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Fine structural localization of 3β-hydroxysteroid dehydrogenase in rat corpus luteum

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Synopsis

A procedure for the ultracytochemical demonstration of 3β-hydroxysteroid dehydrogenase (3β-HSD) is described and the results for the localization of this enzyme in the lutein cells of the rat corpus luteum are presented.

The procedure involved pre-fixation with aldehydes and incubation of small blocks. Short fixation (maximum 30 min) in 0.1% glutaraldehyde, 2% depolymerized paraformaldehyde or in 6.25% hydroxyadipaldehyde was found to be the best compromise for the preservation of both 3β-HSD activity and fine structure. The method utilizes 3β-hydroxy-5β-androstan-17-one as substrate, tetranitro blue tetrazolium as a final electron acceptor, and phenazine methosulphate or menadione as an intermediate electron carrier to bypass the NADH2-diaphorase. Control and inhibitor (cyano-ketone) experiments provide evidence for the specificity of the cytochemical reaction.

Results showed that 3β-HSD activity is localized on or in membranes of smooth endoplasmic reticulum, in outer compartments of mitochondria, and possibly within smooth endoplasmic reticulum cisternae and intracristal spaces of mitochondria.

Localization of NADH2-diaphorase activity in the same tissue was also examined. With the ferricyanide techniques, the reaction product was found to be associated mainly with the mitochondria.

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  1. Gönül Bara

    Present address: Department of II. Zoology, Faculty of Science, University of Istanbul, Istanbul, Turkey

Authors and Affiliations

  1. Laboratory of Cellular and Reproductive Biology, Department of Anatomy, 60637, Chicago, Illinois, USA

    Gönül Bara & Winston A. Anderson

  2. the Biomedical Center for Population Research, University of Chicago, 60637, Chicago, Illinois, USA

    Gönül Bara & Winston A. Anderson

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  1. Gönül Bara
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  2. Winston A. Anderson
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Bara, G., Anderson, W.A. Fine structural localization of 3β-hydroxysteroid dehydrogenase in rat corpus luteum. Histochem J 5, 437–449 (1973). https://doi.org/10.1007/BF01012001

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