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Quantitative immunocytochemical studies on differential induction of serine

Pyruvate aminotransferase in mitochondria and peroxisomes of rat liver cells by administration of glucagon or di-(2-ethylhexyl)phthalate

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Summary

Differential induction of serine: pyruvate aminotransferase (SPT) in rat liver parenchymal cells by administration of glucagon or di-(2-ethylhexyl)phthalate (DEHP) was studied using post-embedding immunocytochemical techniques and morphometric methods. Two groups of rats were fasted for 5 days and daily received peritoneal injection of glucagon (300 μg/100 g) or physiological saline. Another two groups of rats were fed on laboratory chow with or without 2% DEHP for 2 weeks. Livers were perfusionfixed, cut into tissue sections (50–100 μ), and processed to cytochemistry for catalase, immunocytochemistry for SPT, and conventional procedures for electron microscopy. The morphometric analysis showed that glucagon injection has negligible effect on the volume and numerical density and mean diameter of peroxisomes, whereas volume density of mitochondria was decreased by 25%. By DEHP administration peroxisomes were about 3-fold increased in the volume and numerical density. Mitochondria was increased about 40% in the numerical density, but unchanged in the volume density. Light and electron microscopic immunocytochemistry demonstrated that glucagon injection exclusively enhanced mitochondrial SPT, whereas DEHP administration exclusively induced in peroxisomal SPT. Quantitative analysis showed that by the glucagon injection, the labeling density of mitochondria was increased about 4-fold, but that of peroxisomes was 1.6 times as much as control, while by DEHP administration, the labeling density of peroxisomes was enhanced about 3-fold but that of mitochondria was decreased by 13%. The results clearly indicate that glucagon induces mitochondrial SPT, whereas peroxisome proliferator, DEHP induces peroxisomal SPT.

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References

  • Avrameas S, Ternynck T (1971) Peroxidase labelled antibody and Fab conjugates with enhanced intracellular penctration. Immunochemistry 8:1175–1179

    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, Reinecke A, Sujatta M, Yokota S, Özel M, Hartig F, Stegmeier K (1982) The short-and long-term effects of bezafibrate in the rat. Ann NY Acad Sci 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 

  • Graham RC, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302

    Google Scholar 

  • Hashimoto T (1982) Individual peroxisomal-oxidation enzymes. Ann NY Acad Sci 386:5–12

    Google Scholar 

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

    Google Scholar 

  • Hoshino J, Robert B, Kroger H (1974) Induction in vivo of rat liver serine: pyruvate aminotransferase by N6,O2′-dibutyryl cyclic AMP and its inhibition by cortisone. Biochim Biophys Acta 338:418–427

    Google Scholar 

  • Kishida Y, Olsen BR, Berg RA, Prockop DJ (1975) Two improved methods for preparing ferritin-protein conjugated for electron microscopy. J Cell Biol 64:331–399

    Google Scholar 

  • Lardy H, Veneziale C, Gabrielli F (1969) Paths of carbon in gluconcogenesis. FEBS Symposium 19:55–62

    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 acyl-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) Specficity in the action of hypolipidemic drugs: increase of peroxisomal-oxidation largely dissociated from hepatomegaly and peroxisome proliferation in rat. J Lipid Res 23:317–326

    Google Scholar 

  • LeHir M, Herzog V, Fahimi HD (1979) Cytochemical detection of catalase with 3,3′-diaminobenzidine. A quantitative reinvestigation of the optimal conditions. Histochemistry 64:51–66

    Google Scholar 

  • Leighton F, Coloma L, Koenig C (1975) Structure, composition, physical properties, and turnover of proliferated peroxisomes. A study of the trophic effects of Su-13437 on rat liver. J Cell Biol 67:281–309

    Google Scholar 

  • Litwin JA, Yokota S, Hashimoto T, Fahimi HD (1984) Light microscopic immunocytochemical demonstration of peroxisomal enzymes in Epon sections. Histochemistry 81:15–22

    Google Scholar 

  • Moody DE, Reddy JK (1976) Morphometric analysis of the ultrastructural changes in rat induced by the peroxisome proliferator sall 42348. J Cell Biol 71:768–780

    Google Scholar 

  • Noguchi T, Okuno E, Kido R (1976) Identity of isoenzyme 1 of histidine-pyruvate aminotransferase with serine-pyruvate aminotransferase. Biochem J 159:607–613

    Google Scholar 

  • Noguchi T, Takada Y, Kido R (1977) Characteristics of hepatic serine-pyruvate aminotransferase in different mammalian species. Biochem J 161:609–614

    Google Scholar 

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

    Google Scholar 

  • Noguchi T, Takada Y, Oota Y (1979) Intraperoxisomal and intramitochondrial localization, and assay of pyruvate (glyoxylate) aminotransferase from rat liver. Hoppe-Seyler's Z Physiol Chem 360:919–927

    Google Scholar 

  • Oda T, Ichiyama A, Miura S, Mori M, Tatibana M (1981) In vitro synthesis of putative precursor of serine: pyruvate aminotransferase of rat liver mitochondria. Biochem Biophys Res Commun 102:568–573

    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 (Tokyo) 91:219–232

    Google Scholar 

  • Oda T, Ichiyama A, Miura S, Mori M (1984) Uptake and processing of serine: pyruvate aminotransferase precursor by rat liver mitochondria in vitro and in vivo. J Biochem (Tokyo) 95:815–824

    Google Scholar 

  • Ohno S, Ohtake N, Fujii Y, Yamabayashi S, Usuda N, Nagata T (1981) Histochemical studies on peroxisomes of rat livers during DEHP administration and after withdrawal on thick sections by means of light microscopy and high voltage electron microscopy. Acta Histochem Cytochem 14:126–142

    Google Scholar 

  • Osumi T, Hashimoto T (1984) The inducible fatty acid oxidation system in mammalian peroxisomes. TIBS 9:317–319

    Google Scholar 

  • Panchenko LF, Popova SV, Antonenkov VD (1982) Inducing effect of clofibrate on alkaline phosphatase and histidine-glyoxylate aminotransferase in rat liver. Experientia 38:433–434

    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 

  • Reddy JK, Lalwani ND, Qureshi SA, Reddy MK, Moehle CM (1984) Induction of hepatic peroxisome proliferation in nonrodent species, including primates. Am J Pathol 114:171–183

    Google Scholar 

  • Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M (1981) The enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671

    Google Scholar 

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

    Google Scholar 

  • Rowsell EV, Al-Tai AH, Carnie AJ (1972) Liver L-serine-pyruvate aminotransferase activity in different animal species. Biochem J 127:21 p

    Google Scholar 

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

    Google Scholar 

  • Rowsell KV, Al-Naama MR, Benett P (1982) The subcellular distribution of rat liver serine-pyruvate aminotransferase. Biochem J 202:483–490

    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 

  • Shindo Y, Osumi T, Hashimoto T (1978) Effects of administration of di-(2-ethylhexyl)phthalate on rat liver mitochondria. Biochem Pharmacol 27:2683–2688

    Google Scholar 

  • Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for electron 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 

  • Staubli W, Schweizer W, Suter J, Weibel ER (1977) The proliferative response of hepatic peroxisomes of neonatal rats to treatment with SU-13 437 (nafenopin). J Cell Biol 74:665–689

    Google Scholar 

  • Takada Y, Noguchi T (1981) Increase in hepatic pyruvate (glyoxylate) aminotransferase activity on administration of clofibrate to the rat. Biochem Pharmacol 30:393–394

    Google Scholar 

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

    Google Scholar 

  • Yanagisawa M, Higashi S, Oda T, Ichiyama A (1983) Properties and possible physiological role of rat liver serine: pyruvate aminotransferase. In: Lennon DLF, Stratman FW, Zahlten RN (eds) Biochemistry of metabolic processes. Elsevier Biomedical, New York, pp 413–426

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Yokota S, Oda T (1985) Immunocytochemical demonstration of serine: pyruvate aminotransferase in peroxisomes and mitochondria of rat kidney. Histochemistry 83:81–85

    Google Scholar 

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  1. Department of Anatomy, Yamanashi Medical School, 409-38, Yamanashi, Japan

    S. Yokota

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Yokota, S. Quantitative immunocytochemical studies on differential induction of serine. Histochemistry 85, 145–155 (1986). https://doi.org/10.1007/BF00491762

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