Glutathione S-transferases in the organ of Corti of the rat: enzymatic activity, subunit composition and immunohistochemical localization.
Glutathione S-transferases (GSTs), a family of ubiquitous cytosolic isozymes, catalyze the detoxification of electrophilic substrates with reduced glutathione and participate in intracellular binding and transport of lipophilic substances. This study measured GST activity biochemically in the inner ear of the rat; determined the isozyme profile by Western blotting; and identified, immunohistochemically, the distribution of the mu and pi class GSTs in the organ of Corti. GST enzymatic activity in inner ear tissues ranged from 117 to 348 nmoles glutathione converted/min/mg protein, values somewhat higher than those found in brain (130) and much lower than in liver (1011). Of the GST isoforms, the pi class (identified by antibodies against the Yp subunit) was most prominent, the mu class (Yb1 subunit) clearly evident while the alpha class (Y(a) subunit) was barely detectable on Western blots. Immunocytochemical analysis showed differential distribution of the Yb1 and Yp subunits. The Yb1 subunit was present in the sensory cells, while supporting cells were not specifically stained. At the subcellular level, the isozyme was localized in the apical zones of inner (IHCs) and outer hair cells (OHCs) close to the cuticular plate. The extent of staining, however, varied between OHCs and IHCs. In the OHCs, staining appeared in discrete spots in the apical areas only, whereas in IHCs staining extended further towards the center of the cells. The Yp subunit was mainly localized to Deiters cell processes and pillar cells. Both Yb1 and Yp colocalized with tubulin-specific antibody. The functional significance of GST in the cochlear receptor cells is speculative. However, a role analogous to that in other tissues (detoxification, prostaglandin synthesis) can be assumed. In addition, an association of GST with the microtubule system is possible based on immunohistochemical colocalization with tubulin.
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Kresge Hearing Research Institute, Department of Otolaryngology, University of Michigan, Ann Arbor 48109-0506.
Altschuler
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Schacht
J
J
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D005982:Glutathione Transferase / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D006199:Hair Cells, Auditory, Inner / Q000201:enzymology*; D018072:Hair Cells, Auditory, Outer / Q000201:enzymology*; D007527:Isoenzymes; D008099:Liver / Q000201:enzymology; D008297:Male; D008856:Microscopy, Fluorescence; D009925:Organ of Corti / Q000201:enzymology*; D016613:Plastic Embedding; D051381:Rats; D017208:Rats, Wistar
D007527:Isoenzymes; D005982:Glutathione Transferase