School of Health Sciences, Purdue University, West Lafayette, IN 47907, USA.
Exp Biol Med (Maywood). 2014 Feb;239(2):202-12. doi: 10.1177/1535370213509213. Epub 2013 Dec 5.
Zinc (Zn) is an essential element for normal brain function; an abnormal Zn homeostasis in brain and the cerebrospinal fluid (CSF) has been implied in the etiology of Alzheimer's disease (AD). However, the mechanisms that regulate Zn transport in the blood-brain interface remain unknown. This study was designed to investigate Zn transport by the blood-CSF barrier (BCB) in the choroid plexus, with a particular focus on Zn transporter-2 (ZnT2), and to understand if lead (Pb) accumulation in the choroid plexus disturbed the Zn regulatory function in the BCB. Confocal microscopy, quantitative PCR and western blot demonstrated the presence of ZnT2 in the choroidal epithelia; ZnT2 was primarily in cytosol in freshly isolated plexus tissues but more toward the peripheral membrane in established choroidal Z310 cells. Exposure of rats to Pb (single ip injection of 50 mg Pb acetate/kg) for 24 h increased ZnT2 fluorescent signals in plexus tissues by confocal imaging and protein expression by western blot. Similar results were obtained by in vitro experiments using Z310 cells. Further studies using cultured cells and a two-chamber Transwell device showed that Pb treatment significantly reduced the cellular Zn concentration and led to an increased transport of Zn across the BCB, the effect that may be due to the increased ZnT2 by Pb exposure. Taken together, these results indicate that ZnT2 is present in the BCB; Pb exposure increases the ZnT2 expression in choroidal epithelial cells by a yet unknown mechanism and as a result, more Zn ions may be deposited into the intracellular Zn pool, leading to a relative Zn deficiency state in the cytoplasm at the BCB.
锌(Zn)是大脑正常功能所必需的元素;脑和脑脊液(CSF)中的异常 Zn 动态平衡与阿尔茨海默病(AD)的病因有关。然而,调节血脑屏障(BCB)中 Zn 转运的机制尚不清楚。本研究旨在研究脉络丛的血脑屏障(BCB)中 Zn 的转运,特别关注 Zn 转运体-2(ZnT2),并了解脉络丛中 Pb 积累是否会干扰 BCB 中的 Zn 调节功能。共聚焦显微镜、定量 PCR 和 Western blot 显示 ZnT2 存在于脉络丛上皮细胞中;新鲜分离的脉络丛组织中 ZnT2 主要位于细胞质中,但在已建立的脉络丛 Z310 细胞中更靠近外周膜。用 50mg/kg 醋酸铅单次腹腔注射大鼠 24 小时,通过共聚焦成像和 Western blot 增加了脉络丛组织中 ZnT2 的荧光信号。用 Z310 细胞进行的体外实验也得到了类似的结果。进一步的细胞培养和双室 Transwell 装置研究表明,Pb 处理显著降低了细胞内 Zn 浓度,并导致 Zn 穿过 BCB 的转运增加,这种作用可能是由于 Pb 暴露导致 ZnT2 增加所致。总之,这些结果表明 ZnT2 存在于 BCB 中;Pb 暴露通过未知机制增加脉络丛上皮细胞中的 ZnT2 表达,结果可能导致更多的 Zn 离子沉积到细胞内 Zn 池中,导致 BCB 细胞质中相对 Zn 缺乏状态。