Electrogenic uptake of d-imino acids by luminal membrane vesicles from rabbit kidney proximal tubule

1989 ◽  
Vol 984 (2) ◽  
pp. 231-237 ◽  
Author(s):  
Hans Røigaard-Petersen ◽  
Christian Jacobsen ◽  
Henrik Jessen ◽  
Steen Mollerup ◽  
M. Iqbal Sheikh
1991 ◽  
Vol 260 (1) ◽  
pp. R186-R191 ◽  
Author(s):  
F. Van Der Goot ◽  
B. Corman

In the rabbit nephron, the luminal membrane surface area of the proximal convoluted tubule (PCT) is more than twice that of the proximal straight tubule (PST). What seemed to be an increase in histological specialization in solute and water transport is curiously reflected by a lower transepithelial water permeability per unit of apical membrane area in PCT than in PST. To evaluate what change in luminal membrane water permeability corresponds to this morphological difference, the osmotic permeabilities (Pf) of brush-border membrane vesicles isolated from PCT and PST of rabbit kidney were compared. D-Glucose uptake rates indicated proper separation of two populations of vesicles. Vesicle size measured by quasi-elastic light scattering was 123 +/- 7 nm and 125 +/- 6 nm for vesicles isolated from PCT and PST, respectively. Pf obtained by stop-flow light scattering techniques was of 106 +/- 6 microns/s in PCT vesicles and 191 +/- 7 microns/s in PST vesicles (T = 26 degrees C). In the presence of the sulfhydryl reagent HgCl2, the water permeabilities of both types of membrane dropped to comparable values. These data, which show an 80% increase in apical water permeability along the length of the proximal tubule, suggest that the number of proteic water channels per unit of membrane area is greater in PST than in PCT.


1987 ◽  
Vol 253 (1) ◽  
pp. F15-F20 ◽  
Author(s):  
H. Roigaard-Petersen ◽  
C. Jacobsen ◽  
M. Iqbal Sheikh

The mechanism of renal transport of L-proline by luminal-membrane vesicles isolated from proximal convoluted tubules of rabbit kidney was studied. It was found that H+ gradient (extravesicular greater than intravesicular) can drive the transport of L-proline into the vesicles both in the presence and absence of Na+. The stimulation of L-proline uptake by a pH gradient was additive with that produced by Na+. Saturation kinetic experiments revealed that pH gradient, in addition to Na+, increased the maximal uptake of L-proline by twofold. This is the first demonstration of H+-L-proline cotransport across luminal membrane of rabbit kidney proximal convoluted tubule. The physiological importance of this system is briefly discussed.


1992 ◽  
Vol 286 (1) ◽  
pp. 103-110 ◽  
Author(s):  
H Jessen ◽  
M I Sheikh

1. The mechanism of the renal transport of L-tryptophan by basolateral and luminal membrane vesicles prepared from either the pars convoluta or the pars recta of the rabbit proximal tubule was studied. The uptake of L-tryptophan by basolateral membrane vesicles from the pars convoluta was found to be an Na(+)-dependent transport event. The Na(+)-conditional influx of the amino acid was stimulated in the presence of an inwardly directed H+ gradient. Lowering the pH without an H+ gradient had no effect, indicating that L-tryptophan is co-transported with H+. 3. On the other hand, no transient accumulation of L-tryptophan was observed in the presence or absence of Na+ in basolateral membrane vesicles from the pars recta. 4. In luminal membrane vesicles from the pars recta, the transient Na(+)-dependent accumulation of L-tryptophan occurred via a dual transport system. In addition, an inwardly directed H+ gradient could drive the uphill transport of L-tryptophan into these vesicles in both the presence and the absence of an Na+ gradient. 5. By contrast, the uptake of L-tryptophan by luminal membrane vesicles from the pars convoluta was a strictly Na(+)-dependent and electrogenic transport process, mediated by a single transport component. 6. Investigation of the coupling ratio in luminal membrane vesicles suggested that 1 Na+:1 L-tryptophan are co-transported in the pars convoluta. In the pars recta, examination of the stoichiometry indicated that approx. 1 H+ and 2 Na+ (high affinity) or 1 Na+ (low affinity) are involved in the uptake of L-tryptophan.


1990 ◽  
Vol 272 (3) ◽  
pp. 839-842 ◽  
Author(s):  
J Blokkebak-Poulsen ◽  
M I Sheikh ◽  
C Jacobsen

The characteristics of 86Rb+ fluxes through conductive channels in basolateral-membrane vesicles isolated from pars recta of rabbit kidney proximal tubule were investigated. In RbCl-, KCl- and NaCl-loaded vesicles a transient and almost equal accumulation of 86Rb+ was observed. The uptakes of 86Rb+ were inhibited to the same extent by 10 mM-BaCl2 in all loadings. The accumulation was driven by an electrical diffusion potential. The 86Rb+ flux was dependent on intravesicular Ca2+. Increasing concentrations of Ca2+ gradually decreased the 86Rb+ uptake. At 10 microM-Ca2+ the radioisotope flux was below 20% of control. The vesicles containing the channel showed very low selectivity among the univalent cations K+, Rb+, Li+, Na+ and choline+.


1988 ◽  
Vol 254 (5) ◽  
pp. F628-F633
Author(s):  
H. Roigaard-Petersen ◽  
C. Jacobsen ◽  
M. I. Sheikh

The mechanism of renal transport of L-proline by luminal membrane vesicles prepared from proximal straight tubules (pars recta) of rabbit kidney was investigated. The following picture emerges from transport studies: an electrogenic and Na+-requiring system confined to this region of nephron exists for transport of L-proline with a high affinity (Km = 0.16 mM) and low capacity (Vmax = 3.5 nmol.mg protein-1.15 S-1). Lowering the pH from 7.5 to 5.5 increased the affinity (Km lowered from 0.16 mM at pH 7.5 to 0.08 mM at pH 5.5) without changing the maximal capacity of this system. Modification of histidyl residues of the intact luminal membrane vesicles by diethyl-pyrocarbonate (DEP) completely abolished the transient renal accumulation of L-proline. Simultaneous presence of Na+ and L-proline (10 mM) protects against DEP inactivation of renal transport of radioactive L-proline. We propose that a histidyl residue may be at or close to the active site of L-proline transporter in vesicles from the pars recta.


1990 ◽  
Vol 258 (2) ◽  
pp. F388-F396 ◽  
Author(s):  
H. Roigaard-Petersen ◽  
H. Jessen ◽  
S. Mollerup ◽  
K. E. Jorgensen ◽  
C. Jacobsen ◽  
...  

The characteristics of renal transport of glycine by luminal membrane vesicles isolated from either proximal convoluted part (pars convoluta) or proximal straight part (pars recta) of rabbit proximal tubule were investigated. In vesicles from pars convoluta two transport systems have been characterized: a Na(+)-dependent system with intermediate affinity (half-saturation 3.64 mM) and a Na(+)-independent system that, in the presence of H+ gradient (extravesicular greater than intravesicular), can accelerate the transport of glycine into these vesicles. This is the first demonstration of H(+)-glycine cotransport across the luminal membrane of rabbit kidney proximal convoluted tubule. By contrast, in membrane vesicles from pars recta, transport of glycine was strictly dependent on Na+ and occurred via a dual transport system, namely a high-affinity (half-saturation 0.34 mM) and a low-affinity system (half-saturation 8.56 mM). The demonstration of competition between the H(+)-gradient dependent uptake of glycine, L-alanine, and L-proline, but insignificant inhibition with L-phenylalanine in vesicles from pars convoluta suggests that glycine, L-proline, and L-alanine probably share a common proton gradient-dependent transport system. In vesicles from pars recta, the Na(+)-dependent uptake of glycine was inhibited by low concentrations of L-alanine and L-phenylalanine, whereas addition of L-proline to the incubation medium did not significantly alter the uptake of glycine, suggesting that the Na(+)-dependent high-affinity system for glycine located in pars recta is shared with the high-affinity L-alanine and L-phenylalanine but not L-proline transport system.


1994 ◽  
Vol 267 (5) ◽  
pp. F725-F736 ◽  
Author(s):  
S. Abdulnour-Nakhoul ◽  
R. N. Khuri ◽  
N. L. Nakhoul

The effect of norepinephrine (NE) on mechanisms of cellular Na+ transport in the isolated, perfused proximal tubule of Ambystoma tigrinum was examined. Single-barreled voltage and ion-selective microelectrodes were used to determine basolateral (V1), luminal (V2), and transepithelial (V3) membrane potentials and intracellular Na+ activity (alpha Nai). In CO2/HCO3- control solution, addition of NE (10(-6) M) to the bath caused depolarizations of V1, V2, and V3 are decreased alpha Nai. These effects were mimicked by isoproterenol and inhibited by propranolol. Addition of NE in the absence of luminal Na+ and substrates did not cause any changes in V1, V2, V3, or alpha Nai. NE did not affect the changes in membrane potential difference (PD) or alpha Nai caused by removal and readdition of luminal substrates and/or Na+. To study the effect of NE on Na-K-adenosinetriphosphatase (Na-K-ATPase), the pump was inhibited by external K+ removal and then reactivated by readdition of 12 mM K+ to the bath in the presence and absence of NE. Reactivation of the pump caused hyperpolarization of membrane PDs, and alpha Nai recovered monotonically in 3-5 min. The peak hyperpolarizations of V1 and V2 (approximately 1 min) were significantly larger in the presence of NE. During the first 3 min, and also at the same alpha Nai, the rate of decrease of alpha Nai was significantly faster in the presence of NE. In conclusion, these results show a direct effect of NE on cell membrane PDs and alpha Nai in the kidney proximal tubule. Most likely, beta-receptors are involved in mediating the action of NE. Neither Na/H exchange nor Na-substrate cotransport at the luminal membrane are affected by NE. On the other hand, NE activates Na-K-ATPase.


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