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Zusammenfassung Unter Anwendung histochemisch-elektrophoretischer Technik wurde versucht, die Aktivitat der Sorbitdehydrogenase in der Rattenniere zu erfassen. Im histochemischen Test ist gegenwartig eine adaquate Erfassung der Aktivitat der leicht diffusiblen Sorbitdehydrogenase in nativen Schnitten unter Anwendung der Membraninkubationstechnik in Gegenwart von PMS and KCN moglich. Der starke uniforme Reaktionsausfall in den Glomerula ist hinsichtlich seiner Spezifitat noch nicht abgeklart. In semidunnen Schnitten glutaraldehydvorfixierter, inkubierter, Epon -eingebetteter Kryostatdickschnitte ist das Reaktionsprodukt im Nierenglomerulum vorwiegend in den mesangialen Zellen lokalisiert. Nach der Mikropolyacrylamidgel-Gradienten-Elektrophorese des Uberstandes der Nierenrinde Bowie isolierter Glornerula and Inkubation der Gelzylinder in den gleichen histochemischen Medien besteht Anhalt fur das Vorliegen von 2 Sorbitdehydrogenase-Fraktionen. Die Ergebnisse werden im Hinblick auf ihre Spezifitat besprochen.

We have investigated the distribution of sorbitol dehydrogenase by comparative indicator- and immunohistochemical technique in rat kidney. In this connection we have tested some parameters which influence the histochemical demonstration of SoDH-activity by tetrazolium salt indicator technique (e.g. prefixation of tissue sections, elution behaviour of SoDH, influence of different dialysis membranes, composition of incubating medium). According these studies, the enzyme was demonstrated in native cryostate sections by membrane incubating method. For the immunohistochemical demonstration of SoDH specific antibodies were prepared against this enzyme in rabbits, and were used for indirect immunofluorescence method and for the unlabelled antibody enzyme (PAP) technique. A good agreement of sorbitol dehydrogenase localization was obtained with all the tested methods, but also some discrepancies was observed in enzyme topic (e.g. glomeruli). The results are discussed under methodical aspects in connection with the specifity of the used methods to localize the enzyme.

We have investigated the distribution of sorbitol dehydrogenase by comparative indicator- and immunohistochemical technique in rat kidney. In this connection we have tested some parameters which influence the histochemical demonstration of SoDH-activity by tetrazolium salt indicator technique (e.g. prefixation of tissue sections, elution behaviour of SoDH, influence of different dialysis membranes, composition of incubating medium). According these studies, the enzyme was demonstrated in native cryostate sections by membrane incubating method. For the immunohistochemical demonstration of SoDH specific antibodies were prepared against this enzyme in rabbits, and were used for indirect immunofluorescence method and for the unlabelled antibody enzyme (PAP) technique. A good agreement of sorbitol dehydrogenase localization was obtained with all the tested methods, but also some discrepancies was observed in enzyme topic (e.g. glomeruli). The results are discussed under methodical aspects in connection with the specifity of the used methods to localize the enzyme.

We have investigated the localization and demonstration of sorbitol dehydrogenase activity in the rat kidney by comparative histochemical-electrophoretical technique. Under histochemical conditions an adequate demonstration of the activity of the sorbitol dehydrogenase in native sections is possible by membrane incubating technique in presence of PMS and KCN. In the glomerulum the strong and uniform reaction with regard to the specifity is not clear. In the glomerula of semithin sections from glutaraldehyde fixed, incubated and Epon-embedded cryostate sections the reaction product is localized predominantly in the maesangium cells. By micropolyacrylamidgel gradient electrophoresis of the supernatant from the kidney cortex and of isolated glomerula 2 sorbitol dehydrogenase fractions were found. The results are discussed in connection with the specifity of the reaction.