Exosomal GAPDH from Proximal Tubule Cells Regulate ENaC Activity.
Exosomal GAPDH from Proximal Tubule Cells Regulate ENaC Activity.
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DOI：
10.1371/journal.pone.0165763
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Alli AA
中科院分区：
文献类型：
--
作者：
Jella KK;Yu L;Yue Q;Friedman D;Duke BJ;Alli AA
外泌体是纳米级的细胞来源的囊泡，含有各种分子，包括核酸、蛋白质和脂质。这些囊泡可以将其货物释放到邻近或远处的细胞中，并介导细胞间通讯和细胞功能。在这里，我们检查了从近端小管 LLC-PK1 细胞分离的外泌体对 mpkCCD 细胞和远端小管 Xenopus 2F3 细胞中上皮钠通道的调节。培养的 mpkCCD 细胞用与绿色荧光团偶联的 CTX 进行染色，以标记细胞膜，并用红色亲脂性染料 PKH26 标记从 LLC-PK1 细胞中新鲜分离的外泌体，以便可视化细胞对外泌体的摄取。单通道膜片钳记录显示，爪蟾 2F3 细胞和新分离的裂开小管中 ENaC 的开放概率因外源应用来自 LLC-PK1 近端小管细胞的外泌体而降低。在源自近端小管 LLC-PK1 细胞的外泌体中鉴定出活性 GAPDH。应用转染 GAPDH 抑制剂庚烯酸的外泌体后，非洲爪蟾 2F3 细胞中 ENaC 活性的影响减弱。此外，我们还发现 GAPDH 和 ENaC 亚基在 mpkCCD 细胞中关联。这些研究探讨了外泌体在 ENaC 活性调节中的潜在作用，并探讨了从近端小管细胞到远端小管和集合管细胞通讯的可能机制。
Exosomes are nanometer-scale, cell-derived vesicles that contain various molecules including nucleic acids, proteins, and lipids. These vesicles can release their cargo into adjacent or distant cells and mediate intercellular communication and cellular function. Here we examined the regulation of epithelial sodium channels in mpkCCD cells and distal tubule Xenopus 2F3 cells by exosomes isolated from proximal tubule LLC-PK1 cells. Cultured mpkCCD cells were stained with CTX coupled to a green fluorophore in order to label the cell membranes and freshly isolated exosomes from LLC-PK1 cells were labeled with the red lipophilic dye PKH26 in order to visualize uptake of exosomes into the cells. Single-channel patch clamp recordings showed the open probability of ENaC in Xenopus 2F3 cells and in freshly isolated split-open tubules decreased in response to exogenous application of exosomes derived from LLC-PK1 proximal tubule cells. Active GAPDH was identified within exosomes derived from proximal tubule LLC-PK1 cells. The effect on ENaC activity in Xenopus 2F3 cells was blunted after application of exosomes transfected with the GAPDH inhibitor heptelidic acid. Also, we show GAPDH and ENaC subunits associate in mpkCCD cells. These studies examine a potential role for exosomes in the regulation of ENaC activity and examine a possible mechanism for communication from proximal tubule cells to distal tubule and collecting duct cells.
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影响因子：
2.9
作者：
GLASER, PE;GROSS, RW
通讯作者：
GROSS, RW
影响因子：
2.7
作者：
KUMAGAI, H;SAKAI, H
通讯作者：
SAKAI, H
影响因子：
4.8
作者：
Passero, Christopher J.;Mueller, Gunhild M.;Kleyman, Thomas R.
通讯作者：
Kleyman, Thomas R.
DOI：
10.1152/ajplung.00435.2004
发表时间：
2005-05-01
影响因子：
4.9
作者：
Caldwell, RA;Boucher, RC;Stutts, MJ
通讯作者：
Stutts, MJ
DOI：
10.1152/ajprenal.00403.2007
发表时间：
2008-01-01
影响因子：
4.2
作者：
Pochynyuk, Oleh;Bugaj, Vladislav;Stockand, James D.
通讯作者：
Stockand, James D.