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Single Cell Cytochemistry Illustrated by the Demonstration of Glucose-6-Phosphate Dehydrogenase Deficiency in Erythrocytes

  • Anna L. Peters
  • Cornelis J. F. van NoordenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1560)

Abstract

Cytochemistry is the discipline that is applied to visualize specific molecules in individual cells and has become an essential tool in life sciences. Immunocytochemistry was developed in the sixties of last century and is the most frequently used cytochemical application. However, metabolic mapping is the oldest cytochemical approach to localize activity of specific enzymes, but in the last decades of the previous century and the first decade of the present century it almost became obsolete. The popularity of this approach revived in the past few years. Metabolism gained interest as player in chronic and complex diseases such as cancer, diabetes, neurodegenerative diseases, and vascular diseases and both enzyme cytochemistry and metabolic mapping have become important tools in life sciences.

In this chapter, we present glucose-6-phosphate dehydrogenase (G6PD) deficiency, the most prevalent enzyme deficiency worldwide, to illustrate recent developments in enzyme cytochemistry or metabolic mapping. The first assays which were developed quantified enzyme activity but were unreliable for single cell evaluation. The field has expanded with the development of cytochemical single cell assays and DNA testing. Still, all assays—from the earliest developed tests up to the most recently developed tests—have their place in investigations on G6PD activity. Recently, nanoscopy has become available for light and fluorescence microscopy at the nanoscale. For nanoscopy, cytochemistry is an essential tool to visualize intracellular molecular processes. The ultimate goal in the coming years will be nanoscopy of living cells so that the molecular dynamics can be studied. Cytochemistry will undoubtedly play a critical role in these developments.

Key words

Cytochemistry History Metabolic mapping Glucose-6-phosphate dehydrogenase deficiency 

References

  1. 1.
    Heilemann M (2010) Fluorescence microscopy beyond the diffraction limit. J Biotechnol 149(4):243–251. doi: 10.1016/j.jbiotec.2010.03.012 CrossRefPubMedGoogle Scholar
  2. 2.
    Galbraith CG, Galbraith JA (2011) Super-resolution microscopy at a glance. J Cell Sci 124(Pt 10):1607–1611. doi: 10.1242/jcs.080085 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Hell SW (2009) Microscopy and its focal switch. Nat Methods 6(1):24–32. doi: 10.1038/nmeth.1291 CrossRefPubMedGoogle Scholar
  4. 4.
    Gomori G (1939) Microtechnical demonstration of phosphatase in tissue sections. Exp Biol Med 42(1):23–26CrossRefGoogle Scholar
  5. 5.
    Van Noorden CJ (2010) Imaging enzymes at work: metabolic mapping by enzyme histochemistry. J Histochem Cytochem 58(6):481–497. doi: 10.1369/jhc.2010.955518 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Van Noorden CJ (2014) Metabolic mapping by (quantitative) enzyme hystochemistry. In: McManus L, Mitchell R (eds) Pathobiology of human disease. Elsevier, San Diego, CA, pp 3760–3774CrossRefGoogle Scholar
  7. 7.
    Bleeker FE, Atai NA, Lamba S et al (2010) The prognostic IDH1(R132) mutation is associated with reduced NADP+-dependent IDH activity in glioblastoma. Acta Neuropathol 119(4):487–494. doi: 10.1007/s00401-010-0645-6 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Koehler A, Van Noorden CJ (2003) Reduced nicotinamide adenine dinucleotide phosphate and the higher incidence of pollution-induced liver cancer in female flounder. Environm Toxicol Chem 22(11):2703–2710CrossRefGoogle Scholar
  9. 9.
    Luzzatto L (2006) Glucose 6-phosphate dehydrogenase deficiency: from genotype to phenotype. Haematologica 91(10):1303–1306PubMedGoogle Scholar
  10. 10.
    Howes RE, Piel FB, Patil AP et al (2012) G6PD deficiency prevalence and estimates of affected populations in malaria endemic countries: a geostatistical model-based map. PLoS Med 9(11), e1001339. doi: 10.1371/journal.pmed.1001339 CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Nkhoma ET, Poole C, Vannappagari V et al (2009) The global prevalence of glucose-6-phosphate dehydrogenase deficiency: a systematic review and meta-analysis. Blood Cell Mol Dis 42(3):267–278. doi: 10.1016/j.bcmd.2008.12.005 CrossRefGoogle Scholar
  12. 12.
    Peters AL, Van Noorden CJ (2009) Glucose-6-phosphate dehydrogenase deficiency and malaria: cytochemical detection of heterozygous G6PD deficiency in women. J Histochem Cytochem 57(11):1003–1011. doi: 10.1369/jhc.2009.953828 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Guindo A, Fairhurst RM, Doumbo OK et al (2007) X-Linked G6PD deficiency protects hemizygous males but not heterozygous females against severe malaria. PLoS Med 4(3), e66. doi: 10.1371/journal.pmed.0040066 CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Cappellini MD, Fiorelli G (2008) Glucose-6-phosphate dehydrogenase deficiency. Lancet 371(9606):64–74. doi: 10.1016/S0140-6736(08)60073-2 CrossRefPubMedGoogle Scholar
  15. 15.
    Meletis J, Konstantopoulos K (2004) Favism—from the “avoid fava beans” of Pythagoras to the present. Haematology 7(1):17–21Google Scholar
  16. 16.
    Beutler E (2008) Glucose-6-phosphate dehydrogenase deficiency: a historical perspective. Blood 111(1):16–24. doi: 10.1182/blood-2007-04-077412 CrossRefPubMedGoogle Scholar
  17. 17.
    Dern R, Weinstein IM, Leroy G et al (1954) The hemolytic effect of primaquine. I. The localization of the drug-induced hemolytic defect in primaqulne-sensitive individuals. J Lab Clin Med 43(2):303–309PubMedGoogle Scholar
  18. 18.
    Crosby WH (1956) Favism in Sardinia (newsletter). Blood 11(1):91–92Google Scholar
  19. 19.
    Sansone G, Segni G (1958) New aspects of the biochemical alterations in the erythrocytes of patients with favism; almost complete absence of glucose-6-phosphate dehydrogenase. Boll Soc Ital Biol Sper 34(7):327PubMedGoogle Scholar
  20. 20.
    Lyon MF (1961) Gene action in the X-chromosome of the mouse (Mus musculus L.). Nature 190:372–373CrossRefPubMedGoogle Scholar
  21. 21.
    Rattazzi MC (1968) Glucose 6-phosphate dehydrogenase from human erythrocytes: molecular weight determination by gel filtration. Biochem Biophys Res Commun 31(1):16–24CrossRefPubMedGoogle Scholar
  22. 22.
    Luzzatto L, Battistuzzi G (1985) Glucose-6-phosphate dehydrogenase. Adv Hum Genet 14:217–329PubMedGoogle Scholar
  23. 23.
    Wrigley NG, Heather JV, Bonsignore A et al (1972) Human erythrocyte glucose 6-phosphate dehydrogenase: electron microscope studies on structure and interconversion of tetramers, dimers and monomers. J Mol Biol 68(3):483–499CrossRefPubMedGoogle Scholar
  24. 24.
    Marks PA, Johnson AB, Hirschberg E (1958) Effect of age on the enzyme activity in erythrocytes. Proc Natl Acad Sci U S A 44(6):529–536CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    WHO Working Group (1989) Glucose-6-phosphate dehydrogenase deficiency. B World Health Organ 67(6):601–611Google Scholar
  26. 26.
    Beutler E (1994) G6PD deficiency. Blood 84(11):3613–3636PubMedGoogle Scholar
  27. 27.
    World Health Organization (1967) Standardization of procedures for the study of glucose-6-phosphate dehydrogenase. Report of a WHO Scientific Group. W Health Organ Tech Rep Ser 366:1–53Google Scholar
  28. 28.
    LaRue N, Kahn M, Murray M et al (2014) Comparison of quantitative and qualitative tests for glucose-6-phosphate dehydrogenase deficiency. Am J Trop Med Hyg 91(4):854–861. doi: 10.4269/ajtmh.14-0194 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Nantakomol D, Paul R, Palasuwan A et al (2013) Evaluation of the phenotypic test and genetic analysis in the detection of glucose-6-phosphate dehydrogenase deficiency. Malar J 12:289. doi: 10.1186/1475-2875-12-289 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Jonges GN, Hagen H, Van Noorden CJ et al (1989) Comparison between the chromate inhibition test and a cytochemical method for the determination of glucose-6-phosphate dehydrogenase deficiency in erythrocytes. Clin Chim Acta 181(2):135–141CrossRefPubMedGoogle Scholar
  31. 31.
    Wolf BH, Weening RS, Schutgens RB et al (1987) Detection of glucose-6-phosphate dehydrogenase deficiency in erythrocytes: a spectrophotometric assay and a fluorescent spot test compared with a cytochemical method. Clin Chim Acta 168(2):129–136CrossRefPubMedGoogle Scholar
  32. 32.
    Van Noorden CJ, Vogels IM, James J et al (1982) A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. I. Optimalization of the staining procedure. Histochemistry 75(4):493–506CrossRefPubMedGoogle Scholar
  33. 33.
    Van Noorden CJ, Vogels IM (1985) A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. II. Further improvements of the staining procedure and some observations with glucose-6-phosphate dehydrogenase deficiency. Br J Haematol 60(1):57–63CrossRefPubMedGoogle Scholar
  34. 34.
    Persico MG, Viglietto G, Martini G et al (1986) Isolation of human glucose-6-phosphate dehydrogenase (G6PD) cDNA clones: primary structure of the protein and unusual 5′ non-coding region. Nucleic Acids Res 14(6):2511–2522CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Trask BJ, Massa H, Kenwrick S et al (1991) Mapping of human chromosome Xq28 by two-color fluorescence in situ hybridization of DNA sequences to interphase cell nuclei. Am J Hum Genet 48(1):1–15PubMedPubMedCentralGoogle Scholar
  36. 36.
    Martini G, Toniolo D, Vulliamy T et al (1986) Structural analysis of the X-linked gene encoding human glucose 6-phosphate dehydrogenase. EMBO J 5(8):1849–1855PubMedPubMedCentralGoogle Scholar
  37. 37.
    Minucci A, Moradkhani K, Hwang MJ et al (2012) Glucose-6-phosphate dehydrogenase (G6PD) mutations database: review of the “old” and update of the new mutations. Blood Cell Mol Dis 48(3):154–165. doi: 10.1016/j.bcmd.2012.01.001 CrossRefGoogle Scholar
  38. 38.
    Von Seidlein L, Auburn S, Espino F et al (2013) Review of key knowledge gaps in glucose-6-phosphate dehydrogenase deficiency detection with regard to the safe clinical deployment of 8-aminoquinoline treatment regimens: a workshop report. Malar J 12:112. doi: 10.1186/1475-2875-12-112 CrossRefGoogle Scholar
  39. 39.
    Van Noorden CJ, Dolbeare F, Aten J (1989) Flow cytofluorometric analysis of enzyme reactions based on quenching of fluorescence by the final reaction product: detection of glucose-6-phosphate dehydrogenase deficiency in human erythrocytes. J Histochem Cytochem 37(9):1313–1318CrossRefPubMedGoogle Scholar
  40. 40.
    Shah SS, Diakite SA, Traore K et al (2012) A novel cytofluorometric assay for the detection and quantification of glucose-6-phosphate dehydrogenase deficiency. Sci Rep 2:299. doi: 10.1038/srep00299 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Anna L. Peters
    • 1
  • Cornelis J. F. van Noorden
    • 2
    Email author
  1. 1.Department of Intensive CareAcademic Medical CenterAmsterdamThe Netherlands
  2. 2.Department of Cell Biology and HistologyAcademic Medical CenterAmsterdamThe Netherlands

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