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Fine localization of serine: pyruvate aminotransferase in rat hepatocytes revealed by a post-embedding immunocytochemical technique

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Summary

Immunocytochemical localization of serine: pyruvate aminotransferase (SPT) in rat hepatocytes was studied using a protien A-gold technique. Rat liver was fixed by perfusion. Vibratome sections (100 μm thick) of the liver were embedded in Epon or Lowicryl K4M. Ultrathin sections were incubated with antiSPT, followed by protein A-gold complex. Gold particles representing the antigenic sites for SPT were seen in three subcellular compartments, peroxisomes, mitochondria, and cytoplasm. In the control experiments the specificity of the immunolabelling was confirmed. Quantitative analysis of the labelling density showed that main subcellular compartments containing SPT are mitochondria and peroxisomes. In addition, the gold particles distributing in the cytoplasm were 16%–29% of the total labelling. The result indicated that the cytoplasm also contains SPT with a low density.

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References

  • Carlemalm E, Garavito RM, Villiger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126:123–143

    Google Scholar 

  • Cheung GP, Cotropia JP, Sallach HJ (1969) The effects of dietary protein on the hepatic enzymes of serine metabolism in the rabbit. Arch Biochem Biophys 129:672–682

    Google Scholar 

  • Fahimi HD, Kalmbach P, Stegmeier K, Stork H (1980) Comparison between the effects of clofibrate and bezafibrate upon the ultrastructure of rat heart and liver. In: Greten H, Cang PD, Schettler G (eds) Lipoproteins and coronary heart disease. New aspects in the diagnosis and therapy of disorders of lipid metabolism. G Witzstrock, New York, pp 64–75

    Google Scholar 

  • Fahimi HD, Reinecke A, Sujatta M, Yokota S, Özel M (1982) The short- and long-term effects of bezafibrate in the rat. Ann NY Acad Sci USA 386:111–133

    Google Scholar 

  • Fukushima M, Aihara Y, Ichiyama A (1978) Immunochemical studies on induction of rat liver mitochondrial serine: pyruvate aminotransferase by glucagon. J Biol Chem 253:1187–1194

    Google Scholar 

  • Hess R, Stäubli W, Reis W (1965) Nature of the hepatomegalic effect produced by ethyl-chlorophenoxy-isobutyrate in the rat. Nature 208:856–858

    Google Scholar 

  • Lazarow PB (1977) Three hypolipidemic drugs increase hepatic palmitoyl-coenzyme A oxidation in the rat. Science 197:580–581

    Google Scholar 

  • Lazarow PB, de Duve C (1976) A fatty acy-CoA oxidizing system in rat liver peroxisomes: enhancement by clofibrate, a hypolipidemic drug. Proc Natl Acad Sci USA 73:2043–2046

    Google Scholar 

  • Lazarow PB, Shio H, Leroy-Honyet MA (1982) Specificity in the action of hypolipidemic drugs: increase of peroxisomal β-oxidation largely dissociated from hepatomegaly and peroxisome proliferation in rat. J Lipid Res 23:317–326

    Google Scholar 

  • Noguchi T, Okuno E, Takada Y, Minatogawa Y, Okai K, Kido R (1978) Subcellular distribution of pyruvate (glyoxylate) aminotransferases in rat liver. Biochem J 170:173–175

    Google Scholar 

  • Oda T, Yanagisawa M, Ichiyama A (1982) Induction of serine:pyruvate aminotransferase in rat liver organelles by glucagon and high protein diet. J Biochem 91:219–232

    Google Scholar 

  • Reddy JK, Krishnakantha TP (1975) Hepatic peroxisome proliferation: induction by two novel compounds structurally unrelated to clofibrate. Science 190:787–789

    Google Scholar 

  • Roswell EV, Snell K, Carnie JA, Al-Tai AH (1969) Liver L-alanine-glyoxylate and L-serine-pyruvate aminotrasferase activities: An apparent association with gluconeogenesis. Biochem J 115:1071–1073

    Google Scholar 

  • Roswell EV, El-Tai AH, Carnie JA, Roswell KV (1973) Increased liver L-serine-pyruvate aminotransferase activity under gluconeogenic conditions. Biochem J 134:349–351

    Google Scholar 

  • 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 108–133

    Google Scholar 

  • Sallach HJ, Sanborn TA, Bruin WJ (1972) Dietary and hormonal regulation of hepatic biosynthetic and catabolic enzymes of serine metabolism in rats. Endocrinology 91:1054–1063

    Google Scholar 

  • Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90:533–536

    Google Scholar 

  • Stathis EC, Fabrikanos A (1958) Preparation of colloidal gold. Chem Ind (London) 27:860–861

    Google Scholar 

  • Walsh DA, Sallach HJ (1966) Comparative studies on the pathways for serine biosynthesis in animal tissues. J Biol Chem 241:4068–4076

    Google Scholar 

  • Weibel ED, Kistler GS, Scherle WF (1966) Practical stereological methods for morphological cytology. J Cell Biol 30:23–38

    Google Scholar 

  • Yokota S, Oda T (1983) Immunoelectron microscopic localization of serine: pyruvate aminotransferase in rat cosinophile leukocytes. Histochemistry 78:417–424

    Google Scholar 

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Yokota, S., Oda, T. Fine localization of serine: pyruvate aminotransferase in rat hepatocytes revealed by a post-embedding immunocytochemical technique. Histochemistry 80, 591–595 (1984). https://doi.org/10.1007/BF00553722

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