Bartles J R, Hubbard A L
Dev Biol. 1986 Nov;118(1):286-95. doi: 10.1016/0012-1606(86)90095-3.
We have utilized antibodies against five domain-specific integral proteins of the rat hepatocyte plasma membrane to examine the fates of the plasma membrane domains during hepatocyte division in the regenerating rat liver. The proteins were quantified on immunoblots of liver homogenates prepared during the peak of hepatocyte mitotic activity, 28-30 hr after two-thirds hepatectomy. Two sinusoidal/lateral proteins, CE 9 and the asialoglycoprotein receptor, and one bile canalicular protein, dipeptidylpeptidase IV, were not changed significantly in amount; whereas one sinusoidal/lateral protein, the epidermal growth factor receptor, and one bile canalicular protein, HA 4, were reduced to less than or equal to 50% of control levels. Light microscopic examination of plastic sections of regenerating liver tissue revealed that the mitotic hepatocytes generally appeared to retain normal contacts with neighboring interphase hepatocytes. Immunofluorescence was used to localize the domain-specific proteins on mitotic hepatocytes identified in 0.5-micron frozen sections of 28- to 30-hr regenerating liver tissue. Independent of mitotic stage, the hepatocytes retained mutually exclusive bile canalicular and sinusoidal/lateral domains, as defined at the molecular level by the distributions of specific proteins, such as HA 4 and CE 9, respectively.
我们利用针对大鼠肝细胞质膜五种结构域特异性整合蛋白的抗体,来研究再生大鼠肝脏中肝细胞分裂过程中质膜结构域的命运。在三分之二肝切除术后28 - 30小时肝细胞有丝分裂活动高峰期制备的肝脏匀浆免疫印迹上对这些蛋白质进行定量。两种窦状隙/侧面蛋白,即CE 9和去唾液酸糖蛋白受体,以及一种胆小管蛋白,二肽基肽酶IV,其含量没有显著变化;而一种窦状隙/侧面蛋白,即表皮生长因子受体,和一种胆小管蛋白,HA 4,减少至对照水平的50%或更低。对再生肝组织塑料切片的光镜检查显示,有丝分裂的肝细胞通常似乎与相邻的间期肝细胞保持正常接触。免疫荧光用于在28至30小时再生肝组织的0.5微米冰冻切片中鉴定的有丝分裂肝细胞上定位结构域特异性蛋白。与有丝分裂阶段无关,肝细胞保留了相互排斥的胆小管和窦状隙/侧面结构域,分别由特定蛋白如HA 4和CE 9的分布在分子水平上定义。