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Differential basolateral–apical distribution of scavenger receptor class B type I in cultured cells and the liver

机译:清道夫受体在培养细胞和肝脏中的B类I类清除剂的基底基底-顶端分布差异

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摘要

The high-density lipoprotein (HDL) receptor, scavenger receptor class B, type I (SR-BI), mediates selective cholesteryl ester uptake into the liver, which finally results in cholesterol secretion into the bile. Despite several reports, the distribution of hepatic SR-BI between the sinusoidal and canalicular membranes is still under debate. We present immunohistological data using specific markers showing that the bulk of SR-BI is present in sinusoidal membranes and, to a lesser extent, in canalicular membranes in murine and human liver sections. In addition, SR-BI was detected in preparations of rat liver canalicular membranes. We also compared the in vivo findings to HepG2 cells, a widely used in vitro hepatocyte model. Interestingly, SR-BI was enriched in bile canalicular-like (BC-like) structures in polarized HepG2 cells, which were cultivated either conventionally to form a monolayer or in Matrigel to form three-dimensional structures. Fluorescently labeled HDL was transported into close proximity of BC-like structures, whereas HDL labeled with the fluorescent cholesterol analog BODIPY-cholesterol was clearly detected within these structures. Importantly, similarly to human and mouse liver, SR-BI was localized in basolateral membranes in three-dimensional liver microtissues from primary human liver cells. Our results demonstrate that SR-BI is highly enriched in sinusoidal membranes and is also found in canalicular membranes. There was no significant basolateral–apical redistribution of hepatic SR-BI in fasting and refeeding experiments in mice. Furthermore, in vitro studies in polarized HepG2 cells showed explicit differences as SR-BI was highly enriched in BC-like structures. These structures are, however, functional and accumulated HDL-derived cholesterol. Thus, biological relevant model systems should be employed when investigating SR-BI distribution in vitro.Electronic supplementary materialThe online version of this article (doi:10.1007/s00418-014-1251-9) contains supplementary material, which is available to authorized users.
机译:高密度脂蛋白(HDL)受体,B类清道夫受体,I型(SR-BI),介导肝脏选择性摄取胆固醇酯,最终导致胆固醇分泌到胆汁中。尽管有几篇报道,但肝SR-BI在正弦膜和小管膜之间的分布仍在争论中。我们使用特定的标记物提供的免疫组织学数据表明,大部分SR-BI存在于正弦膜中,并且在较小程度上存在于鼠类和人类肝脏切片的小管膜中。此外,在大鼠肝小管膜制剂中检测到SR-BI。我们还将体内发现与广泛使用的体外肝细胞模型HepG2细胞进行了比较。有趣的是,SR-BI在极化的HepG2细胞中富集了胆小管状(BC状)结构,可按常规方式培养以形成单层或在基质胶中培养以形成三维结构。荧光标记的HDL被转运到BC样结构的附近,而在这些结构中清楚地检测到了被荧光胆固醇类似物BODIPY-胆固醇标记的HDL。重要的是,类似于人和小鼠的肝脏,SR-BI位于原代人肝细胞的三维肝脏显微组织的基底外侧膜中。我们的结果表明,SR-BI在正弦膜中高度富集,在小管膜中也有发现。在小鼠的禁食和再喂养实验中,肝SR-BI没有明显的基底外侧-根尖重新分布。此外,对极化的HepG2细胞的体外研究显示出明显的差异,因为SR-BI高度富含BC样结构。但是,这些结构是功能性和累积性HDL衍生的胆固醇。因此,在体外研究SR-BI分布时,应使用生物学相关的模型系统。电子补充材料本文的在线版本(doi:10.1007 / s00418-014-1251-9)包含补充材料,授权用户可以使用。

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