Single Cell Cytochemistry Illustrated by the Demonstration of Glucose-6-Phosphate Dehydrogenase Deficiency in Erythrocytes.
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.
Department of Intensive Care, Academic Medical Center, Meibergdreef 9, Amsterdam, 1105AZ, The Netherlands.
Anna L
AL
Peters
Department of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15, 1105AZ, Amsterdam, The Netherlands. c.j.vannoorden@amc.uva.nl.
Cornelis J F
CJ
van Noorden
D004789:Enzyme Activation; D004912:Erythrocytes / Q000378:metabolism*; D005820:Genetic Testing / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000175:diagnosis / Q000201:enzymology* / Q000235:genetics*; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D010084:Oxidation-Reduction; D010427:Pentose Phosphate Pathway; D059010:Single-Cell Analysis / Q000379:methods; D013053:Spectrophotometry
D005954:Glucosephosphate Dehydrogenase
Cytochemistry; Glucose-6-phosphate dehydrogenase deficiency; History; Metabolic mapping