A novel in vitro co-culture model to examine contact formation between astrocytic processes and cerebral vessels
A novel in vitro co-culture model to examine contact formation between astrocytic processes and cerebral vessels
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一种新颖的体外共培养模型，用于检查星形胶质细胞过程和脑血管之间的接触形成
DOI：
10.1016/j.yexcr.2018.12.006
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发表时间：
2019
影响因子：
3.7
通讯作者：
Rikitake Yoshiyuki
中科院分区：
文献类型：
--
作者：
Kawauchi Shoji;Horibe Sayo;Sasaki Naoto;Hirata Ken-ichi;Rikitake Yoshiyuki
在此，我们建立了一种新的体外培养模型，将携带过程的星形胶质细胞和离体的小鼠大脑微血管共培养。成年小鼠和新生小鼠的星形胶质细胞均与微血管形成接触。然而，仅在新生小鼠微血管中检测到水通道蛋白-4 （AQP4）免疫荧光信号在血管周围足端突与血管接触部位的集中定位。星形细胞突起与微血管之间的接触被保留，而AQP4信号在接触部位的集中定位被丢失，通过敲低糖醛酸异常蛋白或α-syntrophin，反映了AQP4在脑血管周围区域的极化定位。此外，通过体外培养模型，我们发现星形胶质细胞主要将突起延伸到位于微血管腔表面的周细胞，这进一步证明了该模型代表了它们的体内情况。总之，我们已经开发了一种新的体外培养模型，可以再现其体内情况的各个方面，并有助于评估星形胶质细胞和血管之间的接触形成。
Here, we developed a novelin vitroco-culture model, in which process-bearing astrocytes and isolated cerebral microvessels from mice were co-cultured. Astrocytes formed contacts with microvessels from both adult and neonatal mice. However, concentrated localization of the immunofluorescence signal for aquaporin-4 (AQP4) at contact sites between perivascular endfoot processes and blood vessels was only detected with neonatal mouse microvessels. Contact between astrocytic processes and microvessels was retained, whereas concentrated localization of AQP4 signal at contact sites was lost, by knockdown of dystroglycan or α-syntrophin, reflecting polarized localization of AQP4 at perivascular regions in the brain. Further, using ourin vitroco-culture model, we found that astrocytes predominantly extend processes to pericytes located at the abluminal surface of microvessels, providing additional evidence that this model is representative of thein vivosituation. Altogether, we have developed a novelin vitroco-culture model that can reproduce aspects of thein vivosituation and is useful for assessing contact formation between astrocytes and blood vessels.