Light and electron microscopic immunocytochemistry on the localization of 3?-hydroxysteroid dehydrogenase/isomerase in the bovine adrenal cortical cells

Ishimura, Kazunori; Yoshinaga-Hirabayashi, Tomoko; Fujita, Hisao; Hiroko, Ishii-Ohba; Inano, H.; Tamaoki, Bun‐Ichi

doi:10.1007/bf00496581

Cited by 21 publications

(13 citation statements)

References 24 publications

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“…In bovine and rat adrenal cortex, 3 -HSD has been shown to be associated with both the mitochondrial and microsomal fractions (Cherradi et al 1993, Sauer et al 1994. By immunoelectron microscopy, 3 -HSD immunoreaction was found in the ER in bovine adrenal cortex (Ishimura et al 1988, Ishimura & Fujita 1997. Recently, Cherradi et al (1997) using immunoelectron microscopy have reported that 3 -HSD immunoreactivity could be detected in both ER and mitochondria, the highest density being observed in the ER, as shown in the present study.…”

Section: Discussionsupporting

confidence: 85%

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Immunoelectron microscopic localization of three key steroidogenic enzymes (cytochrome P450(scc), 3 beta-hydroxysteroid dehydrogenase and cytochrome P450(c17)) in rat adrenal cortex and gonads

Pelletier¹,

Li²,

Luu‐The³

et al. 2001

Journal of Endocrinology

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The biosynthesis of steroid hormones in endocrine steroidsecreting glands results from a series of successive steps involving both cytochrome P450 enzymes, which are mixed-function oxidases, and steroid dehydrogenases. So far, the subcellular distribution of steroidogenic enzymes has been mostly studied following subcellular fractionation, performed in placenta and adrenal cortex. In order to determine in situ the intracellular distribution of some steroidogenic enzymes, we have investigated the ultrastructural localization of the three key enzymes: P450 side chain cleavage (scc) which converts cholesterol to pregnenolone; 3 -hydroxysteroid dehydrogenase (3 -HSD) which catalyzes the conversion of 3 -hydroxy-5-ene steroids to 3-oxo-4-ene steroids (progesterone and androstenedione); and P450 c17 which is responsible for the transformation of C 21 into C 19 steroids (dehydroepiandrosterone and androstenedione). Immunogold labeling was used to localize the enzymes in rat adrenal cortex and gonads. The tissues were fixed in 1% glutaraldehyde and 3% paraformaldehyde and included in LR gold resin. In the adrenal cortex, both P450 scc and 3 -HSD immunoreactivities were detected in the reticular, fascicular and glomerular zones. P450 scc was exclusively found in large mitochondria. In contrast, 3 -HSD antigenic sites were mostly observed in the endoplasmic reticulum (ER) with some gold particles overlying crista and outer membranes of the mitochondria. P450 c17 could not be detected in adrenocortical cells. In the testis, the three enzymes were only found in Leydig cells. Immunolabeling for P450 scc and 3 -HSD was restricted to mitochondria, while P450 c17 immunoreactivity was exclusively observed in ER. In the ovary, P450 scc and 3 -HSD immunoreactivities were found in granulosa, theca interna and corpus luteum cells. The subcellular localization of the two enzymes was very similar to that observed in adrenocortical cells. P450 c17 could also be detected in theca interna cells of large developing and mature follicles. As observed in Leydig cells, P450 c17 immunolabeling could only be found in the ER. These results indicate that in different endocrine steroid-secreting cells P450 scc , 3 -HSD and P450 c17 have the same association with cytoplasmic organelles (with the exception of 3 -HSD in Leydig cells), suggesting similar intracellular pathways for biosynthesis of steroid hormones.

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Section: Discussionsupporting

confidence: 85%

“…The ultrastructural localization of steroidogenic enzymes has, so far, not been extensively investigated and the few reports on the subject are restricted to the adrenal cortex (Ishimura et al 1988, Shinzawa et al 1988, Whitnall et al 1993, Cherradi et al 1997, Ishimura & Fujita 1997.…”

Section: Introductionmentioning

confidence: 99%

Immunoelectron microscopic localization of three key steroidogenic enzymes (cytochrome P450(scc), 3 beta-hydroxysteroid dehydrogenase and cytochrome P450(c17)) in rat adrenal cortex and gonads

Pelletier¹,

Li²,

Luu‐The³

et al. 2001

Journal of Endocrinology

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“…At the electron microscopic level, association of the enzyme with the smooth endoplasmic reticulum in the adrenal cortical cells has been confirmed (Ishimura et al 1988). In peripheral organs where there is no typical steroid-secreting cell, the subcellular distribution of 3 -HSD has not so far been reported.…”

Section: Discussionmentioning

confidence: 97%

“…In bovine and rat adrenal cortex, 3 -HSD activity has been shown to be associated with both the mitochondrial and microsomal fractions (Cherradi et al 1993, Sauer et al 1994. By immunoelectron microscopy, 3 -HSD immunoreactivity was found to be associated only with the smooth endoplasmic reticulum (Ishimura et al 1988) in bovine adrenal cortex.…”

Section: Introductionmentioning

confidence: 93%

“…In fact, the ultrastructural localization of steroidogenic enzymes obtained by immunoelectron microscopy has not so far been extensively investigated and the few reports on the subject are related to classical steroidogenic glands such as the testis, ovary and adrenal cortex (Ishimura et al 1988, Shinzawa et al 1988, Whitnall et al 1993. We have recently developed antibodies against type 5 17 -HSD and shown by immunocytochemistry that this enzyme could be detected in reproductive organs, including the prostate, uterus and mammary gland (El-Alfy et al 1999.…”

Section: Introductionmentioning

confidence: 99%

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Immunoelectron microscopic localization of 3beta-hydroxysteroid dehydrogenase and type 5 17beta-hydroxysteroid dehydrogenase in the human prostate and mammary gland

Pelletier

Luu‐The²,

El‐Alfy³

et al. 2001

Journal of Molecular Endocrinology

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The subcellular distribution of steroidogenic enzymes has so far been studied mostly in classical endocrine glands and in the placenta. In the peripheral intracrine organs which synthesize sex steroids there is no indication about the organelles which contain the enzymes involved in steroid biosynthesis. We have thus investigated the subcellular localization of two enzymes involved in the production of sex steroids, namely 3 -hydroxysteroid dehydrogenase (3 -HSD) and type 5 17 -hydroxysteroid dehydrogenase (17 -HSD). Using specific antibodies to these enzymes, we conducted immunoelectron microscopic studies in two peripheral tissues, namely the human prostate and mammary gland. In the prostate, immunolabelling for both 3 -HSD and type 5 17 -HSD was detected in the basal cells of the tube-alveoli as well as in fibroblasts and endothelial cells lining the blood vessels. In all the labelled cell types, the gold particles were distributed throughout the cytoplasm. No obvious association with any specific organelle could be observed, although some concentration of gold particles was occasionally found over bundles of microfilaments. In mammary gland sections immunolabelled for 3 -HSD or type 5 17 -HSD localization, labelling was observed in the cytoplasm of the secretory epithelial cells in both the acini and terminal ducts. Immunolabelling was also found in the endothelial cells as well as in fibroblasts in stroma and blood vessels. The gold particles were not detected over any organelles, except with the occasional accumulation of gold particles over microfilaments. The present data on the localization of two steroidogenic enzymes leading to the synthesis of testosterone indicate that these enzymes are located not only in epithelial cells but also in stromal and endothelial cells in both tissues studied. The absence of any association of the enzymes with membrane-bound organelles appears as a common finding in the reactive cell types of two peripheral tissues.

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Light and electron microscopic immunohistochemistry of the localization of adrenal steroidogenic enzymes

Ishimura

Fujita

1997

Microsc. Res. Tech.

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Light and electron microscopic immunocytochemistry on the localization of 3?-hydroxysteroid dehydrogenase/isomerase in the bovine adrenal cortical cells

Cited by 21 publications

References 24 publications

Immunoelectron microscopic localization of three key steroidogenic enzymes (cytochrome P450(scc), 3 beta-hydroxysteroid dehydrogenase and cytochrome P450(c17)) in rat adrenal cortex and gonads

Immunoelectron microscopic localization of three key steroidogenic enzymes (cytochrome P450(scc), 3 beta-hydroxysteroid dehydrogenase and cytochrome P450(c17)) in rat adrenal cortex and gonads

Immunoelectron microscopic localization of 3beta-hydroxysteroid dehydrogenase and type 5 17beta-hydroxysteroid dehydrogenase in the human prostate and mammary gland

Light and electron microscopic immunohistochemistry of the localization of adrenal steroidogenic enzymes

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