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转录组分析揭示了大鼠肾上腺球状带和束状带中差异表达的转录本。

Transcriptome analysis reveals differentially expressed transcripts in rat adrenal zona glomerulosa and zona fasciculata.

机构信息

Department of Physiology, Georgia Health Sciences University, Augusta, Georgia 30912, USA.

出版信息

Endocrinology. 2012 Apr;153(4):1755-63. doi: 10.1210/en.2011-1915. Epub 2012 Feb 28.

Abstract

In mammals, aldosterone is produced in the zona glomerulosa (zG), the outermost layer of the adrenal cortex, whereas glucocorticoids are produced in adjacent zona fasciculata (zF). However, the cellular mechanisms controlling the zonal development and the differential hormone production (i.e. functional zonation) are poorly understood. To explore the mechanisms, we defined zone-specific transcripts in this study. Eleven-week-old male rats were used and adrenal tissues were collected from zG and zF using laser-capture microdissection. RNA was isolated, biotin labeled, amplified, and hybridized to Illumina microarray chips. The microarray data were compared by fold change calculations. In zG, 235 transcripts showed more than a 2-fold up-regulation compared to zF with statistical significance. Similarly, 231 transcripts showed up-regulation in zF. The microarray findings were validated using quantitative RT-PCR and immunohistochemical staining on selected transcripts, including Cyp11b2 (zG/zF: 214.2x), Rgs4 (68.4x), Smoc2 (49.3x), and Mia1 (43.1x) in zG as well as Ddah1 (zF/zG 16.2x), Cidea (15.5x), Frzb (9.5x), and Hsd11b2 (8.3x) in zF. The lists of transcripts obtained in the current study will be an invaluable tool for the elucidation of cellular mechanisms leading to zG and zF functional zonation.

摘要

在哺乳动物中,醛固酮由肾上腺皮质的最外层——球状带(zG)产生,而糖皮质激素则由相邻的束状带(zF)产生。然而,控制区域发育和激素产生(即功能分区)的细胞机制尚不清楚。为了探索这些机制,我们在本研究中定义了区域特异性转录本。使用 11 周龄雄性大鼠,使用激光捕获显微解剖术从 zG 和 zF 采集肾上腺组织。分离 RNA,用生物素标记、扩增,并与 Illumina 微阵列芯片杂交。通过折叠变化计算比较微阵列数据。在 zG 中,与 zF 相比,有 235 个转录本的上调倍数超过 2 倍,且具有统计学意义。同样,有 231 个转录本在 zF 中上调。使用定量 RT-PCR 和选定转录本的免疫组织化学染色对微阵列发现进行了验证,包括 Cyp11b2(zG/zF:214.2x)、Rgs4(68.4x)、Smoc2(49.3x)和 Mia1(43.1x)在 zG 中,以及 Ddah1(zF/zG:16.2x)、Cidea(15.5x)、Frzb(9.5x)和 Hsd11b2(8.3x)在 zF 中。本研究中获得的转录本列表将是阐明导致 zG 和 zF 功能分区的细胞机制的宝贵工具。

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