Advertisement

Pflügers Archiv

, Volume 408, Issue 5, pp 479–483 | Cite as

Effects of glutaraldehyde fixation on renal tubular function

I. Preservation of vasopressin-stimulated water and urea pathways in rat papillary collecting duct
  • Yoshiaki Kondo
  • Masashi Imai
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands

Abstract

Using the in vitro microperfusion technique on isolated rat papillary collecting duct (PCD), we examined whether the glutaraldehyde-fixation method can be also applied to the mammalian collecting duct for preservation of the vasopressin-stimulated water and urea transport. Arginine vasopressin (AVP) at 10−9 mol/l increased diffusional water permeability (Pdw) from 101.9±10.76 to 283.3±16.67×10−7 cm2 s−1 (n=8,P<0.01) and urea permeability (Purea) from 30.3±2.24 to 83.5±7.80×10−7 cm2 s−1 (n=8,P<0.01). Both parameters remained elevated after fixation with 0.1 mol/l glutaraldehyde even in the absence of AVP, with the values being 265.0±14.47 and 74.5±7.15×10−7 cm2 s−1, respectively. Glutaraldehyde fixation did not affect the basal levels ofPdw orPurea. Phloretin at 2.5×10−4 mol/l decreased glutaraldehyde-fixed AVP-stimulatedPurea from 79.0±7.96 to 29.7±3.66×10−7 cm2 s−1 (n=4,P<0.01) and from 73.2±7.05 to 38.7±3.53×10−7 cm2 s−1 (n=4,P<0.01) when the drug was added to the lumen or to the bath, respectively. Phloretin also decreased glutaraldehyde-fixed non-stimulatedPurea by 25–40%. However, this drug did not affect glutaraldehyde-fixedPdw. These findings indicate that the glutaraldehyde fixation method can be applied to mammalian collecting tubules for studying vasopressin stimulatedPdw andPurea.Purea fixed by glutaraldehyde is functionally flexible and may be distinct from the water pathway.

Key words

Papillary collecting duct Vasopressin Phloretin Water channel Urea channel Glutaraldehyde 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Burg MB, Granthan J, Abramow M, Orloff J (1966) Preparation and study of fragments of single rabbit nephron. Am J Physiol 210:1293–1298Google Scholar
  2. 2.
    Carvounis CP, Franki N, Levine SD, Hays RM (1979) Membrane pathways for water and solutes in the toad bladder. I. Independent activation of water and urea transport. J Membr Biol 49:253–268Google Scholar
  3. 3.
    Carvounis CP, Levine SD, Franki N, Hays RM (1979) Membrane pathways for water and solutes in the toad bladder. II. Reflexion coefficients of water and solute channels. J Membr Biol 49:269–287Google Scholar
  4. 4.
    Chevalier J, Bourguet J, Hugon JS (1974) Membrane associated particles: distribution in frog urinary bladder epithelium at rest and after oxitocin treatment. Cell Tissue Res 152: 129–140Google Scholar
  5. 5.
    Curci S, Casavola V, Cremaschi D, Lippe C (1976) Facilitated transport of urea across the toad gall bladder. Pflügers Arch 362:109–112Google Scholar
  6. 6.
    Eggena P (1972) Glutaraldehyde fixation method for determining the permeability to water of the toad urinary bladder. Endocrinology 91:240–246Google Scholar
  7. 7.
    Eggena P (1972) Osmotic regulation of toad bladder responsiveness to neurohypophyseal hormones. J Gen Physiol 60:665–678Google Scholar
  8. 8.
    Eggena P (1973) Inhibition of vasopressin-stimulated urea transport across the toad bladder by thiourea. J Clin Invest 52:2963–2970Google Scholar
  9. 9.
    Eggena P (1983) Effect of glutaraldehyde on hydrosmotic response of toad bladder to vasopressin. Am J Physiol 244:C37-C43Google Scholar
  10. 10.
    Eggena P, Gibas A (1983) Activation energy for water transport in toad bladder. Am J Physiol 244:C44-C49Google Scholar
  11. 11.
    Hall DA, Grantham JJ (1980) Temperature effect on ADH response of isolated perfused rabbit collecting tubules. Am J Physiol 239:F595-F601Google Scholar
  12. 12.
    Hardy M (1985) Urea and Na+ permeability in toad urinary bladder: one or two solute pathways?. Am J Physiol 248:F 56-F 63Google Scholar
  13. 13.
    Hardy M, DiBona DR (1982) Microfilament and hydrosmotic action of vasopressin in toad urinary bladder. Am J Physiol 243:C 200-C 204Google Scholar
  14. 14.
    Hardy M, DiBona DR (1982) Extracellular Ca2+ and the effect of antidiuretic hormone on the water permeability. J Membr Biol 67:27–44Google Scholar
  15. 15.
    Kawamura S, Kokko JP (1976) Urea secretion by the straight segment of the proximal tubules. J Clin Invest 58:604–612Google Scholar
  16. 16.
    Levine S, Franki N, Hays RM (1973) Effect of phloretin on water and solute movement in the toad bladder. J Clin Invest 52:1435–1442Google Scholar
  17. 17.
    Macey RJ, Farmer REL (1970) Inhibition of water and solute permeability in human red cells. Biochim Biophys Acta 211:104–106Google Scholar
  18. 18.
    Morgan T, Berliner RW (1968) Permeability of the loop of Henle, vasa recta, and collecting duct to water, urea and sodium. Am J Physiol 215:108–115Google Scholar
  19. 19.
    Muller J, Kachadorian WA, DiScala VA (1980) Evidence that ADH-stimulated intramembrane particle aggregate are transferred from cytoplasmic to luminal membranes in toad bladder epithelial cells. J Cell Biol 85:83–95Google Scholar
  20. 20.
    Parisi M, Merot J, Bourguet J (1985) Glutaraldehyde fixation preserves the permeability properties of the ADH-induced water channels. J Membr Biol 86:239–245Google Scholar
  21. 21.
    Petrucelli RJ, Eggena P (1982) Improtance of molecular size and hydrogen bonding in vasopressin-stimulated urea transport. Am J Physiol 243:C 27-C 34Google Scholar
  22. 22.
    Rapoport J, Kachadorian WA, Muller J, Franki N, Hays RM (1981) Stabilization of vasopressin-induced membrane events by bifunctional imidoesters. J Cell Biol 89:261–266Google Scholar
  23. 23.
    Rocha AS, Kokko JP (1974) Permeability of medullary nephron segments to urea and water: effect of vasopressin. Kidney Int 6:376–387Google Scholar
  24. 24.
    Rocha AS, Kudo LH (1982) Water, urea, sodium, chloride, and potassium transport in the in vitro isolated perfused papillary collecting duct. Kidney Int 22:485–491Google Scholar
  25. 25.
    Schafer JA, Andreoli TE (1972) The effect of antidiuretic hormone on solute flows in mammalian collecting tubules. J Clin Invest 51:1279–1286Google Scholar
  26. 26.
    Schuchter SH, Franki N, Hays RM (1973) The effect of tanning agents on the permeability of the toad bladder to water and solutes. J Membr Biol 14:177–191Google Scholar
  27. 27.
    Wade JB, Stetson DL, Lewis SA (1981) ADH action: evidence for a membrane shuttle mechanism. Ann NY Acad Sci 372:106–117Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Yoshiaki Kondo
    • 1
  • Masashi Imai
    • 1
  1. 1.Department of PharmacologyNational Cardiovascular Center, Research InstituteSuita, OsakaJapan

Personalised recommendations