Characterization of apical and basolateral plasma membrane domains derived from cultured rat cholangiocytes.
Characterization of apical and basolateral plasma membrane domains derived from cultured rat cholangiocytes.
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来自培养的大鼠胆管细胞的顶端和基底外侧质膜结构域的表征。
DOI：
10.1006/abio.1997.2431
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
LaRusso,NF
中科院分区：
文献类型：
--
作者：
Tietz,P;Levine,S;Holman,R;Fretham,C;LaRusso,NF
胆管细胞是排列在肝内胆管上的上皮细胞，由具有离散的顶端（内腔）和基底外侧结构域的质膜组成，这些结构域含有不同的通道、转运蛋白和受体。在最近的工作中，我们建立了正常大鼠胆管细胞（NRC）的长期原代培养系统。我们的目的是制备和表征顶端和基底侧质膜囊泡从NRC。使用连续等密度离心蔗糖梯度，我们产生了单独的顶端和基底侧质膜囊泡。我们通过透射电子显微镜、特异性标记酶测定和免疫印迹表征了这些囊泡;我们还使用生化方法确定了密封囊泡的百分比及其囊内体积和侧性。我们的研究结果表明，来自NRC的顶端和基底侧质膜结构域的囊泡是高度纯化的，主要是密封的，右侧出囊泡的大小和脂质和蛋白质组成不同。这个实验模型代表了一种新的工具，将是有用的额外的功能研究的通道，转运蛋白，和受体差异分布在胆管上皮细胞的质膜。
Cholangiocytes, the epithelial cells that line intrahepatic bile ducts, are composed of plasma membranes with discrete apical (lumenal) and basolateral domains that contain different channels, transporters, and receptors. In recent work, we developed a long-term, primary culture system of normal rat cholangiocytes (NRC). Our aims here were to prepare and characterize apical and basolateral plasma membrane vesicles from NRC. Using serial isopycnic centrifugation on sucrose gradients, we generated separate apical and basolateral plasma membrane vesicles. We characterized these vesicles by transmission electron microscopy, specific marker enzyme assays, and immunoblotting; we also determined the percentage of sealed vesicles and their intravesicular volume and sidedness using biochemical approaches. Our results showed that vesicles derived from the apical and basolateral plasma membrane domains of NRC were highly purified, predominately sealed, right-side-out vesicles that differed in size and lipid and protein composition. This experimental model represents a novel tool that will be useful for additional functional studies on the channels, transporters, and receptors differentially distributed in the plasma membrane of biliary epithelia.