Abstract
Controversial hypotheses exist as to whether hypoxic kidneys produce biologically active erythropoietin (Ep) or an inactive erythropoietic factor that generates Ep from plasma protein in the blood. To clarify the role of the kidney in Ep production we attempted to extract Ep from kidneys of normal and of hypoxia exposed (6 h at 0.42 atm) Sprague-Dawley rats. Ep was measured in the microsomal fraction of kidney homogenates, using the exhypoxic polycythemic mouse assay for Ep. The Ep content was also determined in kidneys that were flushed free of blood with isotonic phosphate-buffer prior to extirpation.
We found 0.04 U Ep/g in blood-depleted kidneys of normal rats. Upon exposure of the animals to hypoxia the Ep level increased to 0.92 U/g kidney. Ep levels were significantly higher in the kidney cortex than in the medulla. The erythropoietic activity in renal extracts was not enhanced after incubation of samples with homologous serum. Ep extracted from hypoxic kidneys behaved identically with plasma-Ep in the following biochemical tests: heat stability, affinity chromatography with wheat germ lectin, ion exchange chromatography, molecular sieve chromatography and neuraminidase inactivation.
These studies support the hypothesis that kidney cortex cells are capable of producing biologically active Ep.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adamson JW (1968) The erythropoietin/hematocrit relationship in normal and polycythemic man: Implications of marrow regulation. Blood 32:597–609
Bauer C, Jelkmann W (1981) Demonstration of high activities of erythropoietin in hypoxic rat kidneys. Pflügers Arch 389:R43
Burlington H, Cronkite EP, Reincke U, Zanjani ED (1972) Erythropoietin production in cultures of goat renal glomeruli. Proc Natl Acad Sci USA 12:3547–3550
Busuttil RW, Roh BL, Fisher JW (1972) Localization of erythropoietin in the glomerulus of the hypoxic dog kidney using a fluorescent antibody technique. Acta Haematol 47:238–242
Contrera JF, Camiscoli JF, Weintraub AH, Gordon AS (1965) Extraction of erythropoietin from kidneys of hypoxic and phenylhydrazine-treated rats. Blood 25:809–816
Cotes PM, Bangham DR (1961) Bioassay of erythropoietin in mice made polycythemic by exposure to air at reduced pressure. Nature 191:1065–1067
Curthoys NP, Bellemann P (1979) Renal cortical cells in primary monolayer culture. Exp Cell Res 121:31–45
Dunn CDR, Jarvis JH, Napier JAF (1976) Changes in erythropoiesis and renal ultrastructure during exposure of mice to hypoxia. Exp Hematol 4:365–381
Erslev AJ (1974) In vitro production of erythropoietin by kidneys perfused with a serum-free solution. Blood 44:77–85
Erslev AJ, Kazal LA (1969) Renal erythropoietic factor. Lack of effect on hypertransfused mice. Blood 34:222–229
Erslev AJ, Caro J, Miller O, Silver R (1980) Plasma erythropoietin in health and disease. Ann Clin Lab Sci 10:250–257
Fisher JW, Birdwell BJ (1961) The production of an erythropoietic factor by the in situ perfused kidney. Acta Haematol 26:224–232
Fisher JW, Taylor G, Porteous DD (1965) Localization of erythropoietin in glomeruli of sheep kidney by fluorescent antibody technique. Nature 205:611–612
Fried W, Kilbridge T, Krantz S, McDonald TP, Lange RD (1969) Studies on extrarenal erythropoietin. J Lab Clin Med 73:244–248
Fried W, Barone-Varelas J, Berman M (1981) Detection of high erythropoietin titers in renal extracts of hypoxic rats. J Lab Clin Med 97:82–86
George WJ, Briggs DW, Rodgers GM, Fisher JW (1977) Metabolism of erythropoietin. In: Kidney Hormones. Vol. II. Erythropoietin (Fisher JW ed.), pp 73–109 New York: Academic Press
Goldwasser E, White WF, Taylor KB (1962) Further purification of sheep plasma erythropoietin. Biochim Biophys Acta 64:487–496
Goldwasser E, Eliason JF, Sikkema D (1975) An assay for erythropoietin in vitro at the milliunit level. Endocrinology 97:315–323
Gordon AS, Kaplan SM (1977) Erythrogenin (REF). In: Fisher JW (ed) Kidney Hormones. Vol II Erythropoietin Academic Press, New York pp 187–229
Gordon AS, Cooper GW, Zanjani ED (1967) The kidney and erythropoiesis. Semin Hematol 4:337–358
Hartcroft PM, Bischoff MB, Bucci TJ (1969) Effects of chronic exposure to high altitude on the juxtaglomerular complex and adrenal cortex of dogs, rabbits and rats. Fed Proc 28:1234–1236
Heim LR (1980) Erythropoietin: What's in a name. Exp Hematol 8:229–230
Hirashima K, Takaku F (1962) Experimental studies on erythropoietin. II. The relationship between juxtaglomerular cells and erythropoietin. Blood 20:1–8
Iscove NN, Sieber F, Winterhalter KH (1974) Erythroid colony formation in cultures of mouse and human bone marrow: Analysis of the requirement for erythropoietin by gel filtration and affinity chromatography on agarose-concanavalin A. J Cell Physiol 83:309–320
Iscove NN, Guilbert LJ, Weyman C (1980) Complete replacement of serum in primary cultures of erythropoietin-dependent red cell precursors (CFU-E) by albumin, transferrin, iron, unsaturated fatty acid, lecithin and cholesterol. Exp Cell Res 126:121–126
Jacobson LO, Goldwasser E, Fried W, Plzak L (1957) Role of the kidney in erythropoiesis. Nature 179:633–634
Kuratowska Z, Lewartowski B, Michalak E (1961) Studies on the production of erythropoietin by isolated perfused organs. Blood 18:527–534
Libbin RM, Person P, Gordon AS (1974) Renal lysosomes: Role in biogenesis of erythropoietin. Science 185:1174–1176
McDonald TP, Martin DH, Simmons ML, Lange RD (1969) Preliminary results of erythropoietin production by bovine kidney cells in culture. Life Sci 8:949–954
Muirhead EE, Leach BE, Fisher JW, Kosinski M (1968) Renal transplantation and extracts and erythropoiesis. Ann NY Acad Sci 149:135–142
Ogle JW, Lange RD, Dunn CDR (1978) Erythropoietin production by rabbit kidney cultures from “programmed” rabbits. Blood 52:233–239
Palade G (1975) Intracellular aspects of the process synthesis. Science 189:347–358
Peschle C, Condorelli M (1975) Biogenesis of erythropoietin: Evidence for pro-erythropoietin in a subcellular fraction of kidney. Science 190:910–912
Putnam FW (1975) Alpha, beta, gamma, omega — the roster of the plasma proteins. In: Putnam FW (ed), The plasma proteins. Vol I. Academic Press, London pp 57–131
Rich IN, Kubanek B (1980) The ontogeny of erythropoiesis in the mouse detected by the erythroid colony-forming technique. J Embryol Exp Morphol 58:143–155
Schooley JC, Mahlmann LJ (1972) Evidence for the de novo synthesis of erythropoietin in hypoxic rats. Blood 40:662–670
Sherwood JB, Goldwasser E (1978) Extraction of erythropoietin from normal kidneys. Endocrinology 103:866–870
Sherwood JB, Robinson SH, Bassan LR, Rosen S, Gordon AS (1972) Production of erythrogenin by organ cultures of rat kidney. Blood 40:189–197
Spivak JL, Small D, Shaper JH, Hollenberg MD (1978) Use of immobilized lectins and other ligands for the partial purification of erythropoietin. Blood 52:1178–1188
Zanjani ED, Contrera JF, Cooper GW, Gordon AS, Wong KK (1967) Renal erythropoietic factor: Role of ions and vasoactive agents in erythropoietin formation. Science 156:1367–1368
Zanjani ED, Gidari AS, Gordon AS, Peschle C (1972) Further studies on the mechanism of erythropoietin formation by the kidney. In: Gordon AS, Condorelli M, Peschle C (eds), Regulation of erythropoiesis. Publishing House Ponte, Milano pp 205–215
Author information
Authors and Affiliations
Additional information
Supported by grants from the Deutsche Forschungsgemeinschaft (Je 95/3 and SFB 43).
Some of the material reported in this paper was presented in preliminary form at the 54. Meeting of the German Physiological Society (Bauer and Jelkmann 1981).
Rights and permissions
About this article
Cite this article
Jelkmann, W., Bauer, C. Demonstration of high levels of erythropoietin in rat kidneys following hypoxic hypoxia. Pflugers Arch. 392, 34–39 (1981). https://doi.org/10.1007/BF00584579
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00584579