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首页> 外文期刊>The European Journal of Neuroscience >Effects of agrin on the expression and distribution of the water channel protein aquaporin-4 and volume regulation in cultured astrocytes.
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Effects of agrin on the expression and distribution of the water channel protein aquaporin-4 and volume regulation in cultured astrocytes.

机译：凝集素对培养的星形胶质细胞中水通道蛋白aquaporin-4的表达和分布以及体积调节的影响。

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Agrin is a heparan sulfate proteoglycan of the extracellular matrix and is known for organizing the postsynaptic differentiation of the neuromuscular junction. Increasing evidence also suggests roles for agrin in the developing CNS, including the formation and maintenance of the blood-brain barrier. Here we describe effects of agrin on the expression and distribution of the water channel protein aquaporin-4 (AQP4) and on the swelling capacity of cultured astrocytes of newborn mice. If astrocytes were cultured on a substrate containing poly DL-ornithine, anti-AQP4 immunoreactivity was evenly and diffusely distributed. If, however, astrocytes were cultured in the presence of agrin-conditioned medium, we observed an increase in the intensity of AQP4-specific membrane-associated staining. Freeze-fracture studies revealed a clustering of orthogonal arrays of particles, representing a structural equivalent of AQP4, when exogenous agrin was present in the astrocyte cultures. Neuronal and non-neuronal agrin isoforms (agrin A0B0 and agrin A4B8, respectively) were able to induce membrane-associated AQP4 staining. Water transport capacity as well as the density of orthogonal arrays of intramembranous particles was increased in astrocytes cultured with the neuronal agrin isoform A4B8, but not with the endothelial and meningeal isoform A0B0. RT-PCR demonstrated that agrin A4B8 increased the level of the M23 splice variant of AQP4 and decreased the level of the M1 splice variant of AQP4. Implications for the regulation and maintenance of the blood-brain barrier including oedema formation under pathological conditions are discussed.
机译：Agrin是细胞外基质的硫酸乙酰肝素蛋白聚糖，已知用于组织神经肌肉接头的突触后分化。越来越多的证据还表明，凝集素在发展中的中枢神经系统中的作用，包括血脑屏障的形成和维持。在这里，我们描述了凝集素对水通道蛋白aquaporin-4（AQP4）的表达和分布以及新生小鼠培养的星形胶质细胞膨胀能力的影响。如果将星形胶质细胞培养在含有聚DL-鸟氨酸的底物上，则抗AQP4免疫反应性会均匀分散地分布。但是，如果在存在凝集素的培养基中培养星形胶质细胞，我们会观察到AQP4特异性膜相关染色的强度增加。冷冻断裂研究显示，当星形胶质细胞培养物中存在外源凝集素时，颗粒的正交阵列聚集在一起，代表了AQP4的结构等同物。神经元和非神经元凝集素亚型（分别为凝集素A0B0和凝集素A4B8）能够诱导膜相关的AQP4染色。在神经元凝集素同种型A4B8培养的星形胶质细胞中，水运输能力以及膜内颗粒正交阵列的密度增加，但在内皮和脑膜同种型A0B0中却没有。 RT-PCR表明，凝集素A4B8增加了AQP4的M23剪接变体的水平，并降低了AQP4的M1剪接变体的水平。讨论了在病理条件下调节和维持血脑屏障包括水肿形成的意义。

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