Single-Cell RNA Sequencing Reveals Novel Markers of Male Pituitary Stem Cells and Hormone-Producing Cell Types
Single-Cell RNA Sequencing Reveals Novel Markers of Male Pituitary Stem Cells and Hormone-Producing Cell Types
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DOI：
10.1210/en.2018-00750
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发表时间：
2018-12-01
期刊：
影响因子：
4.8
通讯作者：
Camper, Sally A.
中科院分区：
文献类型：
--
作者：
Cheung, Leonard Y. M.;George, Akima S.;Camper, Sally A.
调节垂体干细胞和特化垂体细胞的转录因子和信号通路一直是深入研究的主题，并对垂体器官发生和疾病产生了机制性的理解。然而，干细胞增殖和分化的调节，特化的前列腺生成细胞之间的异质性，以及非内分泌细胞在腺体中的作用仍然是重要的，未回答的问题。单细胞RNA测序（scRNAseq）技术的最新进展为解决这些问题提供了新的途径。我们对从7周龄C57 BL/6雄性小鼠的六个完整垂体中汇集的13，663个细胞进行scRNAseq。我们在计算机上鉴定了垂体内分泌细胞和干细胞，以及其他支持细胞类型，如内皮细胞、结缔组织、红细胞和白色血细胞。差异基因表达分析确定已知的和新的标记垂体内分泌和干细胞群体。我们通过体内验证一种新的富含促性腺激素的标记物Foxp 2来证明scRNAseq的价值。我们提出了体内垂体组织的新scRNAseq数据，包括来自不可知聚类算法的数据，这些数据表明存在富含固醇/胆固醇合成基因的生长激素亚群。此外，我们表明，不完整的转录组注释可能会导致一些scRNAseq平台上的假阴性，只产生3'转录本末端序列，我们使用体内数据来恢复垂体转录因子Prop 1的读数。最终，scRNAseq技术代表了一个重要的机会，可以解决长期存在的关于脑垂体不同人群一生中的发育和功能的问题。
Transcription factors and signaling pathways that regulate stem cells and specialized hormone-producing cells in the pituitary gland have been the subject of intense study and have yielded a mechanistic understanding of pituitary organogenesis and disease. However, the regulation of stem cell proliferation and differentiation, the heterogeneity among specialized hormone-producing cells, and the role of nonendocrine cells in the gland remain important, unanswered questions. Recent advances in single-cell RNA sequencing (scRNAseq) technologies provide new avenues to address these questions. We performed scRNAseq on; 13,663 cells pooled from six whole pituitary glands of 7-week-old C57BL/6 male mice. We identified pituitary endocrine and stem cells in silico, as well as other support cell types such as endothelia, connective tissue, and red and white blood cells. Differential gene expression analyses identify known and novel markers of pituitary endocrine and stem cell populations. We demonstrate the value of scRNAseq by in vivo validation of a novel gonadotrope-enriched marker, Foxp2. We present novel scRNAseq data of in vivo pituitary tissue, including data from agnostic clustering algorithms that suggest the presence of a somatotrope subpopulation enriched in sterol/cholesterol synthesis genes. Additionally, we show that incomplete transcriptome annotation can cause false negatives on some scRNAseq platforms that only generate 3' transcript end sequences, and we use in vivo data to recover reads of the pituitary transcription factor Prop1. Ultimately, scRNAseq technologies represent a significant opportunity to address long-standing questions regarding the development and function of the different populations of the pituitary gland throughout life.