Abstract
In summary, the data demonstrate that, by the use of repeated injections of an iron sorbitol complex, it is possible to isolate a fraction highly enriched in hydrolytic enzymes (60 times over the homogenate) and in well preserved lysosomes emanating almost entirely from liver parenchymal cells. The advantage of adding fixative to the bottom of the gradient and of using en bloc staining with uranyl acetate is also demonstrated.
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Selected References
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- Arborgh B. A., Glaumann H., Ericsson J. L. Studies on iron loading of rat liver lysosomes. Effects on the liver and distribution and fate of iron. Lab Invest. 1974 May;30(5):664–673. [PubMed] [Google Scholar]
- Coffey J. W., De Duve C. Digestive activity of lysosomes. I. The digestion of proteins by extracts of rat liver lysosomes. J Biol Chem. 1968 Jun 25;243(12):3255–3263. [PubMed] [Google Scholar]
- Coleman R., Michell R. H., Finean J. B., Hawthorne J. N. A purified plasma membrane fraction isolated from rat liver under isotonic conditions. Biochim Biophys Acta. 1967 Sep 9;135(4):573–579. doi: 10.1016/0005-2736(67)90089-2. [DOI] [PubMed] [Google Scholar]
- DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Duve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol. 1966;28:435–492. doi: 10.1146/annurev.ph.28.030166.002251. [DOI] [PubMed] [Google Scholar]
- Drysdale J. W., Munro H. N. Regulation of synthesis and turnover of ferritin in rat liver. J Biol Chem. 1966 Aug 10;241(15):3630–3637. [PubMed] [Google Scholar]
- FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glaumann H., Dallner G. Lipid composition and turnover of rough and smooth microsomal membranes in rat liver. J Lipid Res. 1968 Nov;9(6):720–729. [PubMed] [Google Scholar]
- Henning R., Plattner H., Stoffel W. Nature and localization of acidic groups on lysosomal membranes. Biochim Biophys Acta. 1973 Nov 30;330(1):61–75. doi: 10.1016/0005-2736(73)90284-8. [DOI] [PubMed] [Google Scholar]
- Hopwood D. Some aspects of fixation with glutaraldehyde. A biochemical and histochemical comparison of the effects of formaldehyde and glutaraldehyde fixation on various enzymes and glycogen, with a note on penetration of glutaraldehyde into liver. J Anat. 1967 Jan;101(Pt 1):83–92. [PMC free article] [PubMed] [Google Scholar]
- Leighton F., Poole B., Beaufay H., Baudhuin P., Coffey J. W., Fowler S., De Duve C. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968 May;37(2):482–513. doi: 10.1083/jcb.37.2.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
- NOVIKOFF A. B., BEAUFAY H., DE DUVE C. Electron microscopy of lysosomerich fractions from rat liver. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):179–184. [PMC free article] [PubMed] [Google Scholar]
- Stahn R., Maier K. P., Hannig K. A new method for the preparation of rat liver lysosomes. Separation of cell organelles of rat liver by carrier-free continuous electrophoresis. J Cell Biol. 1970 Sep;46(3):576–591. doi: 10.1083/jcb.46.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weibel E. R., Stäubli W., Gnägi H. R., Hess F. A. Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods, and normal morphometric data for rat liver. J Cell Biol. 1969 Jul;42(1):68–91. doi: 10.1083/jcb.42.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wisse E. An ultrastructural characterization of the endothelial cell in the rat liver sinusoid under normal and various experimental conditions, as a contribution to the distinction between endothelial and Kupffer cells. J Ultrastruct Res. 1972 Mar;38(5):528–562. doi: 10.1016/0022-5320(72)90089-5. [DOI] [PubMed] [Google Scholar]