Regulation of multidrug resistance-associated protein 2 by calcium signaling in mouse liver.
Regulation of multidrug resistance-associated protein 2 by calcium signaling in mouse liver.
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DOI：
10.1002/hep.23625
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发表时间：
2010-07
期刊：
影响因子：
13.5
通讯作者：
Nathanson, Michael H.
中科院分区：
文献类型：
--
作者：
Cruz, Laura N.;Guerra, Mateus T.;Kruglov, Emma;Mennone, Albert;Garcia, Celia R. S.;Chen, Ju;Nathanson, Michael H.
多药耐药相关蛋白 2 (Mrp2) 是一种负责将有机阴离子分泌到胆汁中的小管转运蛋白。 Mrp2 活性通过插入质膜来调节；然而，控制这一现象的因素尚不清楚。钙 (Ca2+) 信号传导调节大多数细胞类型中囊泡的胞吐作用，II 型肌醇 1,4,5-三磷酸受体 (InsP3R2) 调节肝细胞小管区域的 Ca2+ 释放。然而，InsP3R2 和 Ca2+ 信号在小管插入和 Mrp2 功能中的作用尚不清楚。本研究的目的是确定 InsP3R2 介导的 Ca2+ 信号在将 Mrp2 靶向泪管膜中的作用。使用野生型 (WT) 和 InsP3R2 敲除 (KO) 小鼠的肝脏、分离的肝细胞和胶原夹心培养物中的肝细胞进行蛋白质印迹、共聚焦免疫荧光以及 Ca2+ 信号和荧光有机阴离子分泌的延时成像。通过全内反射显微镜监测HepG2细胞中表达的绿色荧光蛋白(GFP)-Mrp2的质膜插入。 InsP3R2集中在WT小鼠的小管区域，但在InsP3R2 KO肝脏中不存在，而InsP3R1的表达和定位得以保留，并且InsP3R3在WT和KO肝脏中均不存在。在缺乏 InsP3R2 的肝细胞中，三磷酸腺苷 (ATP) 或加压素诱导的 Ca2+ 信号受损。在 KO 肝细胞以及用 1,2-双（邻氨基苯氧基）乙烷-N,N,N',N'-四乙酸 (BAPTA) 处理的 WT 肝细胞中，有机阴离子 5-氯甲基荧光素二乙酸酯 (CMFDA) 的小管分泌减少。此外，InsP3R2 KO 小鼠中牛磺熊去氧胆酸 (TUDCA) 的利胆作用受损。最后，ATP 增加了 HepG2 细胞质膜中的 GFP-Mrp2 荧光，而 BAPTA 也减少了这种荧光。 InsP3R2 介导的 Ca2+ 信号通过将 Mrp2 靶向胆管膜来增强有机阴离子分泌到胆汁中。
Multidrug resistance associated protein 2 (Mrp2) is a canalicular transporter responsible for organic anion secretion into bile. Mrp2 activity is regulated by insertion into the plasma membrane; however, the factors that control this are not understood. Calcium (Ca2+) signaling regulates exocytosis of vesicles in most cell types, and the type II inositol 1,4,5-triphosphate receptor (InsP3R2) regulates Ca2+ release in the canalicular region of hepatocytes. However, the role of InsP3R2 and of Ca2+ signals in canalicular insertion and function of Mrp2 is not known. The aim of this study was to determine the role of InsP3R2-mediated Ca2+ signals in targeting Mrp2 to the canalicular membrane. Livers, isolated hepatocytes, and hepatocytes in collagen sandwich culture from wild-type (WT) and InsP3R2 knockout (KO) mice were used for western blots, confocal immunofluorescence, and time-lapse imaging of Ca2+ signals and of secretion of a fluorescent organic anion. Plasma membrane insertion of green fluorescent protein (GFP)-Mrp2 expressed in HepG2 cells was monitored by total internal reflection microscopy. InsP3R2 was concentrated in the canalicular region of WT mice but absent in InsP3R2 KO livers, whereas expression and localization of InsP3R1 was preserved, and InsP3R3 was absent from both WT and KO livers. Ca2+ signals induced by either adenosine triphosphate (ATP) or vasopressin were impaired in hepatocytes lacking InsP3R2. Canalicular secretion of the organic anion 5-chloromethylfluorescein diacetate (CMFDA) was reduced in KO hepatocytes, as well as in WT hepatocytes treated with 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA). Moreover, the choleretic effect of tauroursodeoxycholic acid (TUDCA) was impaired in InsP3R2 KO mice. Finally, ATP increased GFP-Mrp2 fluorescence in the plasma membrane of HepG2 cells, and this also was reduced by BAPTA. InsP3R2-mediated Ca2+ signals enhance organic anion secretion into bile by targeting Mrp2 to the canalicular membrane.
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DOI：
10.1016/j.bbrc.2004.07.192
发表时间：
2004-09-24
影响因子：
3.1
作者：
Correa, PRAV;Guerra, MT;Nathanson, MH
通讯作者：
Nathanson, MH
DOI：
10.1073/pnas.87.9.3557
发表时间：
1990-05-01
影响因子：
11.1
作者：
KITAMURA, T;JANSEN, P;ARIAS, IM
通讯作者：
ARIAS, IM
影响因子：
4.8
作者：
Minagawa, N;Kruglov, EA;Nathanson, MH
通讯作者：
Nathanson, MH
影响因子：
4.8
作者：
Berthiaume, F;Moghe, PV;Yarmush, ML
通讯作者：
Yarmush, ML
影响因子：
4.8
作者：
Hernandez, Erick;Leite, M. Fatima;Nathanson, Michael H.
通讯作者：
Nathanson, Michael H.