Matsuura S, Eto S, Kato K, Tashiro Y
J Cell Biol. 1984 Jul;99(1 Pt 1):166-73. doi: 10.1083/jcb.99.1.166.
Rat livers were prefixed by perfusion with 0.6% glutaraldehyde and briefly homogenized with a Teflon-glass homogenizer. The prefixed cells isolated by low-speed centrifugation in high yield effectively preserved the original polygonal shape and polarity. These cells were incubated with ferritin-antibody conjugates monospecific for rat liver 5'-nucleotidase, and the localization of the enzymes on the surface of hepatocytes and endothelial cells was quantitatively investigated. It was revealed that the surface density of 5'-nucleotidase is much higher on the bile canalicular surface than on the sinusoidal surface and only a few ferritin particles were detected on the lateral surface. On the bile canalicular surface ferritin particles were almost exclusively found on the microvilli in larger clusters. Similar distribution was also observed on the sinusoidal surface but the size of cluster was much smaller. On both surfaces many fewer ferritin particles were found on the intermicrovillar region, including the coated pits region, than on the microvillar region. Ferritin particles were also found on the endothelial cell surface.
大鼠肝脏先用0.6%的戊二醛灌注固定，然后用聚四氟乙烯-玻璃匀浆器进行短暂匀浆。通过低速离心高产率分离得到的固定细胞有效地保留了原始的多边形形状和极性。将这些细胞与对大鼠肝脏5'-核苷酸酶具有单特异性的铁蛋白-抗体偶联物孵育，并对酶在肝细胞和内皮细胞表面的定位进行定量研究。结果显示，5'-核苷酸酶在胆小管表面的表面密度远高于窦状隙表面，在侧面仅检测到少量铁蛋白颗粒。在胆小管表面，铁蛋白颗粒几乎只在较大簇的微绒毛上发现。在窦状隙表面也观察到类似的分布，但簇的大小要小得多。在两个表面上，包括被膜小窝区域在内的微绒毛间区域发现的铁蛋白颗粒比微绒毛区域少得多。在内皮细胞表面也发现了铁蛋白颗粒。
