Transcriptomic analysis in cerebral microvessels from the brain of endothelial-specific Crif1 deficiency mouse

Endothelial cells (ECs) in cerebral vessels are considered the primary targets in acute hemorrhagic brain injuries. EC dysfunction can aggravate neuronal injuries by causing secondary inflammatory responses and blood-brain barrier (BBB) disruption. ECs comprising the BBB are known to have a higher mitochondrial volume compared with peripheral ECs. In previous study, we reported Tek-CRIF1-knockout (KO) mice, with EC-specific deletion of the mitochondrial OxPhos-related gene, Crif1, also known as Gadd45gip1 (encoding GADD45G-interacting protein 1), display profound BBB defects accompanied by reduced expression of junctional proteins in ECs. To identify signaling pathways involved in linking EC-specific mitochondrial dysfunction and BBB disruption, we first performed RNA sequencing using isolated cerebral vessels from Tek-CRIF1 mice. This transcriptome analyses of the Tek-CRIF1-KO mouse revealed significant changes in some signaling, a pathway intimately involved in BBB maintenance. RNA was isolated from cerebral microvessels from the brain of Tek- CRIF1-wild type mice (WT) and endothelial-specific Crif1 deletion mice, Tek-CRIF1-knockout mice (KO). (n=3 each per group)脑血管内皮细胞（Endothelial cells, ECs）被视为急性出血性脑损伤的主要靶标。内皮细胞功能异常可通过诱发继发性炎症反应与血脑屏障（blood-brain barrier, BBB）破坏，加重神经元损伤。构成血脑屏障的内皮细胞，其线粒体体积较外周内皮细胞更高，这一特征已被广泛认知。 在既往研究中，我们曾报道Tek-CRIF1基因敲除（knockout, KO）小鼠：该模型通过内皮细胞特异性敲除线粒体氧化磷酸化相关基因Crif1（亦称为Gadd45gip1，编码GADD45G相互作用蛋白1），可引发严重的血脑屏障缺陷，并伴随内皮细胞连接蛋白表达水平下调。 为明确介导内皮细胞特异性线粒体功能异常与血脑屏障破坏之间关联的信号通路，我们首先利用从Tek-CRIF1小鼠中分离的脑血管开展RNA测序（RNA sequencing）。对Tek-CRIF1-KO小鼠的转录组分析显示，多条信号通路发生显著变化，其中一条与血脑屏障维持密切相关的通路尤为突出。 我们从Tek-CRIF1野生型（wild type, WT）小鼠及内皮细胞特异性Crif1敲除的Tek-CRIF1基因敲除（KO）小鼠的脑组织中分离脑微血管并提取RNA，每组各3只小鼠。