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Biomedical subjects

R Kinne

Publications and source records attributed to R Kinne.

At least 109 records · Page 6Linked to original sources
PubMed1977

Carbonic anhydrase activity of isolated brush border and basal-lateral membranes of renal tubular cells.

Carbonic anhydrase activity was found and characterized in isolated homogeneous preparations of microvilli and basal infoldings of the rat renal tubular cells. This membrane-bound activity was about 3% of the total carbonic anhydrase activity of the renal cortex. Compared to the renal cytoplasmic soluble form, the membrane-bound enzymes had the same Km for CO2 and were equally resistant to inhibition by chloride. However, they were less inhibited by different sulfonamides. The implication of the existence of membrane-bound enzymes in the renal tubular cells is discussed.

Animals↗
PubMed1977

[The function of kidney membranes].

Plasma membranes were isolated from rat kidney and their transport properties for sodium, calcium, protons, phosphate, glucose, lactate, and phenylalanine were investigated. The results demonstrate that the luminal plasma membrane, which is in contact with the primary urine, and the contraluminal plasma membrane, which is in contact with the interstitial fluid, differ in their content of ion-sensitive ATPases and sodium-cotransport systems. This allows conclusions on the mechanism of renal transport in which luminal and contraluminal membranes are arranged functionally in series.

Adenosine Triphosphatases↗
PubMed1976

Phosphate transport into brush-border membrane vesicles isolated from rat small intestine.

Uptake of Pi into brush-border membrane vesicles isolated from rat small intestine was investigated by a rapid filtration technique. The following results were obtained. 1. At pH 7.4 in the presence of a NaCl gradient across the membrane (sodium concentration in the medium higher than sodium concentration in the vesicles), phosphate was taken up by a saturable transport system, which was competitively inhibited by arsenate. Phosphate entered the same osmotically reactive space as D-glucose, which indicates that transport into the vesicles rather than binding to the membranes was determined. 2. The amount of phosphate taken up initially was increased about fourfold by lowering the pH from 7.4 to 6.0.3. When Na+ was replaced by K+, Rb+ or Cs+, the initial rate of uptake decreased at pH 7.4 but was not altered at pH 6.0.4. Experiments with different anions (SCN-,Cl-, SO42-) and with ionophores (valinomycin, monactin) showed that at pH 7.4 phosphate transport in the presence of a Na+ gradient is almost independent of the electrical potential across the vesicle membrane, whereas at pH 6.0 phosphate transport involves the transfer of negative charge. It is concluded that intestinal brush-border membranes contain a Na+/phosphate co-transport system, which catalyses under physiological conditions an electroneutral entry of Pi and Na+ into the intestinal epithelial cell. In contrast with the kidney, probably univalent phosphate and one Na+ ion instead of bivalent phosphate and two Na+ ions are transported together.

Animals↗
PubMed1976

Phenylalanine uptake in isolated renal brush border vesicles.

The uptake of L-phenylalanine into brush border microvilli vesicles and basolateral plasma membrane vesicles isolated from rat kidney cortex by differential centrifugation and free flow electrophoresis was investigated using filtration techniques. Brush border microvilli but not basolateral plasma membrane vesicles take up L-phenylalanine by an Na+-dependent, saturable transport system. The apparent affinity of the transport system for L-phenylalanine is 6.1 mM at 100 mM Na+ and for Na+ 13mM at 1 mM L-phenylalanine. Reduction of the Na+ concentration reduces the apparent affinity of the transport system for L-phenylalanine but does not alter the maximum velocity. In the presence of an electrochemical potential difference of Na+ across the membrane (etaNao greater than etaNai) the brush border microvilli accumulate transiently L-phenylalanine over the concentration in the incubation medium (overshoot pheomenon). This overshoot and the initial rate of uptake are markedly increased when the intravesicular space is rendered electrically more negative by membrane diffusion potentials induced by the use of highly permeant anions, of valinomycin in the presence of an outwardly directed K+ gradient and of carbonyl cyanide p-trifluoromethoxyphenylhydrazone in the presence of an outward-directed proton gradient. These results indicate that the entry of L-phenylalanine across the brush border membrane into the proximal tubular epithelial cells involves cotransport with Na+ and is dependent on the concentration difference of the amino acid, on the concentration difference of Na+ and on the electrical potential difference. The exit of L-phenylalanine across the basolateral plasma membranes is Na+-independent and probably involves facilitated diffusion.

Animals↗
PubMed1976

Phosphate transport by isolated renal brush border vesicles.

A sodium dependent specific transport system for phosphate is present in the brush border microvilli but absent from the basal-lateral plasma membranes. The apparent affinity of this transport system for phosphate is 0.08 mM at 100 mM sodium and pH 7.4. It is inhibited competitively by arsenate with an apparent inhibitor constant of 1.1 mM (100 mM sodium, pH 7.4). Sodium dependent phosphate uptake is two times higher at pH 8 compared to the uptake observed at pH 6. The apparent affinity of the transport system for sodium is also pH-dependent, half-maximal stimulation of uptake is found at pH 6 with 129 mM sodium, at pH 7.4 with 60 mM sodium and at pH 8 with 50 mM sodium. Under all conditions a nonhyperbolic dependence of phosphate uptake on the sodium concentration is observed. The uptake of phosphate by brush border microvilli vesicles shows a typical overshoot phenomenon in the presence of sodium gradient across the membrane (CNao greater than CNai). The amount of pohsphate taken up after 2 min is about twice the equilibrium value reached after 2 h of incubation. At pH 7.4 the initial rate of uptake is increased only slighyly (12%) by inside negative membrane diffusion potentials and inhibited to the same extent by inside positive membrane diffusion potentials. These results indicate that the entry of phosphate across the brush border membrane into the epithelial cell of the proximal tubule is coupled to the entry of sodium. The transfer of phosphate is dependent on its concentration gradient and on the concentration difference of sodium. The data are best explained by the following hypothesis: Both the primary phosphate as well as the secondary phosphate are transported in cotransport with sodium. The divalent form however seems to be transported preferentially. Its transport occurs electroneutral with 2 sodium ions; the monovalent phosphate also enters the cell together with 2 sodium ions but as a positively charged complex. The exit of phosphate across the contraluminal cell border is sodium independent and is favoured by the high intracellular phosphate concentration and the inside negative membrane potential.

Animals↗
PubMed1976

Sodium/proton antiport in brush-border-membrane vesicles isolated from rat small intestine and kidney.

Studies on proton and Na+ transport by isolated intestinal and renal brush-border-membrane vesicles were carried out to test for the presence of an Na+/H+-exchange system. Proton transport was evaluated as proton transfer from the intravesicular space to the incubation medium by monitoring pH changes in the membrane suspension induced by sudden addition of cations. Na+ transport was determined as Na+ uptake into the vesicles by filtration technique. A sudden addition of sodium salts (but not choline) to the membrane suspension provokes an acidification of the incubation medium which is abolished by the addition of 0.5% Triton X-100. Pretreatment of the membranes with Triton X-100 prevents the acidification. The acidification is also not observed if the [K+] and proton conductance of the membranes have been increased by the simultaneous addition of valinomycin and carbonyl cyanide p-trifluoromethoxyphenylhydrazone to the K+-rich incubation medium. Either valinomycin or carbonyl cyanide p-trifluoromethoxyphenylhydrazone when added alone do not alter the response of the membranes to the addition of Na+. Na+ uptake by brush-border microvilli is enhanced in the presence of a proton gradient directed from the intravesicular space to the incubation medium. Under these conditions a transient accumulation of Na+ inside the vesicles is observed. It is concluded that intestinal and renal brush-border membranes contain a NA+/H+ antiport system which catalyses an electroneutral exchange of Na+ against protons and consequently can produce a proton gradient in the presence of a concentration difference for Na+. This system might be involved in the active proton secretion of the small intestine and the proximal tubule of the kidney.

Animals↗
PubMed1976

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

Basal-lateral plasma membrane vesicles and brush border membrane vesicles were isolated from rat kidney cortex and the uptake of p-aminohippuric acid (PAH) into these vesicles was studied by Millipore filtration techniques. Both membrane preparations take up PAH into an osmotically reactive intravesicular space. The transport across the brush border membrane seems to involve only simple diffusion whereas in the basal-lateral plasma membrane in addition a specific transport system exists which is inhibited competitively by probenecid. The apparent affinity of this transport system for PAH is 5.4 X 10(-4) M and for probenecid 5.4 X 10(-5) M. PAH uptake into basal-lateral plasma membrane vesicles is influenced by alteration of the membrane potential. Maneuvers which render the intravesicular space more positive--as for example replacement of chloride by sulfate in the presence of a sodium gradient directed into the vesicles and addition of valinomycin in the presence of a potassium gradient directed into the vesicles--stimulate the uptake of PAH. Replacement of a sodium chloride gradient by a sodium thiocyanate gradient reduces the uptake probably by reducing the inside positive membrane potential. In the absence of salt gradients anion replacement and replacement of sodium by potassium does not affect PAH transport by basal-lateral plasma membranes. These results suggest that in isolated basal-lateral membranes transfer of PAH across the membrane is accompanied by a transfer of negative charge. They furthermore provide no evidence for the existence of a sodium-PAH cotransport system in this membrane preparation.

Aminohippuric Acids↗
PubMed1976

Polarity of proximal tubular epithelial cells in relation to transepithelial transport.

Transport properties of brush border microvilli and basal-lateral plasma membranes isolated from rat kidney cortex were studied by a millipore filtration technique. Brush border microvilli but not basal-lateral plasma membranes contain sodium dependent stereospecific transport system for D-glucose, L-phenylalanine and inorganic phosphate as indicated by saturability, countertransport and inhibition by structurally related compounds. Reduction of equilbrium uptake by increasing medium osmolarity suggests transport into an osmotically reactive space rather than binding to the membranes. Electrogenecity of the sodium-sugar and sodium-amino-acid cotransport system was established by their dependence on artificially imposed diffusion potentials. Also a NA+/H+ antiport system can be demonstrated in microvilli vesicles by demonstrating counterflow of both ions under short circuit conditions. Basal-lateral plasma membranes contain sodium independent stereospecific transport systems for sugars and amino acids. These results demonstrate a marked functional polarity of the cell membranes in respect to sodium dependent and sodium independent transport systems. This polarity in conjunction with the asymmetrical distribution of sodium between the intra- and extracellular space seems to enable the proximal tubule epithelial cells to perform active transepithelial transport.

Animals↗
PubMed1976

[Membrane function of the kidney].

Brush border membranes (luminal) and basal-lateral plasma membranes (contraluminal) of rat kidney proximal tubules were isolated by freeflow electrophoresis and their role in transepithelial transport was investigated. Enzymatic analysis revealed that the brush border membranes contain a bicarbonate stimulated ATPase and that the basal-lateral plasma membranes contain a Na+-K+-ATPase and a calcium stimulated ATPase. These findings suggest that an active, ATPase-mediated step in transepithelial bicarbonate or proton transport is located in the luminal membrane, whereas an active, ATPase-mediated step in transepithelial sodium and calcium transport is located in the contraluminal membrane. Transport studies with membrane vesicles demonstrated that sodium-dependent stereospecific transport systems for sugars, amino acids and phosphate are located in the brush border membrane; the basal-lateral plasma membranes contain sodium-independent transport systems for sugars; amino acids, phosphate and p-aminohippurate. The sodium-dependent systems represent sodium-substrate contransport systems which in the course of transepithelial transport derive energy from the transmembranel electrochemical potential difference of sodium for the intracellular accumulation and active transepithelial transport of sugars, amino acids and phosphate. The brush border membrane contains in addition a Na+/H+ exchange system which might be involved in the proton secretion of the proximal tubule. In the presence of a sodium gradient the permeability of the luminal membrane vesicles for L-lactate is higher than the permeability of the contraluminal membrane vesicles. This indicates that L-lactate-which is metabolized by the tubular epithelium-enters the tubular cell mainly from the tubular lumen. The role of membranes in the uptake of proteins by the tubular cell was investigated by isolation and biochemical characterization of microvilli, pinocytic vesicles and lysosomes. Pinocytic vesicles were found to be rich in acid phospholipids and glycoproteins which show a more rapid turnover than the proteins of the microvilli. It is concluded that pinocytic vesicles are biochemically defined entities with unique functions which are synthetized during the pinocytic process.

Adenosine Triphosphatases↗
PubMed1975

Distribution of parathyroid hormone-stimulated adenylate cyclase in plasma membranes of cells of the kidney cortex.

Free flow electrophoresis was employed to separate renal cortical plasma membranes into luminal (brush border microvilli) and contraluminal (basal-lateral membrane) fractions. During the separation adenylate cyclase activity was found to parallel the activity of Na+-K+-activated ATPase, an enzyme which is present in contraluminal but not in luminal membranes. In the basal-lateral membrane fraction the specific activities of adenylate cyclase and Na+-K+-activated ATPase were 4.4 and 4.6 times greater, respectively, than in the brush border fraction. The adenylate cyclase of the basal-lateral membrane fraction was specifically stimulated by parathyroid hormone which maximally increased enzyme activity eightfold. The biologically active (1-34) peptide fragment of paratyhroid hormone produced a 350% increase in adenylate cyclase activity. In contrast, calcitonin, epinephrine and vasopressin maximally stimulated the enzyme by only 55, 35 and 30%, respectively. These results indicate that adenylate cyclase, specifically stimulated by parathyroid hormone, is distributed preferentially in the contraluminal region of the plasma membrane of renal cortical epithelial cells.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1975

Distribution of membrane-bound cyclic AMP-dependent protein kinase in plasma membranes of cells of the kidney cortex.

Renal cortical plasms membranes were separated by free flow electrophoresis into luminal (brush border microvilli) and contraluminal (basal-lateral membrane) fractions. These membranes were found to contain an intrinsic, self-phosphorylating system which consists of a cyclic AMP-dependent protein kinase, a phosphorprotein phosphatase and the substrate(s) of these enzymes. The kinase, but not the phosphatase, was stimulated by cyclic AMP; maximal (1.7-fold) stimulation was effected at a cyclic AMP concentration of 0.1 muM. The degree of phosphorylation of the brush borders was six times greater than that of the basal-lateral membranes in the absence of cyclic AMP and 2.3-fold greater in the presence of cyclic AMP. This preferential phosphorylation of the luminal membrane by membrane-associated protein kinase(s) may play a role in the parathyroid hormone-mediated alterations of solute reabsorption in the proximal tubule.

Adenosine Triphosphatases↗
PubMed1975

Sugar transport by renal plasma membrane vesicles. Characterization of the systems in the brush-border microvilli and basal-lateral plasma membranes.

Uptake studies of D-and L-glucose were performed on vesicles derived from brush-border and basal-lateral membranes. The uptake of the sugars into the vesicles was osmotically sensitive and independent of glucose metabolism. In brush-border vesicles D-glucose but not L-glucose transport was Na-+-dependent, wn the presence of an initial Na+gradient. Basal-lateral membranes take up D-glucose faster than L-glucose, but the D-glucose uptake is significantly less sensitive to sodium removal and only moderately inhibited by phlorzin as compared to the prush-border fraction.

Animals↗