Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1985 Oct;76(4):1643–1648. doi: 10.1172/JCI112149

Activity in fetal bovine serum that stimulates erythroid colony formation in fetal mouse livers is insulinlike growth factor I.

A Kurtz, W Härtl, W Jelkmann, J Zapf, C Bauer
PMCID: PMC424153  PMID: 4056043

Abstract

In the present study, the erythropoietic activity of fetal serum was characterized. Using fetal bovine serum (FBS) as a source of the erythropoietic activity and serum-free cultures of fetal mouse livers (FMLC assay) as a detection system, we found that FBS stimulated colony formation from late erythroid progenitor cells (CFU-E) in a dose-dependent fashion. The slope of the dose-response curve, however, was significantly different from that for erythropoietin (Ep), the best-characterized erythropoietic activity so far. The erythropoietic activity of FBS was found in the 120-160- and 40-70-kD range at neutral pH. In the presence of 1 M acetic acid, however, the erythropoietic activity had an apparent molecular mass between 3 and 13 kD. From ion exchange experiments with DEAE-cellulose, the isoionic point of the activity was estimated to about pH 5. Furthermore, the erythropoietic activity of FBS was found to be co-eluted on Sephadex G-150 with the binding proteins of insulinlike growth factors (IGF). The IGF I concentration determined by radioimmunoassay was 70 ng IGF I/ml. The Ep activity of FBS was less than 5 mU/ml when determined with the posthypoxic polycythemic mouse assay for Ep. These results suggest that the erythropoietic activity of FBS is related to IGF and not to Ep. The erythropoietic activity of FBS was abolished by an antiserum against IGF I. Furthermore, IGF I was a factor of approximately 40 more potent than IGF II in stimulating erythroid colony formation. All of these findings suggest that the erythropoietic activity of FBS is IGF I.

Full text

PDF
1643

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Daughaday W. H., Parker K. A., Borowsky S., Trivedi B., Kapadia M. Measurement of somatomedin-related peptides in fetal, neonatal, and maternal rat serum by insulin-like growth factor (IGF) I radioimmunoassay, IGF-II radioreceptor assay (RRA), and multiplication-stimulating activity RRA after acid-ethanol extraction. Endocrinology. 1982 Feb;110(2):575–581. doi: 10.1210/endo-110-2-575. [DOI] [PubMed] [Google Scholar]
  2. Daughaday W. H., Ward A. P., Goldberg A. C., Trivedi B., Kapadia M. Characterization of somatomedin binding in human serum by ultracentrifugation and gel filtration. J Clin Endocrinol Metab. 1982 Nov;55(5):916–921. doi: 10.1210/jcem-55-5-916. [DOI] [PubMed] [Google Scholar]
  3. Erslev A. J., Caro J., Kansu E., Silver R. Renal and extrarenal erythropoietin production in anaemic rats. Br J Haematol. 1980 May;45(1):65–72. doi: 10.1111/j.1365-2141.1980.tb03811.x. [DOI] [PubMed] [Google Scholar]
  4. Froesch E. R., Schmid C., Schwander J., Zapf J. Actions of insulin-like growth factors. Annu Rev Physiol. 1985;47:443–467. doi: 10.1146/annurev.ph.47.030185.002303. [DOI] [PubMed] [Google Scholar]
  5. GOLDWASSER E., WHITE W. F., TAYLOR K. B. Further purification of sheep plasma erythropoietin. Biochim Biophys Acta. 1962 Nov 5;64:487–496. doi: 10.1016/0006-3002(62)90306-2. [DOI] [PubMed] [Google Scholar]
  6. Iscove N. N., Guilbert L. J., Weyman C. 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. 1980 Mar;126(1):121–126. doi: 10.1016/0014-4827(80)90476-0. [DOI] [PubMed] [Google Scholar]
  7. JACOBSON L. O., GOLDWASSER E., GURNEY C. W., FRIED W., PLZAK L. Studies of erythropoietin: the hormone regulating red cell production. Ann N Y Acad Sci. 1959 Jun 25;77:551–573. doi: 10.1111/j.1749-6632.1959.tb36925.x. [DOI] [PubMed] [Google Scholar]
  8. Jelkmann W., Bauer C. Demonstration of high levels of erythropoietin in rat kidneys following hypoxic hypoxia. Pflugers Arch. 1981 Nov;392(1):34–39. doi: 10.1007/BF00584579. [DOI] [PubMed] [Google Scholar]
  9. Kanamaru A., Okamoto T., Hara H., Nagai K. Developmental changes in erythropoietin responsiveness of late erythroid precursors in mouse hemopoietic organs. Dev Biol. 1982 Jul;92(1):221–226. doi: 10.1016/0012-1606(82)90166-x. [DOI] [PubMed] [Google Scholar]
  10. Kurtz A., Jelkmann W., Bauer C. A new candidate for the regulation of erythropoiesis. Insulin-like growth factor I. FEBS Lett. 1982 Nov 22;149(1):105–108. doi: 10.1016/0014-5793(82)81081-8. [DOI] [PubMed] [Google Scholar]
  11. Kurtz A., Jelkmann W., Bauer C. Insulin stimulates erythroid colony formation independently of erythropoietin. Br J Haematol. 1983 Feb;53(2):311–316. doi: 10.1111/j.1365-2141.1983.tb02025.x. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Laubli U. K., Baier W., Binz H., Celio M. R., Humbel R. E. Monoclonal antibodies directed to human insulin-like growth factor I (IGF I). Use for radioimmunoassay and immunopurification of IGF. FEBS Lett. 1982 Nov 22;149(1):109–112. doi: 10.1016/0014-5793(82)81082-x. [DOI] [PubMed] [Google Scholar]
  14. Lenoir D., Honegger P. Insulin-like growth factor I (IGF I) stimulates DNA synthesis in fetal rat brain cell cultures. Brain Res. 1983 Apr;283(2-3):205–213. doi: 10.1016/0165-3806(83)90177-3. [DOI] [PubMed] [Google Scholar]
  15. Matoth Y., Zaizov R. Regulation of erythropoiesis in the fetal rat. Isr J Med Sci. 1971 Jul-Aug;7(7):839–845. [PubMed] [Google Scholar]
  16. Perdue J. F. The role of somatomedin/insulin-like growth factors and their receptors in skeletal growth and fetal development: a mini-review. Prog Clin Biol Res. 1983;132B:405–413. [PubMed] [Google Scholar]
  17. Philipps A. F., Widness J. A., Garcia J. F., Raye J. R., Schwartz R. Erythropoietin elevation in the chronically hyperglycemic fetal lamb. Proc Soc Exp Biol Med. 1982 May;170(1):42–47. doi: 10.3181/00379727-170-41394. [DOI] [PubMed] [Google Scholar]
  18. Rinderknecht E., Humbel R. E. Primary structure of human insulin-like growth factor II. FEBS Lett. 1978 May 15;89(2):283–286. doi: 10.1016/0014-5793(78)80237-3. [DOI] [PubMed] [Google Scholar]
  19. Rinderknecht E., Humbel R. E. The amino acid sequence of human insulin-like growth factor I and its structural homology with proinsulin. J Biol Chem. 1978 Apr 25;253(8):2769–2776. [PubMed] [Google Scholar]
  20. Schoenle E., Zapf J., Humbel R. E., Froesch E. R. Insulin-like growth factor I stimulates growth in hypophysectomized rats. Nature. 1982 Mar 18;296(5854):252–253. doi: 10.1038/296252a0. [DOI] [PubMed] [Google Scholar]
  21. Svoboda M. E., Van Wyk J. J., Klapper D. G., Fellows R. E., Grissom F. E., Schlueter R. J. Purification of somatomedin-C from human plasma: chemical and biological properties, partial sequence analysis, and relationship to other somatomedins. Biochemistry. 1980 Feb 19;19(4):790–797. doi: 10.1021/bi00545a027. [DOI] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Widness J. A., Susa J. B., Garcia J. F., Singer D. B., Sehgal P., Oh W., Schwartz R., Schwartz H. C. Increased erythropoiesis and elevated erythropoietin in infants born to diabetic mothers and in hyperinsulinemic rhesus fetuses. J Clin Invest. 1981 Mar;67(3):637–642. doi: 10.1172/JCI110078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zapf J., Froesch E. R., Humbel R. E. The insulin-like growth factors (IGF) of human serum: chemical and biological characterization and aspects of their possible physiological role. Curr Top Cell Regul. 1981;19:257–309. doi: 10.1016/b978-0-12-152819-5.50024-5. [DOI] [PubMed] [Google Scholar]
  25. Zapf J., Morell B., Walter H., Laron Z., Froesch E. R. Serum levels of insulin-like growth factor (IGF) and its carrier protein in various metabolic disorders. Acta Endocrinol (Copenh) 1980 Dec;95(4):505–517. doi: 10.1530/acta.0.0950505. [DOI] [PubMed] [Google Scholar]
  26. Zapf J., Walter H., Froesch E. R. Radioimmunological determination of insulinlike growth factors I and II in normal subjects and in patients with growth disorders and extrapancreatic tumor hypoglycemia. J Clin Invest. 1981 Nov;68(5):1321–1330. doi: 10.1172/JCI110379. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES