Original data of SNHG12-occludin project

We sought to characterize BBB integrity in Snhg12 eKO mice by measuring the leakage of exogenous FITC-dextran tracers. FITC-conjugated 40-kDa or 70-kDa dextran tracer (250μg/g) dissolved in saline was injected to 3-month-old Snhg12 eKO and control mice through the tail vein. Brains and retinas were harvested after 30-min or 40-min circulation. Vascular leakage of brain and retinas to 40-kDa or 70-kDa dextran tracer was quantified by leakage intensity of extravascular fluorescein. The results showed that the brain of 3-month-old Snhg12 eKO mice displayed obvious extravascular accumulation of 40kDa-dextran, whereas the fluorescein was confined to microvessels in control littermates. Meanwhile, we checked the integrity of blood-retina barrier (BRB), an analog of BBB. In the eyes of Snhg12 eKO mice, severe vascular leakage was observed through 40kDa-dextran, but not in the eyes of control mice. The integrity of BBB and BRB were further quantified by measuring the leakage of a larger molecule tracer (70kDa-dextran) in Snhg12 eKO mice. The fluorescent tracer was constrained to the vessels of both brain and retina. .

本研究旨在通过测量外源性FITC-右旋糖酐示踪剂的渗漏来表征Snhg12 eKO小鼠的脑部血脑屏障（BBB）的完整性。将溶解于生理盐水中的FITC偶联的40-kDa或70-kDa右旋糖酐示踪剂（250μg/g）通过尾静脉注射至3个月大的Snhg12 eKO小鼠和对照组小鼠体内。在血液循环30分钟或40分钟后，采集大脑和视网膜。通过测量细胞外荧光素的渗漏强度来量化大脑和视网膜对40-kDa或70-kDa右旋糖酐示踪剂的血管渗漏。结果显示，3个月大的Snhg12 eKO小鼠大脑中40kDa-右旋糖酐的细胞外积累明显，而荧光素在对照组小鼠的微血管中受限。同时，我们检测了血-视网膜屏障（BRB），即BBB的类似物。在Snhg12 eKO小鼠的眼中，通过40kDa-右旋糖酐观察到严重的血管渗漏，而在对照组小鼠的眼中则未观察到。通过测量更大分子量的示踪剂（70kDa-右旋糖酐）在Snhg12 eKO小鼠中的渗漏，进一步量化了BBB和BRB的完整性。荧光示踪剂被限制在脑部和视网膜的血管中。