Alterations of the perivascular dystrophin-dystroglycan complex following brain lesions. An immunohistochemical study in rats
Alterations of the perivascular dystrophin-dystroglycan complex following brain lesions. An immunohistochemical study in rats
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DOI：
10.14670/hh-26.1435
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发表时间：
2011-11-01
影响因子：
2
通讯作者：
Sadeghian, S.
中科院分区：
文献类型：
--
作者：
Kalman, M.;Mahalek, J.;Sadeghian, S.
抗肌营养不良蛋白聚糖是一种层粘连蛋白受体，它与抗肌营养不良蛋白和其他成分形成抗肌营养不良蛋白-抗肌营养不良蛋白聚糖复合体。它在胶质血管连接、脑血管形成和血脑屏障的形成中起着重要作用。营养不良多糖由两个亚单位组成，即α和β。以往的研究表明，脑血管的β-肌营养不良蛋白-葡聚糖免疫反应在邻近病变的区域暂时消失，而在完整的脑组织中没有免疫反应的血管层粘连蛋白变得可检测到。本研究将这些研究扩展到该复合体的其他成分：促性腺激素、α1-合体营养素和α1-营养不良素。实验是在成年大鼠身上进行的。病变为深部氯胺酮-木素麻醉下的刺伤或冷冻灶。存活2~30d后，灌流漂浮脑片进行荧光免疫组织化学染色。α1-dystrobrevin和β-dystroglan一样，在病变周围暂时消失。促性腺激素的免疫反应性变化与层粘连蛋白的变化相似。在完整的大脑中，它们被限制在进入血管的节段和脑室周围器官。皮损后，它们的免疫反应在皮损周围的血管中表现出来。然而，促性腺激素在层粘连蛋白之后有一个延迟：它们的峰值分别在术后第21天和第7天左右。阳性星形胶质细胞中仅有α1-合成素免疫反应阳性，在14天时达到高峰。双标记证实其与GFAP共定位。低温损伤具有相似的免疫组织化学效应，但为Western印迹分析提供了更合适的样本，证实了α1-dystrobrevin和α1-syntroin水平的改变。这些现象可能有助于监测损伤后血管突起和胶质血管连接的变化。
Dystroglycan is a laminin receptor, which with dystrophins and other components forms the dystrophin-dystroglycan complex. It has an important role in the formation of gliovascular connections, cerebral vascularisation and blood-brain barrier. Dystroglycan consists of two sub-units, alpha and beta. Previous studies demonstrated that the beta-dystroglycan immunoreactivity of cerebral vessels temporarily disappeared in the area adjacent to the lesion, whereas the vascular laminin which is not immunoreactive in the intact brain became detectable. The present study extends these investigations over other components of the complex: utrophin, alpha 1-syntrophin and alpha 1-dystrobrevin. The experiments were performed on adult rats. The lesions were stab wounds or cryogenic lesions in deep ketamine-xylasine narcosis. Following survival periods 2 to 30 days, the animals were perfused and floating brain sections were processed for fluorescent immunohistochemistry. The alpha 1-dystrobrevin, like beta-dystroglycan, vanished temporarily around the lesion. The immunoreactivity of utrophin changed in a similar way to that of laminin. In intact brains they were confined to the entering segments of the vessels and to the circumventricular organs. Following lesions their immunoreactivity manifested in the vessels around the lesions. However, utrophin followed laminin with a delay: their peaks were about POD (postoperative days) 21 and 7, respectively. Only immunoreactivity of alpha 1-syntrophin appeared in the reactive astrocytes, peaking at POD 14. Double-labeling proved its co-localization with GFAP. Cryogenic lesions had similar immunohistochemical effects, but provided more suitable samples for Western blot analysis, which proved the altered levels of alpha 1-dystrobrevin and alpha 1-syntrophin. The phenomena may help to monitor the post-lesion vascular processes and the alterations of the gliovascular connections.