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Nuklearmedizin 1974; 13(01): 72-84
DOI: 10.1055/s-0038-1624845
Originalarbeiten — Original Articles
Schattauer GmbH

Isolation of the 67Gallium Accumulating Fraction in Normal Rat Liver

Die Isolation der 67Gallium anreichernden Fraktion der normalen Rattenleber
K. Hierholzer
1   Aus dem Institut für Klinische Physiologie, Klinikum Steglitz, Freie Universität Berlin (Direktor: Prof. Dr. K. Hierholzer)
,
K. zum Winkel
2   der Strahlenklinik und Poliklinik der Freien Universität Berlin, Klinikum Westend (Direktor: Prof. Dr. K. zum Winkel)
,
E. Aulbert
3   with technical assistance of M. Hristowa and B. Wilhelm
,
U. Haubold
1   Aus dem Institut für Klinische Physiologie, Klinikum Steglitz, Freie Universität Berlin (Direktor: Prof. Dr. K. Hierholzer)
› Author Affiliations
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Publication History

Received:00 January 1974

Publication Date:
12 January 2018 (online)

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Summary

Subcellular 67Gallium distribution was investigated in normal rat liver after intravenous injection. By differential centrifugation and density gradient centrifugation 67Gallium accumulating bodies were isolated and identified as lysosomes by enzyme determination and electron microscopy. 67Gallium enrichment in this fraction was 23-fold. Using the isolated 67Gallium accumulating lysosomes the binding state of the isotope inside the lysosomes was studied. 67Gallium was found to be associated with the soluble fraction of lysosomes.

Die subzelluläre Verteilung von 67Gallium wurde in der normalen Rattenleber nach intravenöser Tracer-Injektion untersucht. Mit Hilfe der Differentialzentrifugation sowie der Dichtegradientenzentrifugation konnte eine 67Ga-akkumulierende Fraktion isoliert werden, die eine 23fache 67Ga-Anreicherung gegenüber dem Gesamthomogenat aufwies. Die 67Ga-anreichernden Partikel wurden enzymatisch sowie elektronenoptisch als Lysosomen identifiziert.

An den isolierten 67Ga-reichen Lysosomen wurde die intralysosomale Verteilung des Isotops näher untersucht. Es zeigte sich dabei, daß das 67Gallium nicht an die lysosomale Membran oder an Strukturproteine gebunden vorliegt, sondern sich in der löslichen Fraktion der Lysosomen befindet.

* N.V. Philips Duphar Cyclotron and Isotope Laboratories, Petten, Netherlands. 1 ml of the original solution contained 1.25-5 mg sodium citrate, 9.0 mg benzyl alkohol, 1-2 mCi 67Ga (first day) carrier-free, phenol red as indicator.


 

  • References

  • 1 Edwards C. L, Hayes R. L. Tumor-Scanning with 67Ga-Citrate. J. Nucl. Med 1969; 10: 103.
    PubMedGoogle Scholar
  • 2 Edwards C. L, Hayes R. L, Nelson B. M, Tehranian N. Clinical Investigation of 67Ga Tumor Scanning. J. Nucl. Med 1970; 11: 316.
    Google Scholar
  • 3 Edwards C. L, Hayes R. L. Scanning Malignant Neoplasms with 67Gallium. J. Amer. med. Ass 1970; 212: 1182.
    CrossrefGoogle Scholar
  • 4 Hayes R. L, Nelson B. M, Swartzendruber D. C, Carlton J. E, Byrd B. L. 67Gallium Localization in Rat and Mouse Tumors. Science 1970; 167: 289.
    CrossrefPubMedGoogle Scholar
  • 5 Higasi T, Nakayama Y. Clinical Evaluation of 67Ga-Citrate Scanning. J. Nucl. Med 1972; 13: 196.
    PubMedGoogle Scholar
  • 6 Palermo F, Patrese P. Utilizzazione Del Radiogallio (Ga-67-Citrato) Nell'Individuazione Di Lesioni Neoplastiche. Reperti Clinici E Possibilità Diagnostiche. La Ricerca Clin. Lab 1972; 2: 538.
    Google Scholar
  • 7 Haubold U, Koppenhagen K, Schmid-Burgk F. Tumorszintigraphie mit 67Ga-Citrat. Deutscher Röntgenkongress 1970. (Hug. O. ed.) Thieme; Stuttgart: 1970: 127.
    Google Scholar
  • 8 Haubold U, Aulbert E. 67Gallium as a Tumor-Scanning Agent. Clinical and Physiological Aspects. In: Medical Radioisotope Scintigraphy 1972.(International Atomic Energy Agency ed.), Vol. II: 553. Vienna: 1973
    Google Scholar
  • 9 Orii H. Tumor scanning with gallium (67Ga) and its mechanism studied in rats. Strahlentherapie 1972; 144 (02) 192.
    Google Scholar
  • 10 Ito Y, Okuyama S, Sato K, Takahashi K, Sato T, Kanno I. 67Ga Tumor Scanning and its Mechanisms Studied in Rabbits. Radiology 1971; 100 (02) 357.
    CrossrefPubMedGoogle Scholar
  • 11 Swartzendruber D. C, Nelson B. M, Hayes R. L. 67Gallium localization in lysosomal-like granules of leukemic and nonleukemic murine tissues. J. Nat. Cancer Inst 1971; 46: 941.
    PubMedGoogle Scholar
  • 12 Widnell C. C, Tata J. R. A procedure for the isolation of enzymically active rat-liver nuclei. Biochem. J 1964; 92: 313.
    CrossrefPubMedGoogle Scholar
  • 13 Emmelot P, Bos C. J, Benedetti E. L Ph. Rümke . Studies on plasma membranes. III. Mg2+-ATPase, (Na+K+-Mg2+)-ATPase and 5'-nucleotidase activity of plasma membranes isolated from rat liver. Biochim. Biophys. Acta 1964; 90: 126.
    CrossrefPubMedGoogle Scholar
  • 14 Ebel H, De Santo N. G, Hierholzer K. Plasma Cell Membranes of the Rat Kidney. Pflügers Arch 1971; 324: 1.
    CrossrefPubMedGoogle Scholar
  • 15 Lowry O. H, Rosenbrough N. J, Farr A. L, Randall R. J. Protein measurement with the Folin phenol reagent. J. Biol. Chem 1951; 193: 265.
    PubMedGoogle Scholar
  • 16 Fiske C. H, Subbarow Y. The colorimetric determination of phosphorus. J. Biol. Chem 1925; 66: 375.
    Google Scholar
  • 17 Swanson M. A. Phosphatases of liver. Glucose-6-phosphatase. J. Biol. Chem 1950; 184: 647.
    PubMedGoogle Scholar
  • 18 Hübscher G, West G. R. Specific assays of some phosphatases in subcellular fractions of small intestinal mucosa. Nature 1965; 205: 799.
    CrossrefPubMedGoogle Scholar
  • 19 Andersch M. A, Szczypinski A. J. Use of p-nitrophenyl-phosphate as substrate in determination of serum acid phosphatase. Amer. J. clin. Path 1947; 17: 571.
    Google Scholar
  • 20 Fishman W. H, Lerner F. A Method for Estimating Serum Acid Phosphatase of Prostatic Origin. J. Biol. Chem 1953; 200: 89.
    PubMedGoogle Scholar
  • 21 Talalay P, Fishman W. H, Huggins C. Chromogenic Substrates II. Phenolphthalein Glucuronic Acid as Substrate for the Assay of Glucuronidase Activity. J. Biol. Chem 1946; 166: 757.
    PubMedGoogle Scholar
  • 22 Stahl P. D, Touster O. ß-Glucuronidase of Rat Liver Lysosomes. J. Biol. Chem 1971; 246: 5398.
    PubMedGoogle Scholar
  • 23 Green D. E, Mii S, Kohout P. M. Studies on the terminal electron transport system 1. Succinic dehydrogenase. J. Biol. Chem 1955; 217: 551.
    PubMedGoogle Scholar
  • 24 Seubert W. Methoden der enzymatischen Analyse. Bergmeyer H. U. ed Verlag Chemie; Weinheim: 1962: 433.
    Google Scholar
  • 25 Bücher Th W, Pette Luh D. Einfache und zusammengesetzte optische Tests mit Pyridinnucleotiden in: Hoppe-Seyler/Thierfelder: Handbuch der physiologischund pathologisch-chemischen Analyse. Lang K, Lehnartz eds E. 10th Ed. Vol. VI Springer; Berlin-Göttingen-Heidelberg: 1964: 292.
    Google Scholar
  • 26 Chvapil M, Ryan J. N, Brada Z. Effects of Selected Chelating Agents and Metals on the Stability of Liver Lysosomes. Biochem. Pharm 1972; 21: 1097.
    CrossrefPubMedGoogle Scholar
  • 27 De Duve C. General Properties of Lysosomes. In: Ciba Foundation Symposium: Lysosomes. De Reuck A. V, Cameron M. P. Eds Little, Brown & Co; Boston: 1963: 1-35.
    Google Scholar
  • 28 De Duve C, Pressmann B. C, Gianetto R, Wattiaux R, Appelmans F. Tissue Fractionation Studies 6. Intracellular Distribution Patterns of Enzymes in Rat-Liver Tissue. Biochem 1955; 60: 604.
    CrossrefPubMedGoogle Scholar
  • 29 Wattiaux R, Wibo M, Baudhuin P. Influence of the Injection of Triton WR- 1339 on the Properties of Rat-Liver Lysosomes. In: Ciba Foundation Symposium: Lysosomes. De Reuck A.V, Cameron M. P. Eds Little, Brown & Co; Boston: 1963: 176-196.
    Google Scholar
  • 30 Deckner K, Becker G, Langowski U, Schwering H, Hornung G, Schmidt C. G. Die subcelluläre Bindung von 67Gallium in Ascites-Tumorzellen. Z. Krebsforsch 1971; 76: 293.
    CrossrefPubMedGoogle Scholar
  • 31 Tappel A. L, Sawant P. L, Shibko S. Lysosomes: Distribution in Animais, Hydrolytic Capacity and other Properties. In: Ciba Foundation Symposium: Lysosomes. De Reuck A. V, Cameron M. P. eds Little, Brown & Co; Boston: 1963: 78-108.
    Google Scholar
  • 32 Bitensky L. The Reversible Activation of Lysosomes in Normal Cells and the Effects of Pathological Conditions. In: Ciba Foundation Symposium: Lysosomes. De Reuck A. V, Cameron M. P. eds Little, Brown & Co; Boston: 1963: 362-375
    Google Scholar
  • 33 Allison A. C. The Role of Lysosomes in Pathology. Proc. R. Soc. Med 1966; 59: 867.
    PubMedGoogle Scholar
  • 34 Dianzani M. U. Lysosome Changes in Liver Injury. In: Ciba Foundation Symposium: Lysosomes. De Reuck A. V, Cameron M. P. eds Little, Brown & Co; Boston: 1963: 335-352
    Google Scholar
  • 35 Swartzendruber D. C, Byrd B. L, Hayes R. L, Nelson B, Tyndall R. L. Preferential Localization of 67Gallium Citrate in Tissues of Leukemic Mice. J. Nat. Cancer Inst 1970; 44 (03) 695.
    PubMedGoogle Scholar