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A Kleinzeller

Publications and source records attributed to A Kleinzeller.

At least 37 records · Page 2Linked to original sources
PubMed1985

Trimethylamine oxide and the maintenance of volume of dogfish shark rectal gland cells.

Determinants of the steady-state vol of the dogfish shark (Squalus acanthias) rectal gland cells were studied. The cellular levels of trimethylamine oxide (TMAO) in fresh tissue and slices incubated aerobically 60 min in standard (TMAO-free) elasmobranch saline were close to those in the plasma (71 +/- 5 mM S.E.M.); therefore, under these conditions, the cell membrane appears to be impermeable to this solute. However, depolarization of the cells in high-K+ media produced a rapid loss of TMAO. Thus, TMAO is a major, effectively impermeant solute in the rectal gland cells. The osmolarity of cell solutes in tissue water (fresh and incubated slices) did not differ significantly from values in the plasma or incubation medium, demonstrating the absence of an osmotic pressure gradient across the cell membrane. An analysis of a simple model of cell solutes under steady-state conditions shows that the presence of an (effectively) impermeant osmolyte decreases the cellular concentration of bulk cations. The analysis is consistent with available observations on the distribution of cell Na+ and K+ in tissues containing high concentrations of (nitrogeneous) osmolytes. One simplifying assumption of the model, i.e., identity (or closeness) of the respective reflection coefficients sigma for Na+ and K+ passage through the cell membranes could not be verified. Compared to available data on the steady-state cellular fluxes of 42K+ in slices of the rectal gland, the uptake of 22Na+ by the tissue was slow (the derived rate constant k' = 0.017 min-1, i.e., about one tenth of that for K+).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Cell solute composition and potassium effects in slices of the rectal gland of the dogfish shark (Squalus acanthias).

Cell solute composition and apparent intracellular ionic concentrations (subscript i) were determined in rectal gland slices of the dogfish shark (Squalus acanthias). On aerobic incubation in elasmobranch Ringer's at 15 degrees C, the steady state values were: Ki+, 151 mM, close to its apparent electrochemical equilibrium; Cai2+, 1.7 mM; Mgi2+, 7.5 mM; Cl1-, 97 mM (maintained against its electrochemical potential gradient); the sum (Nai+ + Ki+) was about 66 meq/kg cell water lower than in the medium. Of free amino acids, only the effectively impermeable anionic taurine (32 mM) and glutamate (3.7 mM) significantly contribute to the Donnan system. Variations of the Ringer's K+ concentrations, or the presence of 0.5 mM ouabain, demonstrated the permeability of the cell membrane to na+, K+, and Cl-. Cell volume changes: 0.5 mM ouabain produced a gradual swelling and ionic changes consistent with an inhibition of the sodium pump. In high-K+ media the cells swelled massively and the membrane depolarized. A linear relationship between cell volume changes and the Nernst potential for K+ was found. These properties are consistent with a pump-and-leak system for cell volume maintenance. The K+ -induced cell swelling was found to be reversible on transfer of the tissue to regular Ringer's. The extrusion of cell water was associated with an influx of Na+ into, and an efflux of K+ from, the cells (both fluxes down-hill), as well as extrusion of Cl- against its concentration gradient.2+

Amino Acids↗
PubMed1985

Glucose transport in intestinal epithelia of winter flounder.

D-Glucose (Glc) transport was studied in stripped intestinal epithelial sheets of the winter flounder Pseudopleuronectes americanus. At 1 mM Glc and 15 degrees C, Glc uptake did not occur against a concentration gradient (free cell Glc 0.78 +/- 0.06 mM) but was inhibited by 0.5 mM phlorizin or ouabain. [U-14C]-Glc oxidation to 14CO2 was also depressed by these agents or the absence of Na+ and was saturable (Km 3.3 +/- 1.2 mM Glc; maximal velocity at saturating substrate concentration (Vmax) 4.0 +/- 2.0 mumol X g wet wt-1 X h-1]. No electrical transcellular manifestations of the Na-Glc cotransport system were seen in regular media. In the absence of Cl-, Glc and nonmetabolizable Glc analogs (alpha-methyl-D-glucopyranoside or 3-O-methyl-D-glucose) added mucosally elicited an increase in a serosally directed short-circuit current that was inhibitable by 0.5 mM mucosal phlorizin or 0.1 mM serosal ouabain and dependent on the external sugar concentration [Km 2.3 +/- 1.8 mM (range 0.6 to 6.1); Vmax 2.4 +/- 1.1 microA X cm-2 (range 0.8-4.2)]. Vmax for L-leucine transport was fivefold greater [13.4 +/- 7.3 microA X cm-2 (range 4.6-20.7)]. These results indicate the presence of a mucosal Na+-linked Glc absorptive system and reflect the paucity of transport sites for Glc relative to leucine.

Animals↗
PubMed1982

Anomalous patterns in cultured cell monolayers.

Gridlike patterns of differing cell density were observed in evenly seeded cell monolayers. Such patterns were obtained in five of six cell lines tested, suggesting widespread occurrence. The mechanism appears to involve small, transient temperature changes related to incubator tray structure. The very short time course of appearance of the patterns implicates attachment rather than growth as the critically affected factor. Impaired adhesion or directed sedimentation resulting from thermally induced microcurrents in the medium are the two most likely mechanisms.

Animals↗
PubMed1982

Metabolism of L-lactate by LLC-PK1 renal epithelia.

LLC-PK1 renal epithelial cells do not appreciably synthesize D-glucose or glycogen from L-lactate under the conditions described. [U-14C]L-lactate is metabolized to [14C]L-alanine and [14C]L-glutamate. Synthesis of alanine but not glutamate is decreased by the transaminase inhibitor, aminooxyacetate. LLC-PK1 cells also produce unlabeled urea in the presence of [14C]L-lactate. Evidence for net gluconeogenesis has thus not been obtained for this cell line. This is interesting because the LLC-PK1 cells possess an active sugar transport system, and both active sugar transport and gluconeogenesis are localized together in vivo in the proximal tubule.

Animals↗
PubMed1982

Renal sugar transport in the winter flounder. VI. Reabsorption of D-mannose.

The transport of D-mannose (Man) in flounder kidney was studied using renal clearance techniques in vivo and brush border membrane (BBM) vesicles in vitro. At plasma concentrations of 50-100 microM Man, the winter flounder (Pseudopleuronectes americanus) reabsorbed up to 70% of the filtered sugar. Man phosphates, but not free Man, accumulated in renal cells. Reabsorption of Man was reduced by phlorizin, D-glucose, methyl-alpha-D-glucoside, methyl-alpha-D-mannoside, and 2-deoxy-D-glucose. In BBM vesicles a Na+-dependent, phlorizin-sensitive overshoot in Man uptake (10 microM) was seen. Na+-dependent Man uptake was saturable, with an apparent Km of 127 microM. The transport properties for Man were identical in BBM vesicles from the winter flounder and southern flounder (Paralichthys lethostigma). The transport specificity was determined by cis-inhibition and trans-stimulation experiments with BBM. Glucose, galactose, 1,5-anhydro-D-mannitol (i.e., 1-deoxymannose), 2-deoxy-2-fluoro-D-glucose, and methyl-alpha-mannoside were shown to share the carrier-mediating mannose transport. 2-Deoxyglucose, methyl-alpha-2-deoxy-D-glucoside, and both the isomers (alpha and beta) of methyl-D-glucoside did not. In contrast, alpha-methyl-D-glucoside inhibited D-glucose transport both in vivo and in BBM vesicles. It is concluded that Man reabsorption in the flounder occurs via a Na+-cotransport system that also handles glucose but that differs from the glucose/methyl-alpha-D-glucoside reabsorptive pathway in that 1) an oxygen on C-1 is not required, and 2) an axial configuration for -OH on C-2 (C1 conformation) is readily accommodated.

Animals↗
PubMed1981

Transport of glucose analogues in rat lung.

Uptake of nonmetabolizable glucose analogues was investigated in isolated rat lungs ventilated with 5% CO2 in air and perfused with Krebs-Ringer bicarbonate medium. In some experiments, [5-3H]glucose, methyl(alpha-D-[U-14C]gluco)pyranoside (alpha-MG), 3-O-methyl-D-[U-14C]glucose (3-O-MG), or various inhibitors were added to the initial perfusate to determine glucose utilization by the rate of 3H2O production or to characterize the uptake of glucose analogues. [1,2-3H]polyethylene glycol (PEG) was used as an indicator of the extracellular water space and gave a mean value of 0.35 ml/g tissue; calculated mean intracellular H2O space was 0.48 ml/g tissue. Glucose utilization was 56.4 +/- 6.6 (mean +/- SE, n = 6) mumol . g dry wt-1 . h-1 and decreased by 61% with 3 mM phlorizin. After 1-2 h perfusion, intracellular alpha-MG concentration was 1.4-1.9 times the extracellular concentration. The mean tissue-to-medium ratio (T/M) for alpha-MG decreased by more than 30% in the presence of glucose (5.0 mM), phlorizin (0.5 mM), ouabain (0.5 mM), or the absence of external Na+. Intracellular 3-O-MG concentration did not exceed extracellular concentration during 2 h of perfusion and the mean T/M did not change with any of the inhibitors studied. The results indicate that the nonmetabolizable glucose analogue alpha-MG is accumulated against a concentration gradient by an active Na+-dependent transport process, whereas 3-O-MG is apparently taken up by a different mechanism.

3-O-Methylglucose↗
PubMed1981

Sugar absorption and secretion by winter flounder intestine.

Transport of sugars by winter flounder (Pseudopleuronectes americanus) intestine has been examined, and the following observations have been made. 1) No net absorption of D-galactose was found in controls; however, mucosally applied 0.1 mM phlorizin stimulated the mucosal-serosal flux with net sugar secretion. 2) Net absorption of methyl glycosides was inhibited by D-galactose, 0.1 mM ouabain, and phlorizin (without induction of secretion). 3) Net secretion of 2-deoxy-D-galactose was found without cellular accumulation of free sugar. The secretory process had a Km of 10 mM and was inhibited by serosally applied 0.1 mM phloretin, bilateral replacement of Ringer Na+ by choline, serosal ouabain, and D-galactose. 4) No net absorption or secretion of 2-deoxy-D-glucose was found with a variety of experimental conditions. 5) At least two active transport systems for sugars appear to be operative in the flounder intestine: a conventional Na+-dependent sugar transport system across the brush border that allows net absorption and a Na+-dependent secretory system that apparently also operates across the mucosal border. Because D-galactose is transported by both systems, there is no net flux of this sugar. Several possible models for the net secretory system are discussed.

Animals↗
PubMed1980

Active renal hexose transport. Structural requirements.

The active transport of methyl beta-D-galactoside and some other analogs of D-glucose and D-galactose was studied in slices of rabbit and renal cortex. 1. The non-metabolizable methyl beta-D-galactoside accumulates in renal cortical cells against its concentration gradient. At 1 mM substrate concentration (O2, 35 degrees C, 60 min incubation) the gradient was 2.36 +/- 0.11 S.E. (n = 33). The Kt was 1.50 +/- 0.02 mM. The active transport of the substrate was inhibited by dinitrophenol, phlorizin, absence of Na+ and by ouabain. This inhibition was incomplete, suggesting that the sugar may enter the cells by two separate pathways, only one of which was coupled to the down-hill electrochemical Na gradient. 2. The structural requirements for the interaction between substrate and the carrier were defined: (a) by testing the transport behavior of some analogs (1,5-anhydro-D-glucitol; methyl beta-thio-D-galadtoside; 3-deoxy-D-glucose; 4-deoxy-D-glucose; 5-thio-D-glucose; 6-deoxy-D-glucose and methyl-alpha-6-deoxy-D-glucoside); and (b) by inhibition analysis of methyl beta-D-galactoside transport. The role of each hydroxyl of the sugar molecule was tested by using a total of 41 analogs modified on each C by replacing -OH by -H, -O-CH3, -F and in some instances also by -SH. 3. The carrier is shared by D-glucose, D-galactose and their methyl glycosides. A pyranose ring is mandatory. The D-glucoconfiguration is preferred for the interaction with the carrier. 4. Replacement of -OH by -H or -S practically abolished (on C1, C2, C3) or greatly reduced (on C4) the affinity of the analog for the carrier. This was also confirmed by demonstrating that 1-deoxy-, and 3-deoxy-glucose and the thio-galactoside were not actively transported and their entry into the cells was not markedly affected by phlorizin, dinitrophenol, ouabain or absence of Na+. 4-Deoxy-D-glucose was taken up and its transport was inhibited by agents affecting the transport of methyl beta-D-galactoside. 5. Replacement of -OH by -F did not abolish the affinity of the analogs for the carrier, indicating hydrogen bonding between the carrier and the oxygens at C1, C2, C3, and C4. 6. 5-Thio-D-glucose was not transported against its concentration gradient and also poorly interacted with the carrier as shown by inhibition analysis. Hydrogen bonding between the carrier and the pyranose ring oxygen is suggested. 7. 6-Deoxyglucose is a potent inhibitor of methyl beta-D-galactoside transport although it is not actively taken up by the tissue. It is concluded that a hydroxyl at C6 is required for transport, but is not mandatory for an interaction with the carrier. However, 6-deoxy-D-galactose was ineffective as inhibitor. 8. The specificity of the carrier involved in the renal active transport of D-glucose, D-galactose and their methyl glycosides resembles qualitatively, and mostly also quantitatively that described for intestinal transport of these sugars.

Animals↗
PubMed1980

Sugar transport in the LLC-PK1 renal epithelial cell line: similarity to mammalian kidney and the influence of cell density.

Cells of confluent cultures of the established pig renal epithelial line, LLC-PK1, accumulate alpha-methyl-D-glucoside against a concentration gradient. This transport system is strongly inhibited by phlorizin and 6-deoxy-D-glucose, moderately inhibited by phloretin, and only weakly inhibited by 3-0-methyl-D-glucose, paralleling the situation in mammalian kidney. The time courses for the uptake of alpha-methyl-D-glucoside and for the carrier-mediated but passive uptake of 3-0-methyl-D-glucose are identical to those seen in mammalian kidney. Subconfluent cultures of LLC-PK1 cells are unable to accumulate alpha-methyl-D-glucoside, and their transport of this glucose analog is less sensitive to phlorizin inhibition than is the transport system in confluent cultures. Transmission electron micrographs show that cells from subconfluent cultures lack the microvillous surface seen in cells from confluent cultures. Cell density is thus a factor in the occurrence of structural and functional differentiated properties related to transport in these cells.

Animals↗
PubMed1980

Effects of ouabain and ortho vanadate on transport-related properties of the LLC-PK1 renal epithelial cell line.

Uptake of alpha-methyl-D-glucoside (AMG) by LLC-PK1 cells is inhibited by the uncoupler p-trifluoro-methoxyphenyl-hydrazone (FCCP) and by the absence of extracellular Na+, indicating that the transport system is energy- and Na+-dependent. We have previously demonstrated that transport of AMG by LLC-PK1 cells proceeds against a concentration gradient and is phlorizin-sensitive (Mullin et al., '80). Uptake of AMG was also inhibited by ouabain (OUA) but not by ortho-vanadate (VAN). Rubidium uptake also was affected by OUA but not by VAN. VAN, however, caused collapse of the three-dimensional domes of confluent LLC-PK1 monolayers much more rapidly and thoroughly than OUA. Since domes are presumably dependent upon the Na+ pump, yet VAN is not acting on transport-related functions of the OUA-sensitive (Na+ + K+)-ATPase, we hypothesize a direct effect of VAN on the water permeability of these cells. We also suggest that OUA does not act on these cells until domes collapse in normal course, and access of the OUA to the extracellular surface of antiluminal membranes is then achieved.

Animals↗
PubMed1978

Is hexokinase present in the basal lateral membranes of rat kidney proximal tubular epithelial cells?

The possible presence of hexokinase in basal lateral membranes from rat kidney proximal tubules was investigated. Basal lateral membranes were obtained from homogenates of rat kidney cortex by differential centrifugation and free flow electrophoresis. They were further purified by density gradient centrifugation. Hexokinase activity was measured as the phosphorylation of D-[U14C]glucose. Throughout the purification of the membranes, the specific activity of hexokinase decreased while that of (Na+ + K+)-ATPase increased. Hexokinase activity in all fractions could be quantitatively accounted for in terms of cytosolic and mitochondrial enzyme contributions. It is concluded that there is no hexokinase activity in basal lateral membranes from rat kidney.

Adenosine Triphosphatases↗
PubMed1978

Renal sugar transport in the winter flounder: V. secretion of 2-deoxy-D-galactose.

Isolated renal tubules and renal clearance techniques were used to characterize the renal handling of 2-deoxy-D-galactose (2-d-Gal) by the winter flounder (Pseudopleuronectes americanus). In vitro, energy-dependent, pH-sensitive uptake of 2-d-Gal (2-100 micron) was seen at the antiluminal face of the cell. Clearance measurements showed net secretion of 2-d-Gal in vivo. The mean clearance of 2-d-Gal in 18 fish was 0.98 +/- 0.16 ml/h while the glomerular filtration rate (GFR) was only 0.37 +/- 0.10 ml/h. Secretion was associated with marked renal accumulation of both 2-d-Gal and phosphorylated derivatives (2-d-Gal-1-phosphate). Tissue-to-plasma ratios (T/P) averaged 19 for free sugar and 59 for total sugar. Both clearance ratio and T/P were reduced to approximately 1 by injection of galactose (2.5 mmol/kg) simultaneously with 2-d-Gal (25 mumol/kg). Phlorizin (2.5 mumol/kg) increased net 2-d-Gal secretion, whereas glucose (2.5 mmol/kg) produced no change in secretion. Both compounds depressed 2-d-Gal T/P. This result suggests the presence of readsorptive transport at the brush border, sensitive to glucose and phlorizin.

Anaerobiosis↗
PubMed1977

The phlorizin effect on the transport of sugars at the antiluminal face of teased flounder tubules.

The effect of phlorizin on the uptake of sugars by teased renal tubules of the flounder (Pseudopleuronectes americanus) and slices of rabbit kidney cortex was investigated: 1. Increasing concentrations (0.05-0.5 mM) of phlorizin inhibited the uptake of D-glucose at the antiluminal face of the tubular cells by affecting primarily the cellular levels of glucose phosphates, whereas the levels of free glucose remained constant. This result suggests the possibility that the actual transport step is associated with sugar phosphorylation. 2. As opposed to the high affinity of phlorizin for the Na+-dependent active transport system for methyl-alpha-D-glucoside in flounder and rabbit kidney, the carrier shared by glucose, 2-deoxyglucose and mannose at the antiluminal face of tubular cells displays a low affinity for the inhibitor. It is suggested that glucose and its C2-analogs enter the renal tubular cells at the antiluminal face by a phosphorylating pathway which has a low affinity for phlorizin, and mix readily with the metabolic pool. This transport pathway serves the nutritional requirements of the cells.

Animals↗
PubMed1977

Renal sugar transport in the winter flounder. III. Two glucose transport systems.

Teased renal tubules of the winter flounder (Pseudopleuronectes americanus) were employed to investigate the structural requirements for two pathways of D-glucose transport which take place preponderantly across the basal (antiluminal) face of renal cells. 1) An inhibition analysis of the equilibrating, Na-independent and phlorizin-sensitive transport of the nonmetabolizable methyl-alpha-D-glucoside (0.1 and 0.5 mM), with 20 glucose analogs (5 mM), was employed to establish the structural requirements for the substrate-carrier interaction: a (pyranose) ring, oxygen, or F at C1, C2-OH, C3-OH, and C4-OH (all axial, 1C model). Some interaction may also occur at C6-OH. D-Glucose shares this transport system. Hydrogen bonding between the oxygens and the carrier is suggested. 2) The phloretin- and phlorizin-sensitive, ouabain-insensitive transport of D-glucose, 2-deoxy-D-glucose, and D-mannose is associated with considerable phosphorylation. The three sugars mutually compete for a shared transport site. The specificity pattern characterizing the transport system defines the following structural requirements: a (pyranose) ring, a free C1-OH, C3-OH, and C4-OH (both axial) and possibly C6-OH. Hydrogen bonding between the carrier and the oxygens at C3, C4, and C6, and covalent bonding at C1 is suggested.

Animals↗