Expression of zonula occludens-1 (ZO-1) and the transcription factor ZO-1-associated nucleic acid-binding protein (ZONAB)-MsY3 in glial cells and colocalization at oligodendrocyte and astrocyte gap junctions in mouse brain
Expression of zonula occludens-1 (ZO-1) and the transcription factor ZO-1-associated nucleic acid-binding protein (ZONAB)-MsY3 in glial cells and colocalization at oligodendrocyte and astrocyte gap junctions in mouse brain
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DOI：
10.1111/j.1460-9568.2005.04225.x
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发表时间：
2005-07-01
影响因子：
3.4
通讯作者：
Nagy, JI
中科院分区：
文献类型：
--
作者：
Penes, MC;Li, XB;Nagy, JI
含有PDZ结构域的蛋白质闭合小带-1（ZO-1）与间隙连接形成蛋白的连接蛋白（Cx）家族的几个成员相互作用，并已定位于间隙连接，包括少突胶质细胞中含有Cx 47的间隙连接。我们现在提供的证据ZO-1表达在星形胶质细胞在体内和协会与星形胶质细胞连接蛋白的共聚焦免疫荧光演示ZO-1共定位与星形胶质细胞Cx 30和Cx43，并通过ZO-1免疫共沉淀与Cx 30和Cx43。Cx 30与ZO-1直接相互作用的证据是通过下拉试验获得的，该试验表明Cx 30与ZO-1中三个PDZ结构域中的第二个结合。此外，我们研究了小鼠Y-盒转录因子MsY 3，犬直系同源物已被称为ZO-1相关的核酸结合蛋白（ZONAB），以前报道与ZO-1相互作用。通过使用特异性抗小鼠ZONAB抗体的免疫荧光，发现ZONAB与整个小鼠脑和脊髓中的少突胶质细胞相关，并且与少突胶质细胞Cx47和Cx 32以及星形胶质细胞Cx43共定位。我们的研究结果扩展了表达多功能蛋白ZO-1的CNS细胞类型，证明了一种额外的连接蛋白（Cx 30）与ZO-1直接相互作用，并首次显示了转录因子（ZONAB）与ZO-1定位于少突胶质细胞和星形胶质细胞间隙连接的关联。鉴于以前的观察，ZONAB和ZO-1的组合调节基因表达，我们的研究结果表明，神经胶质细胞间隙连接介导的锚定信号分子在各种各样的神经胶质细胞稳态过程中的作用。
The PDZ domain-containing protein zonula occludens-1 (ZO-1) interacts with several members of the connexin (Cx) family of gap junction-forming proteins and has been localized to gap junctions, including those containing Cx47 in oligodendrocytes. We now provide evidence for ZO-1 expression in astrocytes in vivo and association with astrocytic connexins by confocal immunofluorescence demonstration of ZO-1 colocalization with astrocytic Cx30 and Cx43, and by ZO-1 coimmunoprecipitation with Cx30 and Cx43. Evidence for direct interaction of Cx30 with ZO-1 was obtained by pull-down assays that indicated binding of Cx30 to the second of the three PDZ domains in ZO-1. Further, we investigated mouse Y-box transcription factor MsY3, the canine ortholog of which has been termed ZO-1-associated nucleic acid-binding protein (ZONAB) and previously reported to interact with ZO-1. By immunofluorescence using specific antimouse ZONAB antibody, ZONAB was found to be associated with oligodendrocytes throughout mouse brain and spinal cord, and to be colocalized with oligodendrocytic Cx47 and Cx32 as well as with astrocytic Cx43. Our results extend the CNS cell types that express the multifunctional protein ZO-1, demonstrate an additional connexin (Cx30) that directly interacts with ZO-1, and show for the first time the association of a transcription factor (ZONAB) with ZO-1 localized to oligodendrocyte and astrocyte gap junctions. Given previous observations that ZONAB and ZO-1 in combination regulate gene expression, our results suggest roles of glial gap junction-mediated anchoring of signalling molecules in a wide variety of glial homeostatic processes.