, Volume 80, Issue 6, pp 547–552 | Cite as

Innermembrane association of three mitochondrial β-oxidation enzymes revealed by immunoelectron microscopic technique

  • S. Yokota
  • T. Hashimoto


Ultrastructural localization of three mitochondrial β-oxidation enzymes, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase in rat liver was studied by a post-embedding immunocytochemical technique. Rat liver was fixed by perfusion. Vibratome sections (100 μm thick) were embedded in Lowicryl K4M. Ultrathin sections were separately incubated with antibody to each enzyme, followed by protein A-gold complex. Gold particles representing the antigenic sites for all enzymes examined were confined exclusively to mitochondria of hepatocytes and other sinus-lining cells. Peroxisomes were consistently negative for the immunolabelling. In the mitochondria the gold particles were localized in the matrical side of inner membrane. The control experiments confirmed the specificity of the immunolabelling. The results firstly indicate that the mitochondrial β-oxidation enzymes are present in the matrix of mitochondria and associated with the inner membrane.


Public Health Gold Control Experiment Gold Particle Ultrathin Section 
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  1. Bendayan M, Shore GC (1982) Immunocytochemical localization of mitochondrial proteins in the rat hepatocyte. J Histochem Cytochem 30:139–147Google Scholar
  2. Carlemalm E, Garavito RM, Villiger W (1982) Resin development for electron microscopy and analysis of embedding at low temperature. J Microsc 126:123–143Google Scholar
  3. Debeer LJ, Mannaerts GP (1983) The mitochondrial and peroxisomal pathways of fatty acid oxidation in rat liver. Diabete Metab 9:134–140Google Scholar
  4. Furuta S, Miyazawa S, Osumi T, Hashimoto T, Ui N (1980) Properties of mitochondrial and peroxisomal enoyl-CoA hydratases from rat liver. J Biochem (Tokyo) 88:1059–1070Google Scholar
  5. Horisberger M (1979) Evaluation of colloidal gold as a cytochemical marker for transmission and scanning electron microscopy. Biol Cell 36:253–258Google Scholar
  6. Lazarow PB, de Duve C (1976) A fatty acid oxidizing system in rat liver peroxisomes: Enhancement by clofibrate; a hypolidemic drug. Proc Natl Acad Sci USA 73:2043–2046Google Scholar
  7. Miyazawa S, Osumi T, Hashimoto T (1980) The presence of a new 3-ketoacyl-CoA thiolase in rat liver peroxisomes. Eur J Biochem 103:589–596Google Scholar
  8. Osumi T, Hashimoto T (1980) Purification and properties of mitochondrial and peroxisomal 3-hydroxyacyl-CoA dehydrogenase from rat liver. Arch Biochem Biophys 203:372–383Google Scholar
  9. Roth J (1982) The protein A-gold (pAg) technique. A quantitative and qualitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry. Academic Press, New York, pp 108–133Google Scholar
  10. Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90:533–536Google Scholar
  11. Weibel ED, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30:23–38Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • S. Yokota
    • 1
  • T. Hashimoto
    • 2
  1. 1.Department of AnatomyYamanashi Medical SchoolYamanashiJapan
  2. 2.Department of BiochemistryShinshu University School of MedicineNaganoJapan

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