LKB1/AMPK and PKA control ABCB11 trafficking and polarization in hepatocytes.
LKB1/AMPK and PKA control ABCB11 trafficking and polarization in hepatocytes.
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DOI：
10.1371/journal.pone.0091921
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Arias IM
中科院分区：
文献类型：
--
作者：
Homolya L;Fu D;Sengupta P;Jarnik M;Gillet JP;Vitale-Cross L;Gutkind JS;Lippincott-Schwartz J;Arias IM
肝细胞的极化表现为胆小管网络的形成和控制细胞能量代谢的LKB1和AMPK的激活。胆汁酸，牛磺酸胆酸盐，也通过激活AMPK调节小管网络的发育。在本研究中，我们使用对照小鼠和肝脏特异性LKB1敲除小鼠的胶原夹心肝细胞培养物来检测LKB1在小管胆汁酸转运体ABCB11运输中的作用。在极化肝细胞中，ABCB11从高尔基体转运到顶质膜，并通过rab 11a-肌球蛋白Vb循环内体系统进行内源性循环。LKB1基因敲除小鼠出现黄疸、体重减轻、胆小管形成和ABCB11细胞内转运受损，并在三周内死亡。通过活细胞成像、光漂白后荧光恢复（FRAP）、颗粒跟踪和生物化学，我们发现LKB1活性是ABCB11微管依赖性运输到管膜所必需的。在对照肝细胞中，牛磺胆酸盐和cAMP可加速ABCB11的转运；然而，在LKB1基因敲除的肝细胞中，ABCB11向根尖膜的转运仅被cAMP而不是牛磺胆酸盐大大减少和恢复。cAMP通过pka介导的途径起作用，而不激活AMPK。我们的研究确定了LKB1在ABCB11向小管膜转运、肝细胞极化和小管网络形成中的调节作用。
Polarization of hepatocytes is manifested by bile canalicular network formation and activation of LKB1 and AMPK, which control cellular energy metabolism. The bile acid, taurocholate, also regulates development of the canalicular network through activation of AMPK. In the present study, we used collagen sandwich hepatocyte cultures from control and liver-specific LKB1 knockout mice to examine the role of LKB1 in trafficking of ABCB11, the canalicular bile acid transporter. In polarized hepatocytes, ABCB11 traffics from Golgi to the apical plasma membrane and endogenously cycles through the rab 11a-myosin Vb recycling endosomal system. LKB1 knockout mice were jaundiced, lost weight and manifested impaired bile canalicular formation and intracellular trafficking of ABCB11, and died within three weeks. Using live cell imaging, fluorescence recovery after photobleaching (FRAP), particle tracking, and biochemistry, we found that LKB1 activity is required for microtubule-dependent trafficking of ABCB11 to the canalicular membrane. In control hepatocytes, ABCB11 trafficking was accelerated by taurocholate and cAMP; however, in LKB1 knockout hepatocytes, ABCB11 trafficking to the apical membrane was greatly reduced and restored only by cAMP, but not taurocholate. cAMP acted through a PKA-mediated pathway which did not activate AMPK. Our studies establish a regulatory role for LKB1 in ABCB11 trafficking to the canalicular membrane, hepatocyte polarization, and canalicular network formation.
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影响因子：
--
作者：
Hawley SA;Boudeau J;Reid JL;Mustard KJ;Udd L;Mäkelä TP;Alessi DR;Hardie DG
通讯作者：
Hardie DG
影响因子：
4.9
作者：
Swift B;Brouwer KL
通讯作者：
Brouwer KL
DOI：
10.1042/bj20101721
发表时间：
2011-02-15
期刊：
The Biochemical journal
影响因子：
--
作者：
Woods A;Heslegrave AJ;Muckett PJ;Levene AP;Clements M;Mobberley M;Ryder TA;Abu-Hayyeh S;Williamson C;Goldin RD;Ashworth A;Withers DJ;Carling D
通讯作者：
Carling D
影响因子：
56.9
作者：
Shaw, RJ;Lamia, KA;Cantley, LC
通讯作者：
Cantley, LC
DOI：
10.1073/pnas.1120421109
发表时间：
2012-02-14
影响因子：
11.1
作者：
Partanen, Johanna I.;Tervonen, Topi A.;Klefstrom, Juha
通讯作者：
Klefstrom, Juha