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大鼠釉质器官细胞中Na+/K(+)-ATP酶的基因表达谱及定位

Gene-expression profile and localization of Na+/K(+)-ATPase in rat enamel organ cells.

作者信息

Wen Xin, Lacruz Rodrigo S, Smith Charles E, Paine Michael L

机构信息

Center for Craniofacial Molecular Biology, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA, USA.

出版信息

Eur J Oral Sci. 2014 Feb;122(1):21-6. doi: 10.1111/eos.12106. Epub 2013 Dec 7.

Abstract

The sodium pump Na(+)/K(+)-ATPase, expressed in virtually all cells of higher organisms, is involved in establishing a resting membrane potential and in creating a sodium gradient to facilitate a number of membrane-associated transport activities. Na(+)/K(+)-ATPase is an oligomer of α, β, and γ subunits. Four unique genes encode each of the α and β subunits. In dental enamel cells, the spatiotemporal expression of Na(+)/K(+)-ATPase is poorly characterized. Using the rat incisor as a model, this study provides a comprehensive expression profile of all four α and all four β Na(+)/K(+)-ATPase subunits throughout all stages of amelogenesis. Real-time PCR, western blot analysis, and immunolocalization revealed that α1, β1, and β3 are expressed in the enamel organ and that all three are most highly expressed during late-maturation-stage amelogenesis. Expression of β3 was significantly higher than expression of β1, suggesting that the dominant Na(+)/K(+)-ATPase consists of an α1β3 dimer. Localization of α1, β1, and β3 subunits in ameloblasts was primarily to the cytoplasm and occasionally along the basolateral membranes. Weaker expression was also noted in papillary layer cells during early maturation. Our data support that Na(+)/K(+)-ATPase is functional in maturation-stage ameloblasts.

摘要

钠泵Na(+)/K(+)-ATP酶几乎在所有高等生物的细胞中都有表达,它参与建立静息膜电位,并产生钠梯度以促进多种与膜相关的转运活动。Na(+)/K(+)-ATP酶是α、β和γ亚基的寡聚体。α和β亚基各由四个独特的基因编码。在牙釉质细胞中,Na(+)/K(+)-ATP酶的时空表达特征尚不明确。本研究以大鼠切牙为模型,全面展示了在釉质形成的各个阶段,Na(+)/K(+)-ATP酶所有四个α亚基和四个β亚基的表达情况。实时定量PCR、蛋白质印迹分析和免疫定位显示,α1、β1和β3在釉器中表达,且在成熟后期阶段的釉质形成过程中表达量最高。β3的表达显著高于β1,这表明主要的Na(+)/K(+)-ATP酶由α1β3二聚体组成。α1、β1和β3亚基在成釉细胞中的定位主要在细胞质中,偶尔也沿基底外侧膜分布。在成熟早期,乳头层细胞中也观察到较弱的表达。我们的数据支持Na(+)/K(+)-ATP酶在成熟阶段的成釉细胞中发挥功能。

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