Quantitative and qualitative assessment of glymphatic flux using Evans blue albumin.
Quantitative and qualitative assessment of glymphatic flux using Evans blue albumin.
复制标题
DOI：
10.1016/j.jneumeth.2018.09.031
复制
发表时间：
2019-01-01
影响因子：
3
通讯作者：
Clark RSB
中科院分区：
文献类型：
--
作者：
Wolf MS;Chen Y;Simon DW;Alexander H;Ross M;Gibson GA;Manole MD;Bayır H;Kochanek PM;Clark RSB
胶质淋巴系统是从脑中清除蛋白质和大分子的建议途径，并且破坏的胶质淋巴流与神经系统疾病有关。我们利用伊文思蓝的比色、荧光和蛋白结合特性来评估胶质淋巴流量。将25 μL溶于人工脑脊液（aCSF）的1%伊文思蓝标记白蛋白（EBA）注入出生后17天麻醉大鼠的脑池内。在注射后的不同时间点收集血清（n=37），并通过荧光定量法测量EBA。在单独的大鼠（n = 3）中，在顶叶皮质上放置颅窗，并使用体内多光子显微镜评价EBA转运。对单独的大鼠（n = 6）进行免疫组织化学处理以检查EBA的定位。在一些大鼠中，通过脑池内注射aCSF增加颅内压（ICP）。EBA检测血清早在30分钟，是最大的在4小时，并在72小时后检测不到脑池内注射。使用活体显微镜和免疫组织化学EBA可以跟踪从CSF到血管周围的位置。与通过胶质淋巴流清除一致，将ICP增加至40 mmHg加速了EBA从CSF到血液的转运。EBA从CSF到血清的转运可以在没有放射性标记的情况下通过荧光定量法进行定量。还可以使用EBA荧光定性地评估淋巴通量。我们提出了一种新的技术，同时定量和定性评价胶质淋巴流量的大鼠。
The glymphatic system is a proposed pathway for clearance of proteins and macromolecules from brain, and disrupted glymphatic flux is implicated in neurological disease. We capitalized on colorimetric, fluorescent, and protein-binding properties of Evans blue to evaluate glymphatic flux. Twenty-five μL of 1% Evans blue-labeled albumin (EBA) in artificial cerebrospinal fluid (aCSF) was injected into the intracisternal space of anesthetized postnatal day 17 rats. Serum was collected at various time points after injection (n=37) and EBA was measured spectrophotometrically. In separate rats (n = 3), a cranial window was placed over the parietal cortex and EBA transit was evaluated using in vivo multiphoton microscopy. Separate rats (n = 6) were processed for immunohistochemistry to examine localization of EBA. In some rats, intracranial pressure (ICP) was increased via intracisternal injection of aCSF. EBA was detected in serum as early as 30 min, was maximal at 4 h, and was undetectable at 72 h after intracisternal injection. Using intra-vital microscopy and immunohistochemistry EBA could be tracked from CSF to perivascular locations. Consistent with removal via glymphatic flux, increasing ICP to 40 mmHg accelerated transit of EBA from CSF to blood. Transit of EBA from CSF to serum could be quantified spectrophotometrically without radioactive labeling. Glymphatic flux could also be qualitatively evaluated using EBA fluorescence. We present a novel technique for simultaneous quantitative and qualitative evaluation of glymphatic flux in rats.
登录
查看更多内容
影响因子：
3
作者：
SARIA, A;LUNDBERG, JM
通讯作者：
LUNDBERG, JM
影响因子：
--
作者：
SZENTISTVANYI, I;PATLAK, CS;CSERR, HF
通讯作者：
CSERR, HF
影响因子：
6.3
作者：
Jiang, Quan;Zhang, Li;Zhang, Zhenggang
通讯作者：
Zhang, Zhenggang
影响因子：
--
作者：
BRADBURY, MWB;CSERR, HF;WESTROP, RJ
通讯作者：
WESTROP, RJ
影响因子：
6.1
作者：
Peng W;Achariyar TM;Li B;Liao Y;Mestre H;Hitomi E;Regan S;Kasper T;Peng S;Ding F;Benveniste H;Nedergaard M;Deane R
通讯作者：
Deane R