Skip to main content
SpringerLink
Log in

Immunolocalization of ecto-5′-nucleotidase in the kidney by a monoclonal antibody

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

A monoclonal antibody IgG, has been raised against ecto-5′-nucleotidase purified from rat kidney homogenate. The specificity of the antibody was verified by immunoprecipitation. The distribution of the corresponding antigen in the rat kidney was studied by immunocytochemistry (FITC and PAP technique) in 1 μm thick cryostat sections. The antibody reacted with the brush border of proximal tubules, the apical cell membrane and the apical cytoplasm of intercalated cells in connecting tubules and collecting ducts and with interstitial cells of the cortex. Among the interstitial cells exclusively stellate shaped fibroblasts were reactive whereas rounded interstitial cells (type II interstitial cells) as well as pericytes and endothelial cells of peritubular capillaries were unreactive. Compared to the staining intensity of the fibroblasts in the cortical labyrinth the reactivity of the fibroblasts in the medullary rays of the cortex was weak or absent. Interstitial cells of the entire medulla were unreactive. Concerning the fibroblasts in the periarterial connective tissue, those surrounding the larger arteries (arcuate arteries, cortical radial arteries) were negative, those alongside afferent and efferent arterioles were positive. Endothelia of lymphatic capillaries travelling within the periarterial connective tissue were also positive. All components of the juxtaglomerular apparatus were negative.

The findings are consistent with an interstitial production of adenosine, available extracellularly and thus being able to reach the major target sites of adenosine, the smooth muscles of glomerular arterioles, including the granular cells at the glomerular vascular pole.

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

  • Albertine KH, O'Morchoe CCC (1979) Distribution and density of the canine renal cortical lymphatic system. Kidney Int 16:470–480

    Google Scholar 

  • Arch JRS, Newsholme EA (1979) The control of the metabolism and the hormonal role of adenosine. Essays Biochem 14:82–123

    Google Scholar 

  • Arend LJ, Sonnenburg WK, Smith WL, Spielman WS (1987) A1 and A2 adenosine receptors in rabbit cortical collecting tubule cells. J Clin Invest 79:710–713

    Google Scholar 

  • Berne RM, Rall TW, Ribio R (1983) Regulatory function of adenosine. Martinus Nijhoff, Boston, Mass., USA

    Google Scholar 

  • Bohman SO (1983) The ultrastructure of the renal interstitium. In: Brenner BM, Stein JH (eds) Contemporary issues in nephrology. Churchill Livingstone, New York, pp 1–34

    Google Scholar 

  • Bohman SO, Sundelin B, Forsum U, Tribukait B (1988) Experimental depletion of different renal interstitial cell. Am J Med Sci 295:252–257

    Google Scholar 

  • Churchill PC, Bidani AK (1982) Hypothesis: adenosine mediates hemodynamic changes in renal failure. Med Hypotheses 8:275–285

    Google Scholar 

  • Cronstein BN, Levin RI, Belanoff J, Weissmann G, Hirschhorn R (1986) Adenosine: an endogenous inhibitor of neutrophil-mediated injury to endothelial cells. J Clin Invest 78:760–770

    Google Scholar 

  • Dawson TP, Gandhi R, Le Hir M, Kaissling B (1989) Ecto 5′-Nucleotidase: localization by light microscopic histochemistry and immuno-histochemistry methods in the rat kidney. J Histochem Cytochem 37:39–47

    Google Scholar 

  • Dobson JG (1983) Mechanism of adenosine inhibition of catechol-amine-induced responses in heart. Circ Res 52:151–160

    Google Scholar 

  • Engler R (1987) Consequences of activation and adenosine-mediated inhibition of granulocytes during myocardial ischemia. Fed Proc 46:2407–2412

    Google Scholar 

  • Fazekas de St Groth, Scheidegger D (1980) Production of monoclonal antibodies: strategy and tactics. J Immunol Methods 35:1–21

    Google Scholar 

  • Franco MP, Bell PD, Navar LG (1989) Effect of adenosine A, analogue on tubulo-glomerular feedback mechanism. Am J Physiol 257:F231-F236

    Google Scholar 

  • Galfre G, Howe SC, Milstein C (1977) Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature 266:550–552

    Google Scholar 

  • Gürner AC, Smith J, Cattel V (1987) The origin of Ia antigen-expressing cells in the rat kidney. Am J Pathol 127:342–348

    Google Scholar 

  • Hall JE, Granger JP, Hester RL (1985) Interactions between adenosine and angiotensin II in controlling glomerular filtration. Am J Physiol 248:F340-F346

    Google Scholar 

  • Hall JE, Granger JP (1986) Renal hemodynamics and arterial pressure during chronic intrarenal adenosine infusion in conscious dogs. Am J Physiol 250:F32-F39

    Google Scholar 

  • Hart DNJ, Fabre JW (1981a) Demonstration and characterization of Ia-positive dentritic cells in the interstitial connective tissues of rat heart and other tissues, but not brain. J Exp Med 153:347–361

    Google Scholar 

  • Hart DNJ, Fabre JW (1981b) Major histocompatibility complex antigens in rat kidney, ureter, and bladder. Transplantation 31:318–325

    Google Scholar 

  • Hauri HP, Sterchi EE, Bienz D, Fransen JAM, Marxer A (1985) Expression and intracellular transport of microvillus membrane hydrolases in human intestinal epithelial cells. J Cell Biol 101:838–851

    Google Scholar 

  • Headrick JP, Willis RJ (1989) 5′-Nucleotidas activity and adenosine formation in stimulated, hypoxic and unperfused rat heart. Biochem J 261:541–550

    Google Scholar 

  • Holz FG, Steinhausen M (1987) Renovascular effects of adenosine receptor agonists. Renal Physiol 10:272–282

    Google Scholar 

  • Joyner WL, Mohama RE, Myers TO, Gilmore JP (1988) The selective response to adenosine of renal microvessels from hamster explants. Microvasc Res 35:122–131

    Google Scholar 

  • Kaissling B, Kriz W (1982) Variability of intercellular spaces between macula densa cells: TEM study in rabbits and rats. Kidney Int Suppl 22:S9-S17

    Google Scholar 

  • Koury ST, Bondurant MC, Koury MJ (1988) Localization of erythropoietin synthesizing cells in murine kidneys by in situ hybridization. Blood 71:524–527

    Google Scholar 

  • Koury ST, Koury MJ, Bondurant MC, Caro J, Graber SE (1989) Quantitation of erythropoietin-producing cells in kidneys of mice by in situ hybridization: correlation with hematocrit, renal erythropoietin mRNA, and serum erythropoietin concentration. Blood 74:645–651

    Google Scholar 

  • Kriz W, Dieterich HJ (1970) Das Lymphgefäßsystem der Niere bei einigen Säugetieren. Licht- und elektronenmikroskopische Untersuchungen. Z Anat Entwicklungsgesch 131:111–147

    Google Scholar 

  • Kriz W (1987) A periarterial pathway for intrarenal distribution of renin. Kidney Int 31:S51-S56

    Google Scholar 

  • Kurtz A, Bruna R della, Pfeilschifter J, Bauer CH (1988) Role of cGMP as second messenger of adenosine in the inhibition of renin release. Kidney Int 33:798–803

    Google Scholar 

  • Lacombe C, Silva JL da, Bruneval P, Fournier JG, Wendling F, Casadevall N, Camilleri JP, Bariety J, Varet B, Tambourin P (1988) Peritubular cells are the site of erythropoietin synthesis in the murine hypoxic kidney. J Clin Invest 81:620–623

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Le Hir M, Kaissling B (1989) Distribution of 5′-nucleotidase in the renal interstitium of the rat. Cell Tissue Res 258:177–182

    Google Scholar 

  • Le Hir M, Gandhi R, Dubach UC (1989) Purification and properties of a 5′-nucleotidase from rat renal membranes. Enzyme 41:87–93

    Google Scholar 

  • Le Hir M, Kaissling B, Gandhi R, Dubach UC (1989) Fibroblasts may represent the main site of production of interstitial adenosine in the kidney. Kidney Int 36:319–320

    Google Scholar 

  • Lietzke R, Unsicker K (1985) A statistical approach to determine monoclonality after limiting cell plating of a hybridoma clone. J Immunol Methods 76:223–228

    Google Scholar 

  • Miller WL, Thomas RA, Berne RM, Rubio R (1978) Adenosine production in the ischemic kidney. Circ Res 43:390

    Google Scholar 

  • Mills DCB, McFarlane DE, Lemmex BWG, Haslam RJ (1983) Receptors for nucleosides and nucleotides on blood platelets. In: Berne RM, Rall TW, Rubio R (eds) Regulatory function of adenosine. Martinus and Nijhoff, Boston, pp 277–289

    Google Scholar 

  • Naito Y, Lowenstein JM (1981) 5′-Nucelotidase of rat heart. Biochemistry 20:5194

    Google Scholar 

  • Newby AC (1988) The pigeon heart 5′-nucleotidase responsible for ischemia-induced adenosine formation. Biochem J 253:123–130

    Google Scholar 

  • Osswald H, Hermes HH, Nabakowski G (1982) Role of adenosine in signal transmission of tubulo-glomerular feedback. Kidney Int 22 [Suppl 12]:S136-S142

    Google Scholar 

  • Osswald H, Nabakowski G, Hermes H (1980) Adenosine as a possible mediator of metabolic control of glomerular filtration rate. Int J Biochem 12:263–267

    Google Scholar 

  • Pawlowska D, Granger JP, Knox FG (1987) Effects of adenosine infusion into renal interstitium on renal hemodynamics. Am J Physiol 252:F678-F682

    Google Scholar 

  • Paul Ph, Rothmann SA, Meagher RC (1988) Modulation of erythropoietin production by adenosine. J Lab Clin Med 112:168–173

    Google Scholar 

  • Ramos-Salazar A, Baines AD (1986) Role of 5′-nucleotidase in adenosine-mediated renal vasoconstriction during hypoxia. J Pharmacol Exp Ther 236:494–499

    Google Scholar 

  • Schnermann J (1988) Effect of adenosine analogues on tubuloglomerular feedback responses. Am J Physiol 255:F33–42

    Google Scholar 

  • Schnermann J, Davis JA, Briggs JP (1990a) Adenosine-angiotensin interactions in the control of afferent arteriolar resistance. Kidney Int 37:556

    Google Scholar 

  • Schnermann J, Weihprecht H, Briggs JP (1990b) Inhibition of tubulo-glomerular feedback during adenosine receptors blockade. Am J Physiol 285:F553-F561

    Google Scholar 

  • Schurek HJ (1988) Die Nierenmarkhypoxie: Ein Schlüssel zum Verständnis des akuten Nierenversagens? Klin Wochenschr 66:828–835

    Google Scholar 

  • Skott O, Baumbach L (1985) Effects of adenosine on renin release from isolated rat glomeruli and kidney slices. Pflügers Arch 404:232–237

    Google Scholar 

  • Spielman WS, Osswald H (1979) Blockade of postocclusive renal vasoconstriction by an angiotensin II antagonist: evidence for an angiotensin-adenosine interaction. Am J Physiol 237:F463-F467

    Google Scholar 

  • Spielman WS, Thompson CI (1982) A proposed role for adenosine in the regulation of renal hemodynamics and renin release. Am J Physiol 242:F423–435

    Google Scholar 

  • Steiniger B, Klempnauer J, Wonigeit K (1984) Phenotype and histological distribution of interstitial dendritic cells in the rat pancreas, liver, heart, and kidney. Transplantation 38:169–175

    Google Scholar 

  • Stewart JR, Kenny AJ (1984) Proteins of the kidney microvillar membrane. Biosynthesis of endopeptidase-24.11, dipeptidylpeptidase IV and aminopeptidases N and A in pig kidney slices. Biochem J 224:549–558

    Google Scholar 

  • Swann HG, Norman HJ (1970) The periarterial spaces of the kidney. Tex Rep Biol Med 28:317–334

    Google Scholar 

  • Ueno M, Brookins J, Beckman B, Fisher JW (1988) A1 and A2 adenosine receptor regulation of erythropoietin production. Life Sci 43:229–237

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department Forschung and Medizinische Poliklinik, Kantonsspital, CH-4031, Basel, Switzerland

    R. Gandhi & M. Le Hir

  2. Anatomisches Institut der Universität, Winterthurerstrasse 190, CH-8057, Zürich-Irchel, Switzerland

    B. Kaissling

Authors
  1. R. Gandhi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Le Hir
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B. Kaissling
    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

Gandhi, R., Le Hir, M. & Kaissling, B. Immunolocalization of ecto-5′-nucleotidase in the kidney by a monoclonal antibody. Histochemistry 95, 165–174 (1990). https://doi.org/10.1007/BF00266589

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00266589

Keywords

Search

Navigation