Department of Molecular and Integrative Physiology, University of Michigan Medical School, Ann Arbor, MI, USA.
Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan Medical School, Ann Arbor, MI, USA.
Mol Cell Endocrinol. 2021 Jun 15;530:111296. doi: 10.1016/j.mce.2021.111296. Epub 2021 Apr 26.
Adequate access to fresh or frozen normal adrenal tissue has been a primary limitation to the enhanced characterization of the adrenal zones via RNA sequencing (RNAseq). Herein, we describe the application of targeted RNAseq to formalin-fixed paraffin-embedded (FFPE) normal adrenal gland specimens. Immunohistochemistry (IHC) was used to visualize and guide the capture of the adrenocortical zones and medulla. Following IHC-based tissue capture and isolation of RNA, high-throughput targeted RNAseq highlighted clear transcriptomic differences and identified differentially expressed genes among the adrenal zones. Our data demonstrate the ability to capture FFPE adrenal zone tissue for targeted transcriptomic analyses. Future comparison of normal adrenal zones will improve our understanding of transcriptomic patterns and help identify potential novel pathways controlling zone-specific steroid production.
充分获取新鲜或冷冻的正常肾上腺组织一直是通过 RNA 测序（RNAseq）增强肾上腺区特征描述的主要限制。在此，我们描述了靶向 RNAseq 在福尔马林固定石蜡包埋（FFPE）正常肾上腺标本中的应用。免疫组织化学（IHC）用于可视化和指导肾上腺皮质区和髓质的捕获。在基于 IHC 的组织捕获和 RNA 分离之后，高通量靶向 RNAseq 突出显示了明显的转录组差异，并鉴定了肾上腺区之间差异表达的基因。我们的数据证明了捕获 FFPE 肾上腺区组织进行靶向转录组分析的能力。未来对正常肾上腺区的比较将提高我们对转录组模式的理解，并有助于确定控制特定区域类固醇产生的潜在新途径。
