Dini L, Kolb-Bachofen V
Eur J Cell Biol. 1987 Aug;44(1):144-50.
We investigated the number and distribution of galactose-specific binding sites in developing livers from suckling rats of various ages using Lac-BSA-Au5 (lactosylated bovine serum albumin adsorbed onto colloidal gold particles 5 nm in diameter) as electron-dense ligand, and performing transmission electron microscopy of the specimen. It has been reported that the number of galactose-specific binding sites increases rapidly during organ development post partum (p.p.) and this was ascribed to hepatocyte receptor increase only. We now have investigated in in situ and in vitro experiments whether the binding sites of identical sugar specificity but located on sinusoidal cells show the same increase in expression or are independently regulated. We therefore quantified the number of particles bound by isolated hepatocytes and liver macrophages and found a gradual increase of both binding activities with age, the binding levels of adult liver cells being reached at day 15 p.p. This was confirmed with experiments using in situ prefixed organs thus proving the validity of this finding also for the intact organ. In both sets of experiments--in vitro as well as in vivo--ligand was found binding statistically distributed as single particles on hepatocytes of all ages, whereas on liver macrophages the binding pattern changed during development. On liver macrophages from rats 15 days of age ligand binding occurs in the preclustered pattern described for macrophages from adult rat livers whereas liver macrophages of newborn rats express a different binding pattern: they bind the ligands mostly as single particles with only few and small microaggregates.(ABSTRACT TRUNCATED AT 250 WORDS)
我们使用Lac-BSA-Au5(吸附在直径5纳米的胶体金颗粒上的乳糖基化牛血清白蛋白)作为电子致密配体,对不同年龄的哺乳大鼠发育中的肝脏中半乳糖特异性结合位点的数量和分布进行了研究,并对标本进行了透射电子显微镜检查。据报道,产后(p.p.)器官发育过程中半乳糖特异性结合位点的数量迅速增加,这仅归因于肝细胞受体的增加。我们现在通过原位和体外实验研究了位于窦状细胞上的具有相同糖特异性的结合位点是否显示出相同的表达增加或受到独立调节。因此,我们对分离的肝细胞和肝巨噬细胞结合的颗粒数量进行了定量,发现两种结合活性均随年龄逐渐增加,成年肝细胞的结合水平在产后第15天达到。使用原位预固定器官的实验证实了这一点,从而也证明了这一发现对完整器官的有效性。在体外和体内这两组实验中,均发现配体以单颗粒形式统计学分布地结合在所有年龄的肝细胞上,而在肝巨噬细胞上,结合模式在发育过程中发生变化。在15日龄大鼠的肝巨噬细胞上,配体结合呈成年大鼠肝脏巨噬细胞所描述的预聚集模式,而新生大鼠的肝巨噬细胞表现出不同的结合模式:它们大多以单颗粒形式结合配体,只有少量小的微聚集体。(摘要截于250字)