Global transcriptional profiles of abdominal skin dermis during perinatal period. Global transcriptional profiles of abdominal skin dermis during perinatal period

We report global transcriptional alteration and transcriptional heterogeneity of dermal cells in remodeling mouse abdominal skin during perinatal period. By single cell RNA sequencing of whole dermal cells isolated from the abdominal skin dermis of non-pregnant (NP) mice, pregnant mice at dpc16, and post-partum mice at dpp42 by using Col1a2creERT2;R26H2B-EGFP mice. By unsupervised evaluation of clustering-based cell identities of total samples on the Seurat platform, we identified 13 main clusters. The second round of unsupervised clustering of each cluster revealed that the vascular cluster exhibits dynamic transcriptome alteration during pregnancy. Overall design: We treated the mice with tamoxifen during pregnancy and harvested abdominal skin at dpc16 and dpp42. For the control, abdominal skin was harvested from non-pregnant (NP) mice treated with tamoxifen for 3 days and harvested at 48 days after treatment. After dissociation of the dermis into single cells, we generated chromium single cell 3′ v2 libraries from randomly captured dermal cells and sequenced them using Illumina NextSeq500. We profiled 6,020 cells (NP: 1,289cells; Dpc16: 3,650cells; Dpp42: 1,081cells) with a range of 4,500–5,400 mean reads per cell for each sample, whereby approximately 1,200–1,500 median genes per cell for each sample were detected.

本研究报道了围产期小鼠腹部皮肤重塑过程中，真皮细胞的全局转录组改变与转录异质性。本研究使用Col1a2creERT2;R26H2B-EGFP转基因小鼠，分别从未孕（non-pregnant, NP）小鼠、孕16天（dpc16）小鼠以及产后42天（dpp42）小鼠的腹部皮肤真皮中分离全真皮细胞，进行单细胞RNA测序（single cell RNA sequencing）。基于Seurat平台对所有样本的基于聚类的细胞身份进行无监督评估后，我们共鉴定出13个主要细胞簇。对每个细胞簇进行第二轮无监督聚类分析后发现，血管细胞簇在妊娠过程中呈现动态的转录组改变。

实验设计总览：我们在妊娠期间对小鼠施加他莫昔芬处理，并分别在孕16天和产后42天收取腹部皮肤样本。对照组则选取经3天他莫昔芬处理的未孕小鼠，于处理后48天收取腹部皮肤样本。将真皮组织解离为单细胞后，我们从随机捕获的真皮细胞中构建Chromium单细胞3'端v2文库（chromium single cell 3′ v2 libraries），并使用Illumina NextSeq500测序平台进行测序。本研究共分析了6020个细胞（未孕组：1289个；孕16天组：3650个；产后42天组：1081个），每个样本的平均测序读取数为4500~5400，每个细胞的中位检测基因数约为1200~1500。