Peripheral nerve pericytes originating from the blood-nerve barrier expresses tight junctional molecules and transporters as barrier-forming cells
Peripheral nerve pericytes originating from the blood-nerve barrier expresses tight junctional molecules and transporters as barrier-forming cells
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DOI：
10.1002/jcp.21508
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发表时间：
2008-11-01
影响因子：
5.6
通讯作者：
Kanda, Takashi
中科院分区：
文献类型：
--
作者：
Shimizu, Fumitaka;Sano, Yasuteru;Kanda, Takashi
本研究的目的是建立血-神经屏障(BNB)来源的周围神经周细胞系，并研究其作为屏障形成细胞的独特特性。我们从携带温度敏感型猿猴病毒40大T抗原基因的转基因大鼠中分离出周围神经、脑和肺周细胞。这些细胞株表达多种周细胞标志物，如α-平滑肌肌动蛋白、NG2、骨桥蛋白和结蛋白，而不表达血管内皮细胞标志物，如vWF和PECAM。此外，这些细胞株还表达几种紧密连接的分子，如occludin、claudin-12、ZO-1和ZO-2。特别是，在周围神经和脑周细胞中检测到了occludin的表达，尽管通过Western印迹分析在肺周细胞中没有检测到。免疫细胞化学分析证实，occludin和ZO-1定位于周细胞之间的细胞-细胞边界。脑和周围神经周细胞也表现出明显高于肺周细胞的跨周细胞电阻值和更低的菊糖清除率。我们认为，定位于周细胞间细胞-细胞边界的封闭素可能具有机械稳定BNB微血管和血脑屏障的作用。此外，我们还发现这些细胞系表达许多与屏障相关的转运蛋白。用Western印迹法检测ABCG2、p-gp、MRP-1和GLUT-1的表达，免疫细胞化学方法观察细胞胞浆和外膜的表达。周细胞上的这些转运体可能与内皮细胞上的转运体协同促进周围神经到血液的外流和血液到周围神经的内流，以维持周围神经的动态平衡。
The objective of this study was to establish pure blood-nerve barrier (BNB)-derived peripheral nerve pericyte cell lines and to investigate their unique properties as barrier-forming cells. We isolated peripheral nerve, brain, and lung pericytes from transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene. These cell lines expressed several pericyte markers such as alpha-smooth muscle actin, NG2, osteopontin, and desmin, whereas they did not express endothelial cell markers such as vWF and PECAM. In addition, these cell lines expressed several tight junction molecules such as occludin, claudin-12, ZO-1, and ZO-2. In Particular, the expression of occludin was detected in peripheral nerve and brain pericytes,although it was not detected in lung pericytes by a Western blot analysis. An immunocytochemical analysis confirmed that occludin and ZO-1 were localized at the cell-cell boundaries among the pericytes. Brain and peripheral nerve pericytes also showed significantly higher trans-pericyte electrical resistance values and lower inulin clearances than lung pericytes. We considered that occludin localized at the cell-cell boundaries among die pericytes might mechanically stabilize the microvessels of the BNB and the blood-brain barrier. Furthermore, we also showed that these cell lines expressed many barrier-related transporters. ABCG2, p-gp, MRP-1, and Glut-1 were detected by a Western blot analysis and were observed in the cytoplasm and outer membrane by an immunocytochemical analysis. These transporters on pericytes might facilitate the peripheral nerve-to-blood efflux and blood-to-peripheral nerve influx transport of substrates in cooperation with those on endothelial cells in order to maintain peripheral nerve homeostasis.