Department of Life Science and Medical Bioscience, Waseda University, Tokyo, Japan.
PLoS One. 2013 Sep 27;8(9):e73685. doi: 10.1371/journal.pone.0073685. eCollection 2013.
Endothelial cells (ECs) lining the blood vessels serve a variety of functions and play a central role in the homeostasis of the circulatory system. Since the ductus arteriosus (DA) has different arterial characteristics from its connecting vessels, we hypothesized that ECs of the DA exhibited a unique gene profile involved in the regulation of DA-specific morphology and function. Using a fluorescence-activated cell sorter, we isolated ECs from pooled tissues from the DA or the descending aorta of Wistar rat fetuses at full-term of gestation (F group) or neonates 30 minutes after birth (N group). Using anti-CD31 and anti-CD45 antibodies as cell surface markers for ECs and hematopoietic derived cells, respectively, cDNAs from the CD31-positive and CD45-negative cells were hybridized to the Affymetrix GeneChip® Rat Gene 1.0 ST Array. Among 26,469 gene-level probe sets, 82 genes in the F group and 81 genes in the N group were expressed at higher levels in DA ECs than in aortic ECs (p<0.05, fold change>2.0). In addition to well-known endothelium-enriched genes such as Tgfb2 and Vegfa, novel DA endothelium-dominant genes including Slc38a1, Capn6, and Lrat were discovered. Enrichment analysis using GeneGo MetaCore software showed that DA endothelium-related biological processes were involved in morphogenesis and development. We identified many overlapping genes in each process including neural crest-related genes (Hoxa1, Hoxa4, and Hand2, etc) and the second heart field-related genes (Tbx1, Isl1, and Fgf10, etc). Moreover, we found that regulation of epithelial-to-mesenchymal transition, cell adhesion, and retinol metabolism are the active pathways involved in the network via potential interactions with many of the identified genes to form DA-specific endothelia. In conclusion, the present study uncovered several significant differences of the transcriptional profile between the DA and aortic ECs. Newly identified DA endothelium-dominant genes may play an important role in DA-specific functional and morphologic characteristics.
血管内皮细胞（ECs）排列在血管内，具有多种功能，在循环系统的稳态中起着核心作用。由于动脉导管（DA）具有与连接血管不同的动脉特征，我们假设 DA 的 ECs 表现出独特的基因谱，参与调节 DA 特异性形态和功能。我们使用荧光激活细胞分选，从 Wistar 胎鼠足月（F 组）或出生后 30 分钟（N 组）的 DA 或降主动脉的汇集组织中分离 ECs。使用抗 CD31 和抗 CD45 抗体作为 ECs 和造血衍生细胞的细胞表面标志物，分别从 CD31 阳性和 CD45 阴性细胞中杂交 cDNA 到 Affymetrix GeneChip®Rat Gene 1.0 ST 阵列。在 26469 个基因水平探针集中，F 组 82 个基因和 N 组 81 个基因在 DA ECs 中的表达水平高于主动脉 ECs（p<0.05，倍数变化>2.0）。除了众所周知的富含内皮细胞的基因，如 Tgfb2 和 Vegfa 外，还发现了新的 DA 内皮细胞优势基因，包括 Slc38a1、Capn6 和 Lrat。使用 GeneGo MetaCore 软件进行的富集分析表明，DA 内皮细胞相关的生物学过程涉及形态发生和发育。我们在每个过程中都发现了许多重叠的基因，包括神经嵴相关基因（Hoxa1、Hoxa4 和 Hand2 等）和第二心脏场相关基因（Tbx1、Isl1 和 Fgf10 等）。此外，我们发现上皮-间质转化、细胞粘附和视黄醇代谢的调节是通过与许多鉴定基因的潜在相互作用形成 DA 特异性内皮的网络中的活跃途径。总之，本研究揭示了 DA 和主动脉 ECs 之间转录谱的几个显著差异。新鉴定的 DA 内皮细胞优势基因可能在 DA 特异性功能和形态特征中发挥重要作用。
