Cellular localization of divalent metal transporter DMT-1 in rat kidney
Cellular localization of divalent metal transporter DMT-1 in rat kidney
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DOI：
10.1152/ajprenal.2001.280.5.f803
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发表时间：
2001-05-01
影响因子：
4.2
通讯作者：
Riccardi, D
中科院分区：
文献类型：
--
作者：
Ferguson, CJ;Wareing, M;Riccardi, D
我们已经证明，肾脏在铁平衡中起着重要作用，并且这种离子的代谢显着重吸收发生在亨利循环和集合管中[Wareing M、Ferguson CJ、Green R、Riccardi D和Smith CP。 J Physiol (伦敦) 524: 581-586, 2000]。为了测试二价金抗体并通过蛋白质印迹分析、免疫荧光和共聚焦显微镜测定大鼠肾脏中的 DMT-1 分布。在主细胞和闰细胞的集合管中发现了最强的 DMT1 特异性（即肽保护）免疫反应性。亨利氏袢的粗升肢和更强烈的远端曲管表现出顶端免疫染色。在整个肾单位中，特别是在 S3 段，观察到相当大的细胞内 DMT-1 免疫反应性。所描述的 DMT-1 蛋白分布与我们之前对肾单位铁重吸收位点的鉴定一致，表明 DMT-1 提供了顶端铁进入远端肾单位而不是近端肾小管的分子机制。不同的系统可以促进基底外侧铁的排出。
We have demonstrated that the kidney plays an important role in iron balance and that metabolically significant reabsorption of this ion occurs in the loop of Henle and the collecting ducts [Wareing M, Ferguson CJ, Green R, Riccardi D, and Smith CP. J Physiol (Lond) 524: 581-586, 2000]. To test the possibility that the divalent metal transporter DMT1 (Gunshin H, Mackenzie B, Berger UV, Gunshin Y, Romero MF, Boron WF, Nussberger S, Gollan JL, and Hediger MA. Nature 388: 482-488, 1997) could represent the apical route for iron entry in the kidney, we raised and affinity-purified an anti-DMT-1 polyclonal antibody and determined DMT-1 distribution in rat kidney by Western analysis, immunofluorescence, and confocal microscopy. The strongest DMT1-specific (i.e., peptide-protectable) immunoreactivity was found in the collecting ducts, in both principal and intercalated cells. Thick ascending limbs of Henle's loop and, more intensely, distal convoluted tubules exhibited apical immunostaining. Considerable intracellular DMT-1 immunoreactivity was seen throughout the nephron, particularly in S3 segments. The described distribution of DMT-1 protein is in agreement with our previous identification of nephron sites of iron reabsorption, suggesting that DMT-1 provides the molecular mechanism for apical iron entry in the distal nephron but not in the proximal tubule. Basolateral iron exit may be facilitated by a different system.