Expression and polarity reversal of V-type H+-ATPase during the mineralization–demineralization cycle in Porcellio scaber sternal epithelial cells
Expression and polarity reversal of V-type H+-ATPase during the mineralization–demineralization cycle in Porcellio scaber sternal epithelial cells
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Porcellio scaber胸骨上皮细胞矿化-脱矿循环过程中V型H+-ATP酶的表达和极性逆转
DOI：
10.1242/jeb.00953
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发表时间：
2004
影响因子：
2.8
通讯作者：
Reiner Bleher
中科院分区：
文献类型：
--
作者：
A. Ziegler;D. Weihrauch;M. Hagedorn;D. Towle;Reiner Bleher
摘要 上皮细胞层形成和吸收 CaCO3 需要质子的上皮运输。我们使用陆生等足动物 Porcellio scaber 的前胸骨上皮作为模型，研究间歇性 CaCO3 沉积物矿化和脱矿过程中 V 型 H+-ATPase 的表达模式和免疫定位。半定量RT-PCR显示，从非Ca2+转运控制阶段到CaCO3沉积形成和吸收阶段，V型H+-ATP酶的表达增加。在 Ca2+ 转运阶段，胸骨前部上皮的表达量大于后部胸骨上皮，胸骨上皮不参与沉积物形成，仅转运中等量的 CaCO3。胸骨前上皮中 V 型 H+-ATPase B 亚基的免疫细胞化学显示，基底外侧膜内蛋白质的丰度增加，分别从控制阶段的不可检测信号到 CaCO3 沉积形成阶段的强信号。从 CaCO3 沉积形成阶段到 CaCO3 再吸收阶段，基底外侧质膜内的信号减弱，顶端质膜内的信号增强。结果首次表明 V 型 H+-ATP 酶对 CaCO3 沉积的贡献以及同一细胞内其极性从基底外侧到顶端质膜室的逆转。
SUMMARY The formation and resorption of CaCO3 by epithelial cell layers require epithelial transport of protons. We used the anterior sternal epithelium of the terrestrial isopod Porcellio scaber as a model to study the expression pattern and immunolocalization of a V-type H+-ATPase during the mineralization and demineralization of intermittent CaCO3 deposits. Semiquantitative RT-PCR revealed that the expression of the V-type H+-ATPase increases from non Ca2+-transporting control stages to the stages of CaCO3 deposit formation and resorption. In the Ca2+-transporting stages the expression was larger in the anterior than in the posterior sternal epithelium, which is not involved in deposit formation and transports just moderate amounts of CaCO3. Immunocytochemistry of the B-subunit of the V-type H+-ATPase in the anterior sternal epithelium reveals an increase in the abundance of the protein within the basolateral membrane, from undetectable to strong signals in the control stage to the stages of CaCO3 deposit formation, respectively. From the stage of CaCO3 deposit formation to that of CaCO3 resorption the signal decreased within the basolateral plasma membrane and increased within the apical plasma membrane. For the first time the results indicate a contribution of a V-type H+-ATPase to CaCO3 deposition and a reversal of its polarity from the basolateral to the apical plasma membrane compartment within the same cells.
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DOI：
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