Dupont E, Zhao H F, Rhéaume E, Simard J, Luu-The V, Labrie F, Pelletier G
Medical Research Council Group in Molecular Endocrinology, CHUL Research Centre, Quebec, Canada.
Endocrinology. 1990 Sep;127(3):1394-403. doi: 10.1210/endo-127-3-1394.
The enzyme complex delta 5-3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta HSD) is involved in the biosynthesis of all classes of steroids, namely glucocorticoids, mineralocorticoids, progesterone, and sex steroids. To obtain information on the precise localization of 3 beta HSD in rat gonads and adrenal glands, two complementary cytochemical techniques were used; immunocytochemical localization was achieved with antibodies developed against purified human placental 3 beta HSD, while 3 beta HSD mRNA localization was achieved by in situ hybridization performed with a recently cloned rat 3 beta HSD cDNA. In the testis, specific immunostaining was restricted to the cytoplasm of the interstitial cells, while by in situ hybridization, specific silver grains were also seen over the interstitial cells. In the ovary, immunostaining was found in the cytoplasm of cells of the corpus luteum and theca interna, while the granulosa cells of the follicles showed no positive reaction. By in situ hybridization, a specific hybridization signal was observed over granulosa cells of the corpus luteum, which are mainly responsible for progesterone secretion, and to a lesser extent over theca interna cells, known for their role in secreting C19 androgens. In the adrenals, the three zones of the cortex were equally immunolabeled, whereas no staining could be detected in the medulla. Similarly, by in situ hybridization, silver grains were located over the zona glomerulosa, fasciculata, and reticularis, while no specific autoradiographic reaction could be observed on the chromaffin cells of the medulla. The present study provides new information about the precise cellular localization of 3 beta HSD in the adrenal glands and gonads in the rat, thus providing useful information about the site of action of 3 beta HSD, especially in the gonads. Moreover, the approaches used for localization studies, especially quantitative in situ hybridization, should provide a useful tool for assessing the role of hormones on 3 beta HSD expression in the different compartments of the gonads and adrenal glands.
δ5 - 3β - 羟基类固醇脱氢酶/δ5 - δ4异构酶（3βHSD）复合酶参与所有类固醇的生物合成，即糖皮质激素、盐皮质激素、孕酮和性激素。为了获取3βHSD在大鼠性腺和肾上腺中精确位置的信息，使用了两种互补的细胞化学技术；用针对纯化的人胎盘3βHSD制备的抗体进行免疫细胞化学定位，而通过用最近克隆的大鼠3βHSD cDNA进行原位杂交来实现3βHSD mRNA定位。在睾丸中，特异性免疫染色局限于间质细胞的细胞质，而通过原位杂交，在间质细胞上也可见特异性银颗粒。在卵巢中，黄体和卵泡内膜细胞的细胞质中有免疫染色，而卵泡的颗粒细胞未显示阳性反应。通过原位杂交，在主要负责孕酮分泌的黄体颗粒细胞上观察到特异性杂交信号，在较小程度上在以分泌C19雄激素作用而闻名的卵泡内膜细胞上也观察到。在肾上腺中，皮质的三个带均有同等程度的免疫标记，而在髓质中未检测到染色。同样，通过原位杂交，银颗粒位于球状带、束状带和网状带，而在髓质的嗜铬细胞上未观察到特异性放射自显影反应。本研究提供了关于大鼠肾上腺和性腺中3βHSD精确细胞定位的新信息，从而提供了关于3βHSD作用位点的有用信息，特别是在性腺中。此外，用于定位研究的方法，尤其是定量原位杂交，应为评估激素对性腺和肾上腺不同区域中3βHSD表达的作用提供有用工具。
