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III 型钠依赖性磷酸盐转运体 2 在小鼠脑内的定位。

Localization of type-III sodium-dependent phosphate transporter 2 in the mouse brain.

机构信息

Laboratory of Medical Therapeutics and Molecular Therapeutics, Gifu Pharmaceutical University, Gifu, Japan.

出版信息

Brain Res. 2013 Sep 19;1531:75-83. doi: 10.1016/j.brainres.2013.07.038. Epub 2013 Jul 30.

Abstract

Type-III sodium-dependent phosphate transporters 1 and 2 (PiT-1 and PiT-2, respectively) are proteins encoded by SLC20A1 and SLC20A2, respectively. The ubiquitous distribution of PiT-1 and PiT-2 mRNAs in mammalian tissues is in agreement with the housekeeping maintenance of homeostasis of intracellular inorganic phosphate (Pi), which is absorbed from interstitial fluid for normal cellular functions. Recently, mutations of SLC20A2 have been found in patients with idiopathic basal ganglia calcification (IBGC), also known as Fahr's disease. However, the localization of PiT-2 in the brain has not been clarified yet. Therefore, the aim of this study is to clarify the distribution of PiT-2 expression in the mouse brain. Our biochemical and immunohistochemical analyses using a polyclonal antibody (Ab) and a monoclonal Ab showed that PiT-2 was ubiquitously expressed throughout the brain. In terms of the cellular type, PiT-2 was predominantly detected in neurons; it colocalized with β-tubulin III in the cerebral cortex and with calbindin D-28K in Purkinje cells. Additionally, PiT-2 immunopositivity was detected in the microtubule-associated protein 2-positive neuronal dendrites in the cerebral cortex. However, colocalization with PiT-2 immunopositivity was not observed in the synaptophysin-positive nerve terminals. PiT-2 was also expressed in astrocytes and vascular endothelial cells. Our results indicate that PiT-2 plays an important role in the maintenance of cellular Pi homeostasis in neurons, astrocytes, and endothelial cells. This finding is a milestone in the study of the function of PiT-2 in the normal mouse brain and particularly in the brains of patients with Fahr's disease.

摘要

III 型钠依赖磷酸盐转运蛋白 1 和 2(分别为 PiT-1 和 PiT-2)是由 SLC20A1 和 SLC20A2 分别编码的蛋白质。PiT-1 和 PiT-2mRNA 在哺乳动物组织中的广泛分布与细胞内无机磷酸盐(Pi)的维持一致,Pi 从间质液中吸收以维持正常细胞功能。最近,SLC20A2 的突变已在特发性基底节钙化(IBGC)患者中发现,也称为 Fahr 病。然而,PiT-2 在大脑中的定位尚未阐明。因此,本研究旨在阐明 PiT-2 在小鼠大脑中的表达分布。我们使用多克隆抗体(Ab)和单克隆 Ab 进行的生化和免疫组织化学分析表明,PiT-2 在整个大脑中广泛表达。就细胞类型而言,PiT-2 主要在神经元中检测到;它与大脑皮质中的β-微管蛋白 III 以及浦肯野细胞中的钙结合蛋白 D-28K 共定位。此外,在大脑皮质中微管相关蛋白 2 阳性神经元树突中检测到 PiT-2 免疫阳性。然而,在突触小体蛋白阳性神经末梢中未观察到与 PiT-2 免疫阳性的共定位。PiT-2 也在星形胶质细胞和血管内皮细胞中表达。我们的结果表明,PiT-2 在神经元、星形胶质细胞和内皮细胞中维持细胞 Pi 内稳态方面发挥着重要作用。这一发现是研究 PiT-2 在正常小鼠大脑中的功能,特别是在 Fahr 病患者大脑中的功能的一个里程碑。

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