PDF(Pubmed)
Abstract：
The cerebrovascular endothelial cells with distinct characteristics line cerebrovascular blood vessels and are the fundamental structure of the blood-brain barrier, which is important for the development and homeostatic maintenance of the central nervous system. Cre-LoxP system-based spatial gene manipulation in mice is critical for investigating the physiological functions of key factors or signaling pathways in cerebrovascular endothelial cells. However, there is a lack of Cre recombinase mouse lines that specifically target cerebrovascular endothelial cells. Here, using a publicly available single-cell RNAseq database, we screened the solute carrier organic anion transporter family member 1a4 (Slco1a4) as a candidate marker of cerebrovascular endothelial cells. Then, we generated an inducible Cre mouse line in which a CreERT2-T2A-tdTomato cassette was placed after the initiation codon ATG of the Slco1a4 locus. We found that tdTomato, which can indicate the endogenous Slco1a4 expression, was expressed in almost all cerebrovascular endothelial cells but not in any other non-endothelial cell types in the brain, including neurons, astrocytes, oligodendrocytes, pericytes, smooth muscle cells, and microglial cells, as well as in other organs. Consistently, when crossing the ROSA26LSL-EYFP Cre reporter mouse, EYFP also specifically labeled almost all cerebrovascular endothelial cells upon tamoxifen induction. Overall, we generated a new inducible Cre line that specifically targets cerebrovascular endothelial cells.
摘要：
脑血管内皮细胞具有明显的排列脑血管的特征,是血脑屏障的基本结构,这对中枢神经系统的发育和稳态维持很重要。基于Cre-LoxP系统的小鼠空间基因操作对于研究脑血管内皮细胞中关键因素或信号通路的生理功能至关重要。然而,缺乏特异性靶向脑血管内皮细胞的Cre重组酶小鼠系。这里,使用公开可用的单细胞RNAseq数据库，我们筛选出溶质载体有机阴离子转运体家族成员1a4(Slco1a4)作为脑血管内皮细胞的候选标志物。然后，我们产生了诱导型Cre小鼠系，其中将CreERT2-T2A-tdTomato盒置于Slco1a4基因座的起始密码子ATG之后。我们发现tdTomato,这可以表明内源性Slco1a4的表达，在几乎所有脑血管内皮细胞中表达，但在大脑中的任何其他非内皮细胞类型中都不表达，包括神经元，星形胶质细胞,少突胶质细胞,周细胞,平滑肌细胞，和小胶质细胞，以及其他器官。始终如一，穿越ROSA26LSL-EYFPCre记者鼠标时，在他莫昔芬诱导后,EYFP还特异性标记几乎所有的脑血管内皮细胞。总的来说,我们产生了一种特异性靶向脑血管内皮细胞的新型诱导型Cre细胞系.
