Department of Surgery, Division of Urology, Cancer Research Program, McGill University Health Centre, 1001 Decarie Boulevard, Montreal, Quebec, H4A 3J1, Canada.
MRC Human Reproductive Sciences Unit, The Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh, EH16 4TJ, UK.
Sci Rep. 2017 Nov 27;7(1):16385. doi: 10.1038/s41598-017-16685-8.
Prostate organogenesis involves epithelial growth controlled by inductive signalling from specialised mesenchymal subsets. To identify pathways active in mesenchyme we used tissue and single cell transcriptomics to define mesenchymal subsets and subset-specific transcript expression. We documented transcript expression using Tag-seq and RNA-seq in female rat Ventral Mesenchymal Pad (VMP) as well as adjacent urethra comprised of smooth muscle and peri-urethral mesenchyme. Transcripts enriched in female VMP were identified with Tag-seq of microdissected tissue, RNA-seq of cell populations, and single cells. We identified 400 transcripts as enriched in the VMP using bio-informatic comparisons of Tag-seq and RNA-seq data, and 44 were confirmed by single cell RNA-seq. Cell subset analysis showed that VMP and adjacent mesenchyme were composed of distinct cell types and that each tissue contained two subgroups. Markers for these subgroups were highly subset specific. Thirteen transcripts were validated by qPCR to confirm cell specific expression in microdissected tissues, as well as expression in neonatal prostate. Immunohistochemical staining demonstrated that Ebf3 and Meis2 showed a restricted expression pattern in female VMP and prostate mesenchyme. We conclude that prostate inductive mesenchyme shows limited cellular heterogeneity and that transcriptomic analysis identified new mesenchymal subset transcripts associated with prostate organogenesis.
前列腺器官发生涉及上皮细胞的生长，受诱导信号的控制，这些信号来自专门的间充质亚群。为了鉴定间充质中活跃的途径，我们使用组织和单细胞转录组学来定义间充质亚群和亚群特异性转录表达。我们使用 Tag-seq 和 RNA-seq 在雌性大鼠腹侧间充质垫（VMP）以及由平滑肌和尿道周围间充质组成的相邻尿道中记录转录物表达。使用 Tag-seq 对微切割组织、细胞群体和单细胞的 RNA-seq 进行了转录物的富集鉴定。我们使用 Tag-seq 和 RNA-seq 数据的生物信息学比较鉴定了 400 个在 VMP 中富集的转录物，其中 44 个通过单细胞 RNA-seq 得到证实。细胞亚群分析表明，VMP 和相邻间充质由不同的细胞类型组成，并且每个组织都包含两个亚群。这些亚群的标记具有高度的亚群特异性。通过 qPCR 验证了 13 个转录本，以确认微切割组织中的细胞特异性表达，以及在新生前列腺中的表达。免疫组织化学染色表明，Ebf3 和 Meis2 在雌性 VMP 和前列腺间充质中表现出受限的表达模式。我们得出结论，前列腺诱导性间充质显示出有限的细胞异质性，转录组分析鉴定了与前列腺器官发生相关的新的间充质亚群转录本。
