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Glucose-6-phosphate dehydrogenase is enriched in oligodendrocytes of the rat spinal cord

Enzyme histochemical and immunocytochemical studies

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Abstract

Glucose-6-phosphate dehydrogenase (G6PD) was localized in rat spinal cord by catalytic enzyme histochemistry and immunocytochemistry. G6PD detected by either method was shown to be strongly enriched in cell bodies and processes of oligodendrocytes, whereas in the compact myelin G6PD was not detected. The enzyme histochemical procedure for the demonstration of G6PD was also adapted for microphotometric measurements of G6PD activity in the spinal cord white matter. There was a linear relationship between G6PD activity and section thickness up to 14 μm and between G6PD activity and reaction time up to 5–6 min as demonstrated by kinetic and end-point measurements. Significantly lower activities were measured in endpoint measurements than in kinetic measurements because of formazan loss during rinsing. Methoxyphenazine methosulphate as an exogenous electron carrier and sodium azide as a blocker of the respiratory chain significantly increased the demonstrable G6PD activity. The K m was 0.62 mM and the V max 3 μmol glucose-6-phosphate/cm3 wet tissue and per min at 25°C. It is concluded that G6PD in oligodendrocytes may be important for the generation of NADPH required for lipid biosynthesis related to myelogenesis, and reduction of glutathione required for protection of membrane sulphydryl groups.

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References

  • Appel SH, Parrot BL (1970) Hexose monophosphate pathway in synapses. J Neurochem 17:1619–1626

    Google Scholar 

  • Beutler E, Morrison M (1967) Localization and characteristics of hexose 6-phosphate dehydrogenase (glucose dehydrogenase). J Biol Chem 242:5289–5293

    Google Scholar 

  • Bignami A, Eng LF, Dahl D, Uyeda CT (1972) Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res 43:429–435

    Google Scholar 

  • Burnette WN (1981) Western blotting: electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radionated protein A. Anal Biochem 112:195–203

    Google Scholar 

  • Butcher RG (1978) The measurement in tissue sections fo the two formazans derived from nitroblue tetrazolium in dehydrogenase reactions. Histochem J 10:739–744

    Google Scholar 

  • Butcher RG, Dawson AL, Knaab SA, Gahan PB (1980) Dehydrogenase activity and loss of formazan from tissue sections. Histochem J 12:591–598

    Google Scholar 

  • Butcher RG, Van Noorden CJF (1985) Reaction rate studies of glucose-6-phosphate dehydrogenase activity in sections of rat liver using four tetrazolium salts. Histochem J 17:993–1008

    Google Scholar 

  • Cammer W, Snyder DS, Zimmerman TR, Jr., Farooq M, Norton WT (1982) Glycerol phosphate dehydrogenase, glucose-6-phosphate dehydrogenase, and lactate dehydrogenase: activities in oligodendrocytes, neurons, astrocytes, and myelin isolated from developing rat brains. J Neurochem 38:360–367

    Google Scholar 

  • Cammer W, Downing M, Clarke W, Schenkmann JB (1991) Immuno-cytochemical staining of the RLM6 form of cytochrome P-450 in oligodendrocytes and myelin of rat brain. J Histochem Cytochem 39:1089–1094

    Google Scholar 

  • Cho SW, Joshi JG (1990) Characterization of glucose-6-phosphate dehydrogenase isozymes from human and pig brain. Neuroscience 38:819–828

    Google Scholar 

  • Deutsch J (1983) Glucose-6-phosphate dehydrogenase. In: Bergmeyer HH (ed) Methods of enzymatic analysis. Verlag Chemie, Weinheim Deerfield Beach Basel, pp 190–197

    Google Scholar 

  • Drenckhahn D, Franz H (1986) Identification of actin-, α -actinin-, and vinculin-containing plaques at the lateral membrane of epithelial cells. J Cell Biol 102:1843–1852

    Google Scholar 

  • Drenckhahn D, Mannherz HG (1983) Distribution of actin and the actin-associated proteins myosin, tropomyosin, alpha-actinin, vinculin, and villin in rat and bovine exocrine glands. Eur J Cell Biol 30:167–176

    Google Scholar 

  • Drenckhahn D, Hofmann H-D, Mannherz HG (1983) Evidence for the association of villin with core filaments and rootlets of intestinal epithelial microvilli. Cell Tissue Res 228:409–414

    Google Scholar 

  • Drenckhahn D, Jöns T, Schmitz F (1993) Production of polyclonal antibodies against proteins and peptides. Methods Cell Biol 37:7–56

    Google Scholar 

  • Friede RL, Fleming LM, Knoller M (1963) A comparative mapping of enzymes involved in hexosemonophosphate shunt and citric acid cycle in the brain. J Neurochem 10:263–277

    Google Scholar 

  • Hawkins RA, Mans AM (1983) Intermediary metabolism of carbohydrates and other fuels. In: Lajtha A (ed) Handbook of neurochemistry, vol 3, 2nd edn. Plenum Press, New York, pp 259–294

    Google Scholar 

  • Hino Y, Minakami S (1982) Hexose-6-phosphate dehydrogenase of rat liver microsomes. J Biol Chem 257:2563–2568

    Google Scholar 

  • Jonges, GN, Van Noorden CJF (1989) In situ kinetic parameters of glucose-6-phosphate dehydrogenase and phosphogluconate dehydrogenase in different areas of the rat liver acinus. Histochem J 21:585–594

    Google Scholar 

  • Jöns T, Drenckhahn D (1992) Identification of the binding interface involved in linkage of cytoskeletal protein 4.1 to the erythrocyte anion exchanger. EMBO J 11:2863–2867

    Google Scholar 

  • Kalina M, Plapinger RE, Hoshino Y, Seligman AM (1972) Nonosmiophilic tetrazolium salts that yield osmiophilic, lipophobic formazans for ultrastructural localization of dehydrogenase activity. J Histochem Cytochem 20:685–695

    Google Scholar 

  • Kugler P (1982) Quantitative histochemistry of the lysosomal dipeptidyl aminopeptidase II in the proximal tubule of the rat kidney. Histochemistry 76:557–566

    Google Scholar 

  • Kugler P (1988) Quantitative enzyme histochemistry in the brain. Histochemistry 90:99–107

    Google Scholar 

  • Kugler P (1990a) Microphotometric determination of enzymes in brain sections. I. Hexokinase. Histochemistry 93:295–298

    Google Scholar 

  • Kugler P (1990b) Microphotometric determination of enzymes in brain sections. III. Glutamate dehydrogenase. Histochemistry 93:537–540

    Google Scholar 

  • Kugler P (1991) Microphotometric determination of enzymes in brain sections. V. Glycerophosphate dehydrogenase. Histochemistry 95:579–583

    Google Scholar 

  • Kugler P, Baier G (1990) Microphotometric determination of enzymes in brain sections. II. GABA transaminase. Histochemistry 93:501–505

    Google Scholar 

  • Kugler P, Vogel S (1991) Microphotometric determination of enzymes in brain sections. IV. Isocitrate dehydrogenases. Histochemistry 95:629–633

    Google Scholar 

  • Kugler P, Vogel S, Gehm M (1988) Quantitative succinate dehydrogenase histochemistry in the hippocampus of aged rats. Histochemistry 88:299–307

    Google Scholar 

  • Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666

    Google Scholar 

  • Major HD, Hampton JC, Rosario B (1961) A simple method for removing the resin from epoxy-embedded tissue. J Biophys Biochem Cytol 9:909–910

    Google Scholar 

  • Mandula B, Srivastava SK, Beutler E (1970) Hexose-6-phosphatedehydrogenase: distribution in rat tissues and effects of diet, age and steroids. Arch Biochem Biophys 141:155–161

    Google Scholar 

  • McDougal DB Jr., Schulz DW, Passonneau JV, Clark JR, Reynolds MA, Lowry OH (1961) Quantitative studies of white matter. J Gen Physiol 44:487–498

    Google Scholar 

  • Nogae I, Johnston M (1990) Glucose-6-phosphate dehydrogenase. Gene 96:161–169

    Google Scholar 

  • Ouchterlony Ö, Nilsson L-A (1978) Immunodiffusion and immuno-electrophoresis. In: Weir DM (ed) Handbook of experimental immunology. Blackwell Scientific Publications, Oxford

    Google Scholar 

  • Sprinkle TJ (1989) 2′,3′-Cyclic nucleotide 3′-phosphodiesterase, an oligodendrocyte-Schwann cell and myelin-associated enzyme of the nervous system. CRC Crit Rev Neurobiol 4:235

    Google Scholar 

  • Tanahashi K, Hori SH (1980) Immunohistochemical localization of hexose 6-phosphate dehydrogenase in various organs of the rat. J Histochem Cytochem 28:1175–1182

    Google Scholar 

  • Van Noorden CJF (1984) Histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase. Prog Histochem Cytochem Vol 15 Nr 4

  • Van Noorden CJF, Kooij A, Vogels IMC, Frederiks WM (1985) On the nature of the ‘nothing dehydrogenase’ reaction. Histochem J 17:1111–1118

    Google Scholar 

  • Van Noorden CJF, Vogels IMC (1989) Polyvinyl alcohol and other tissue protectants in enzyme histochemistry: a consumer's guide. Histochem J 21:373–379

    Google Scholar 

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  1. Institute of Anatomy, University of Würzburg, Koellikerstrasse 6, D-97070, Würzburg, Germany

    P. Kugler

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Kugler, P. Glucose-6-phosphate dehydrogenase is enriched in oligodendrocytes of the rat spinal cord. Histochemistry 101, 143–153 (1994). https://doi.org/10.1007/BF00269361

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