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In Situ Localization of Transketolase Activity in Epithelial Cells of Different Rat Tissues and Subcellularly in Liver Parenchymal Cells

Joan Boren, Antonio Ramos-Montoya, Klazien S. Bosch, Heleen Vreeling, Ard Jonker, Josep J. Centelles, Marta Cascante and Wilma M. Frederiks

Department of Biochemistry and Molecular Biology, CERQT-Parc Cientific de Barcelona, IDIBAPS, University of Barcelona, Barcelona, Catalonia, Spain (JB,AR-M,JJC,MC), and Department of Cell Biology and Histology, University of Amsterdam, Academical Medical Centre, Amsterdam, The Netherlands (KSB,HV,AJ,WMF)

Correspondence to: Dr. Wilma M. Frederiks, Department of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. E-mail: w.m.frederiks{at}amc.uva.nl

Metabolic mapping of enzyme activities (enzyme histochemistry) is an important tool to understand (patho)physiological functions of enzymes. A new enzyme histochemical method has been developed to detect transketolase activity in situ in various rat tissues and its ultrastructural localization in individual cells. In situ detection of transketolase is important because this multifunctional enzyme has been related with diseases such as cancer, diabetes, Alzheimer's disease, and Wernicke-Korsakoff's syndrome. The proposed method is based on the tetrazolium salt method applied to unfixed cryostat sections in the presence of polyvinyl alcohol. The method appeared to be specific for transketolase activity when the proper control reaction is performed and showed a linear increase of the amount of final reaction product with incubation time. Transketolase activity was studied in liver, small intestine, trachea, tongue, kidney, adrenal gland, and eye. Activity was found in liver parenchyma, epithelium of small intestine, trachea, tongue, proximal tubules of kidney and cornea, and ganglion cells in medulla of adrenal gland. To demonstrate transketolase activity ultrastructurally in liver parenchymal cells, the cupper iron method was used. It was shown that transketolase activity was present in peroxisomes and at membranes of granular endoplasmic reticulum. This ultrastructural localization is similar to that of glucose-6-phosphate dehydrogenase activity, suggesting activity of the pentose phosphate pathway at these sites. It is concluded that the method developed for in situ localization of transketolase activity for light and electron microscopy is specific and allows further investigation of the role of transketolase in (proliferation of) cancer cells and other pathophysiological processes. (J Histochem Cytochem 54:191–199, 2006)

Key Words: transketolase • enzyme histochemistry • electron microscopy • pentose phosphate pathway

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