Skip to main content
SpringerLink
Log in

Substrate utilization in the isolated perfused cortical thick ascending limb of rabbit nephron

  • Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Isolated segments of cortical thick ascending limbs (cTAL) of rabbit kidney were perfused in vitro and the equivalent short circuit current (Isc) was measured. In a first series all substrates were removed on either side. Isc fell rapidly to 50±12% after 3 min and to 27±6% (n=5) after 10 min. This indicates that in cTAL segments Isc is strictly dependent on the presence of substrates. In series two it was tested what substrates can be utilized by the cTAL segment, and from which epithelial side [bath (b) or lumen (l)] the substrates are taken up. From the l-side only butyrate (10 mmol · l−1) sustained the Isc at 95±2% (n=7). All other tested substrates (10 mmol · l−1): pyruvate, acetate, β-OH-butyrate,d-glucose, andl-lactate lead to a marked decline in Isc. From the b-side several substrates (5–10 mmol · l−1) sustained the Isc:d-glucose,d-mannose, butyrate, β-OH-butyrate, acetoacetate,l-lactate, acetate and pyruvate. Other compounds (1–10 mmol · l−1): citrate, α-ketoglutarate, succinate, glutamate, glutamine, propionate, caprylate and oleate did not sustain Isc. In the third series the mechanism of substrate utilization from the basolateral cell side was studied. It was shown that the Isc is a saturable function of thed-glucose,l-lactate, acetate, pyruvate or β-OH-butyrate concentration with apparentK m's between 0.05–1.0 mmol · l−1. Several known inhibitors of sugar and of anion transport were tested at the bath side: phlorrhizin was without effect. Phloretin (500 μmol · l−1) inhibited Isc by 96%, yet its effect was not dependent on the presence of substrates on the b-side since inhibition ocurred also if the b-perfusate contained no substrate and Isc was driven by luminal butyrate. Also SITS (5 mmol · l−1) exerted only a small inhibitory effect which was not specific since it was also observed with luminal butyrate. α-Cyano-m-OH-cinnamate (10 mmol · l−1) inhibited the Isc specifically whenl-lactate was the bath substrate. Probenecid (1 mmol · l−1) had a similar yet less marked inhibitory effect. Thed-glucose uptake from the b-side was specifically inhibited by cytochalasin B at 5 · 10−6 mol · l−1. We conclude that the cTAL segment of the rabbit utilizesd-glucose and/or small anions such as pyruvate orl-lactate or acetate to energize salt reabsorption. The link between substrate availability and salt reabsorption is extremely close in this nephron segment. Substrate uptake occurs from the blood side. Sugar uptake can be inhibited by cytochalasin B andl-lactate uptake by probenecid and α-cyano-m-OH-cinnamat. These data suggest that substrate uptake at the basolateral cell side occurs probably via carrier systems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Andersen OS, Finkelstein A, Katz I, Cass A (1976) Effect of phloretin on the permeability in thin lipid membranes. J Gen Physiol 67:749–771

    Google Scholar 

  2. Baeyer H v (1981) Transport ofd-glucose in the mammalian kidney. In: Greger R, Lang F, Silbernagl S (eds) Renal transport of organic substances. Springer, Berlin Heidelberg New York, pp 154–177

    Google Scholar 

  3. Barac-Nieto M, Murer H, Kinne R (1982) Asymmetry in the transport of lactate by basolateral and brush border membranes of rat kidney cortex. Pflügers Arch 392:366–371

    Google Scholar 

  4. Burg MB, Grantham J, Abramow M, Orloff J (1966) Preparation and study of fragments of single rabbit nephrons. Am J Physiol 210 (6):1293–1298

    Google Scholar 

  5. Chance B (1980) Non-invasive approaches to the biomedical basis for physiological function. Proc Int Union Physiol Sci 14:14–15

    Google Scholar 

  6. Cohen JJ, Barac-Nieto M (1973) Renal metabolism of substrates in relation to renal function. In: Orloff J, Berliner RW, Geiger SR (eds) Handbook of physiology: Renal physiology. Am Physiol Soc, Washington DC, pp 909–1001

    Google Scholar 

  7. Cohen JJ, Kamm DE (1976) Renal metabolism: Relation to renal function. In: Brenner BM, Rector FC Jr (eds) The kidney. Saunders Comp, Philadelphia London Toronto, pp 126–214

    Google Scholar 

  8. De Jonge PC, Wieringa T, Putten JPM Van, Krans HMJ, Dam K Van (1983) Phloretin — an uncoupler and an inhibitor of mitochondrial oxidative phosphorylation. Biochim Biophys Acta 722:219–225

    Google Scholar 

  9. Eveloff J, Bayerdörffer, Silva P, Kinne R (1981) Sodium-chloride transport in the thick ascending limb of Henle's loop. Oxygen consumption studies in isolated cells. Pflügers Arch 389:263–270

    Google Scholar 

  10. Forman SA, Verkman AS, Dix JA, Solomon AK (1982) Interaction of phloretin with the anion transport protein of the red blood cell membrane. Biochim Biophys Acta 689:531–538

    Google Scholar 

  11. Frömter E (1982) Electrophysiological analysis of rat renal sugar and amino acid transport. I. Basic phenomena. Pflügers Arch 393:179–189

    Google Scholar 

  12. Greger R (1981a) Cation selectivity of the isolated perfused cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 390:30–37

    Google Scholar 

  13. Greger R (1981b) Coupled transport of Na+ and Cl− in the thick ascending limb of Henle's loop of rabbit kidney. In: Klinke R, Lahn W, Querfurth J, Scholtholt J (eds) Scand Audiol Suppl 14:1–15

  14. Greger R, Hampel W (1981) A modified system for in vitro perfusion of isoated renal tubules. Pflügers Arch 389:175–176

    Google Scholar 

  15. Greger R, Schlatter E (1983a) Properties of the basolateral membrane of the cortical thick ascending limb of Henle's loop of rabbit kidney. A model for secondary active chloride transport. Pflügers Arch 396:325–334

    Google Scholar 

  16. Greger R, Schlatter E (1983b) Cellular mechanism of the action of loop diuretics on the thick ascending limb of Henle's loop. Klin Wochenschr 61:1019–1027

    Google Scholar 

  17. Greger R, Schlatter E (1983c) Properties of the lumen membrane of the cortical thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 396:315–324

    Google Scholar 

  18. Griffin JF, Rampal AL, Jung CH (1982) Inhibition of glucose transport in human erythrocytes by cytochalasins: A model based on diffraction studies. Proc Natl Acad Sci USA 79: 3759–3763

    Google Scholar 

  19. Guder WG, Schmidt U (1976) Substrate and oxygen dependence of renal metabolism. Kidney Int 10:32–38

    Google Scholar 

  20. Guder WG, Wirthensohn G (1981) Renal turnover of substrates. In: Greger R, Lang F, Silbernagl S (eds) Renal transport of organic substances. Springer, Berlin Heidelberg New York, pp 65–77

    Google Scholar 

  21. Guder WG, Ross BD (1984) Enzyme distribution along the nephron. Kidney Int (in press)

  22. Häberle DA (1981) Characteristics of p-aminohippurate transport in the mammalian kidney. In: Greger R, Lang F, Silbernagl S (eds) Renal transport of organic substances. Springer, Berlin Heidelberg New York, pp 189–209

    Google Scholar 

  23. Hebert SC, Culpepper RM, Andreoli TE (1981) NaCl transport in mouse medullary thick ascending limbs. I. Functional nephron heterogeneity and ADH-stimulated NaCl cotransport. Am J Physiol 241:F412-F431

    Google Scholar 

  24. Hopfer U, Sigrist-Nelson K, Amman E, Murer H (1976) Differences in neutral amino acid and glucose transport between brush border and basolateral plasma membrane of intestinal epithelial cells. J Cell Physiol 89:805–810

    Google Scholar 

  25. Höhmann B, Zwiebel R, Yamagata A, Kinne R (1969) Enzymaktivitäten im isolierten proximalen Tubulus der Kaninchenniere. Pflügers Arch 312:110–125

    Google Scholar 

  26. Kalckar H (1936) Inhibitory effect of phloridzin and phloretin on kidney phosphatase. Nature 138:289

    Google Scholar 

  27. Kimmich GA, Randles J (1975) A Na+-independent, phloretin-sensitive monosaccharide transport system in isolated intestinal epithelial cells. J Membr Biol 23:57–76

    Google Scholar 

  28. Kinne R, Murer H (1976) Polarity of epithelial cells in relation to transepithelial transport in kidney and intestine. In: Robinson JWL (ed) Intestinal ion transport. MTP Press Ltd., Lancaster, pp 79–95

    Google Scholar 

  29. Kinne R, Murer H, Kinne-Saffran E, Thees M, Sachs G (1975) Sugar transport by renal plasma membrane vesicles. Characterization of the systems in the Brush-Border Microvilli and basal-lateral plasma membranes. J Membr Biol 21:375–395

    Google Scholar 

  30. Klein KL, Wang M-S, Torikai S, Davidson WD, Kurokawa K (1981) Substrate oxidation by isolated single nephron segments of the rat. Kidney Int 20:29–35

    Google Scholar 

  31. Kurokawa K, Torikai S, Wang M-S, Klein KL (1982) Metabolic heterogeneity of the nephron. Mineral Electrolyte Metab 77:225–236

    Google Scholar 

  32. Le Bouffant F, Hus-Citharel A, Morel F (1982) In vitro14CO2 production by single pieces of rat cortical thick ascending limbs and its coupling to active salt transport. In: Morel F (ed) Biochemistry of kidney functions, INSERM Symposium 21. Elsevier Biomedical Press, Amsterdam New York Oxford, pp 363–370

    Google Scholar 

  33. LeFevre PG, Marshall JK (1959) The attachment of phloretin and analogues to human erythrocytes in connection with inhibition of sugar transport. J Biol Chem 234 (11):3022–3026

    Google Scholar 

  34. Le Hir M, Dubach UC (1982) Activities of enzymes of the tricarboxylic acid cycle in segments of the rat nephron. Pflügers Arch 395:239–243

    Google Scholar 

  35. Macey RI, Farmer REL (1970) Inhibition of water and solute permeability in human red cells. Biochim Biophys Acta 211:104–106

    Google Scholar 

  36. Murer H, Barac-Nieto M, Ullrich KJ, Kinne R (1981) Renal transport of lactate. In: Greger R, Lang F, Silbernagl S (eds) Renal transport of organic substances. Springer, Berlin Heidelberg New York, pp 210–223

    Google Scholar 

  37. Murer H, Kinne R (1977) Sidedness and coupling of transport in small intestinal and renal epithelia. In: Semenza G, Carafoli E (eds) Biochemistry of membrane transport, FEBS-Symposium No. 42. Springer, Berlin Heidelberg New York, pp 292–304

    Google Scholar 

  38. Owen JD (1974) The effect of phloretin on the potassium conductance in Aplysia giant neurons. J Membr Biol 16:65–78

    Google Scholar 

  39. Owen JD, Solomon AK (1972) Control of nonelectrolyte permeability in red cells. Biochim Biophys Acta 290:414–418

    Google Scholar 

  40. Owen A, Caplan SR, Essig A (1975) A comparison of the effects of ouabain and 2-deoxy-d-glucose on the thermodynamic variables of the frog skin. Biochim Biophys Acta 394:438–448

    Google Scholar 

  41. Petersen K-U, Wood JR, Schulze G, Heintze K (1981) Stimulation of gallbladder fluid and electrolyte absorption by butyrate. J Membr Biol 62:183–193

    Google Scholar 

  42. Randles J, Kimmich GA (1978) Effects of phloretin and theophylline on 3-O-methyl-glucose transport by intestinal epithelial cells. Am J Physiol 234 (3):C64-C72

    Google Scholar 

  43. Reyes J, Greco F, Otais R, Latorre R (1983) Phloretin and phloretin analogs: Mode of action in planar lipid bilayers and monolayers. J Membr Biol 72:93–103

    Google Scholar 

  44. Schlatter E, Greger R (1982) Metabolic substrates for maintaining active NaCl transport in the isolated cortical thick ascending limb (cTAL) of rabbit kidney. Pflügers Arch 394:R22

    Google Scholar 

  45. Schmidt I, Guder WG (1976) Sites of enzyme activity along the nephron. Kidney Int 9:233–242

    Google Scholar 

  46. Shimada H, Endou H, Sakai F (1982) Distribution of α-glutamyl transpeptidase and glutaminase isoenzymes in the rabbit single nephron. Jpn J Pharmacol 32:121–129

    Google Scholar 

  47. Silverman M, Turner RJ (1982) 2-Deoxy-d-glucose transport in dog kidney. Am J Physiol 242:F711-F720

    Google Scholar 

  48. Ullrich KJ (1976) Renal tubular mechanisms of organic solute transport. Kidney Int 9:134–148

    Google Scholar 

  49. Ullrich KJ (1979) Sugar, amino acid, and Na+ cotransport in the proximal tubule. Ann Rev Physiol 41:181–195

    Google Scholar 

  50. Ullrich KJ, Fasold H, Rumrich G, Klöss S (1984) Secretion and contraluminal uptake of dicarboxylic acids in the proximal convolution of rat kidney. Pflügers Arch 400:241–249

    Google Scholar 

  51. Ullrich KJ, Radtke HW, Rumrich G (1971) The role of bicarbonate and other buffers on isotonic fluid absorption in the proximal convolution of the rat kidney. Pflügers Arch 330:149–161

    Google Scholar 

  52. Ullrich KJ, Rumrich G, Klöss S (1982) Reabsorption of monocarboxylic acids in the proximal tubule of the rat kidney. I. Transport kinetics ofd-lactate, Na+-dependence, pH-dependence and effect of inhibitors. Pflügers Arch 395:212–219

    Google Scholar 

  53. Ullrich KJ, Rumrich G, Klöss S, Fasold H (1982) Reabsorption of monocarboxylic acid in the proximal tubule of the rat kidney. III. Specificity for aromatic compounds. Pflügers Arch 395:227–231

    Google Scholar 

  54. Vandewalle A, Wirthensohn G, Heidrich H-G, Guder WG (1981) Distribution of hexokinase and phosphoenolpyruvate carboxykinase along the rabbit nephron. Am J Physiol 240:F492-F500

    Google Scholar 

  55. Warnock DG, Greger R, Dunham PB, Frizzell RA, Field M, Spring KR, Ives HE, Aronsen PS, Seifter J (1984) Ion transport processes in apical membranes of epithelia. Fed Proc (in press)

  56. Wright EM (1974) Active transport of iodide and other anions across the choroid plexus. J Physiol 240:535–566

    Google Scholar 

  57. Wright EM, Van Os CH, Mircheff AK (1980) Sugar uptake by intestinal basolateral membrane vesicles. Biochim Biophys Acta 597:112–124

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Max-Planck-Institut für Biophysik, Kennedyallee 70, D-6000, Frankfurt/Main, Germany

    Monika Wittner, Claudia Weidtke, Eberhard Schlatter, Antonio di Stefano & Rainer Greger

Authors
  1. Monika Wittner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Claudia Weidtke
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eberhard Schlatter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Antonio di Stefano
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rainer Greger
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wittner, M., Weidtke, C., Schlatter, E. et al. Substrate utilization in the isolated perfused cortical thick ascending limb of rabbit nephron. Pflugers Arch. 402, 52–62 (1984). https://doi.org/10.1007/BF00584832

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00584832

Key words

Search

Navigation