoO42− and S2O32−, but not with PO42−. Changing the electrical potential difference across the vesicle membrane by establishing different diffusion potentials (anion replacement; K+ gradient±valinomycin) was not able to alter Na+-dependent SO42− uptake. The experiments indicate the presence of an electroneutral Na+/SO42−-co-transport system in brush-border membrane vesicles isolated from rat kidney cortex.

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