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首页> 外文期刊>American Journal of Physiology >Immunolocalization of aquaporin-8 in rat kidney, gastrointestinal tract, testis, and airways.
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Immunolocalization of aquaporin-8 in rat kidney, gastrointestinal tract, testis, and airways.

机译：Aquaporin-8在大鼠肾脏，胃肠道，睾丸和气道中的免疫定位。

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The purpose of this study was to determine the cellular and subcellular localization of aquaporin-8 (AQP8) in rat kidney and other organs by RT-PCR analyses and by immunoblotting and immunohistochemistry using peptide-derived rabbit antibodies to rat AQP8. RT-PCR and Southern blotting revealed the presence of AQP8 mRNA in all kidney zones. LLC-PK(1) cells transfected with a rat AQP8 construct exhibited strong labeling with the affinity-purified antibodies, whereas controls using cells transfected with the vector, but without the insert, were negative. The labeling was almost exclusively associated with intracellular vesicles. Immunoblotting of kidney membrane fractions revealed a predominant single band of 26-28 kDa. AQP8 immunoreactivity was mainly present in the cortex and outer stripe of the outer medulla. Sequential ultracentrifugation of rat kidney membrane revealed that AQP8 resides predominantly in intracellular vesicular fractions. Immunocytochemistry revealed modest labeling of proximal tubules and weak labeling of collecting ducts in cortex and medulla of rat kidney. The labeling was confined to cytoplasmic areas with no labeling of the brush border. Immunoblotting and RT-PCR/Southern blotting also revealed the presence of AQP8 protein and mRNA in rat liver, testis, epididymis, duodenum, jejunum, colon, and bronchi/trachea. Consistent with this, immunohistochemistry revealed AQP8 labeling in the hepatocytes and spermatogenic cells in testis and in the basal cells in ductus epididymis, trachea, and bronchial epithelia. Moreover, AQP8 labeling was observed in the myoepithelial cells in salivary, bronchial, and tracheal glands with no labeling of acini or ductal epithelial cells. AQP8 is also present in the surface epithelial cells in duodenum, jejunum, and colon. In conclusion, AQP8 is expressed at low levels in rat kidney proximal tubules and collecting ducts, and it is present in distinct cell types in liver, testis, epididymis, duodenum, jejunum, colon, trachea, and principal bronchi as well as in multiple glands, including salivary glands.
机译：这项研究的目的是通过RT-PCR分析以及使用肽源的兔抗大鼠AQP8抗体的免疫印迹和免疫组织化学测定大鼠肾脏和其他器官中水通道蛋白8（AQP8）的细胞和亚细胞定位。 RT-PCR和Southern印迹显示所有肾脏区域均存在AQP8 mRNA。用大鼠AQP8构建体转染的LLC-PK（1）细胞用亲和纯化的抗体表现出较强的标记，而使用用载体转染但未插入的细胞进行的对照则为阴性。标记几乎完全与细胞内囊泡相关。肾膜部分的免疫印迹显示26-28 kDa的主要单条带。 AQP8免疫反应性主要存在于外延髓的皮质和外条纹中。大鼠肾脏膜的顺序超速离心显示，AQP8主要存在于细胞内水泡部分中。免疫细胞化学显示大鼠肾皮质和延髓中近端小管的标记适中，而集合管的标记较弱。标记仅限于细胞质区域，未标记刷状边界。免疫印迹和RT-PCR / Southern印迹法还显示大鼠肝脏，睾丸，附睾，十二指肠，空肠，结肠和支气管/气管中存在AQP8蛋白和mRNA。与此相一致，免疫组织化学显示睾丸的肝细胞和生精细胞以及附睾导管，气管和支气管上皮的基底细胞中的AQP8标记。此外，在唾液，支气管和气管腺的肌上皮细胞中观察到AQP8标记，而没有腺泡或导管上皮细胞标记。 AQP8也存在于十二指肠，空肠和结肠的表面上皮细胞中。总之，AQP8在大鼠肾脏近端小管和收集管中低水平表达，并存在于肝脏，睾丸，附睾，十二指肠，空肠，结肠，气管和主要支气管以及多个腺体的不同细胞类型中，包括唾液腺。

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