Departments of Molecular and Integrative Physiology and Internal Medicine (J.R., W.E.R.), University of Michigan Medical School, Ann Arbor, Michigan 48109; Departments of Physiology (J.R., T.W., W.E.R.) and Surgery (T.D.M.), Georgia Regents University, Augusta, Georgia 30912; and Department of Pathology (Y.N., H.S.), Tohoku University School of Medicine, Sendai 980-8579, Japan.
J Clin Endocrinol Metab. 2014 Mar;99(3):E518-27. doi: 10.1210/jc.2013-3198. Epub 2013 Jan 1.
The human adrenal zona fasciculata (ZF) and zona reticularis (ZR) are responsible for the production of cortisol and 19-carbon steroids (often called adrenal androgens), respectively. However, the gene profiles and exact molecular mechanisms leading to the functional phenotype of the ZF and ZR are still not clearly defined. In the present study, we identified the transcripts that are differentially expressed in the ZF and ZR.
The objective of the study was to compare the transcriptome profiles of ZF and ZR.
ZF and ZR were microdissected from 10 human adrenals. Total RNA was extracted from 10 ZF/ZR pairs and hybridized to Illumina microarray chips. The 10 most differentially expressed transcripts were studied with quantitative RT-PCR (qPCR). Immunohistochemistry was also performed on four zone-specific genes.
Microarray results demonstrated that only 347 transcripts of the 47 231 were significantly different by 2-fold or greater in the ZF and ZR. ZF had 195 transcripts with 2-fold or greater increase compared with its paired ZR, whereas ZR was found to have 152 transcripts with 2-fold or greater higher expression than in ZF. Microarray and qPCR analysis of transcripts encoding steroidogenic enzymes (n = 10) demonstrated that only 3β-hydroxysteroid dehydrogenase, steroid sulfotransferase, type 5 17β-hydroxysteroid dehydrogenase, and cytochrome b5 were significantly different. Immunohistochemistry and qPCR studies confirmed that the ZF had an increased expression of lymphoid enhancer-binding factor 1 and nephroblastoma overexpressed, whereas ZR showed an increased expression of solute carrier family 27 (fatty acid transporter) (SLC27A2), member 2 and TSPAN12 (tetraspanin 12) CONCLUSION: Microarray revealed several novel candidate genes for elucidating the molecular mechanisms governing the ZF and ZR, thereby increasing our understanding of the functional zonation of these two adrenocortical zones.
人体肾上腺束状带（ZF）和网状带（ZR）分别负责产生皮质醇和 19 碳甾体（通常称为肾上腺雄激素）。然而，导致 ZF 和 ZR 功能表型的基因谱和确切分子机制尚不清楚。在本研究中，我们鉴定了在 ZF 和 ZR 中差异表达的转录本。
本研究旨在比较 ZF 和 ZR 的转录组谱。
从 10 个人肾上腺中微分离 ZF 和 ZR。从 10 对 ZF/ZR 中提取总 RNA 并杂交至 Illumina 微阵列芯片。用定量 RT-PCR（qPCR）研究了 10 个差异最显著的转录本。还对四个区特异性基因进行了免疫组织化学染色。
微阵列结果表明，ZF 和 ZR 中只有 47231 个转录本中的 347 个差异在 2 倍或更高。ZF 中有 195 个转录本的表达水平是其配对 ZR 的 2 倍或更高，而 ZR 中发现有 152 个转录本的表达水平是 ZF 的 2 倍或更高。对编码类固醇生成酶的转录本（n = 10）的微阵列和 qPCR 分析表明，只有 3β-羟甾脱氢酶、类固醇硫酸转移酶、5 型 17β-羟甾脱氢酶和细胞色素 b5 有显著差异。免疫组织化学和 qPCR 研究证实，ZF 表达增强了淋巴增强结合因子 1 和肾母细胞瘤过表达，而 ZR 则表现出溶质载体家族 27（脂肪酸转运体）（SLC27A2）成员 2 和 TSPAN12（四跨膜蛋白 12）的表达增加。
微阵列揭示了几个新的候选基因，用于阐明调节 ZF 和 ZR 的分子机制，从而增加了我们对这两个肾上腺皮质区功能分区的理解。
