Rudland P S, Platt-Higgins A M, Wilkinson M C, Fernig D G
Department of Biochemistry, University of Liverpool, United Kingdom.
J Histochem Cytochem. 1993 Jun;41(6):887-98. doi: 10.1177/41.6.7686196.
We raised antiserum to human recombinant basic fibroblast growth factor (rbFGF) in rabbits. With this affinity-purified antiserum, other antisera to rbFGF, and commercial antiserum to bovine pituitary bFGF, we undertook immunocytochemical detection of bFGF in histological sections of rat mammary glands at different developmental stages. In non-growing ducts, anti-bFGF serum stains the basement membrane/myoepithelial cells, whereas in serial sections most of this stain is observed to be associated with anti-laminin-staining basement membranes rather than with anti-callus-keratin-staining myoepithelial cells. The weak staining of the myoepithelial cells is enhanced when NiCl2 is included in the detection system, but little staining for bFGF is observed in the epithelial cells. In growing neonatal ducts from 1-day-old rats, in growing terminal end buds (TEBs) and, to a lesser extent, in growing alveolar buds (ABs) in prepubescent (21-day) and pubescent (50-day) rats, both their inner and outer cells are stained moderately by anti-bFGF sera. In non-growing ducts from rats aged 6 days, in non-growing ABs of rats aged 60 days and more, and in alveoli from pregnant and lactating rats, only the basement membrane/myoepithelial cell area is stained by anti-bFGF sera; the epithelial cells are unstained. Staining of the myoepithelial cells is enhanced by mixtures of rbFGF and anti-bFGF sera in non-growing ducts, but there is little change in the staining of growing TEBs. All staining by anti-bFGF sera is abolished with heparin in the reactions. We suggest that the immunoreactive bFGF is present mainly bound to heparan sulfate glycosaminoglycans in the basement membrane of resting structures, but that immunoreactive bFGF becomes associated with proliferating cells, particularly those intermediate in characteristics between epithelial and myoepithelial cells in growing structures such as TEBs.
我们在兔体内制备了针对人重组碱性成纤维细胞生长因子（rbFGF）的抗血清。利用这种亲和纯化的抗血清、其他针对rbFGF的抗血清以及针对牛垂体bFGF的商业抗血清，我们对不同发育阶段大鼠乳腺组织切片中的bFGF进行了免疫细胞化学检测。在未生长的导管中，抗bFGF血清可使基底膜/肌上皮细胞染色，而在连续切片中，大部分这种染色被观察到与抗层粘连蛋白染色的基底膜相关，而非与抗角蛋白染色的肌上皮细胞相关。当检测系统中加入氯化镍时，肌上皮细胞的弱染色会增强，但上皮细胞中几乎未观察到bFGF染色。在1日龄大鼠的生长中的新生导管、生长中的终末芽（TEB）以及在较小程度上在青春期前（21日龄）和青春期（50日龄）大鼠的生长中的肺泡芽（AB）中，抗bFGF血清可使它们的内、外细胞适度染色。在6日龄大鼠的未生长导管、60日龄及以上大鼠的未生长AB以及妊娠和哺乳期大鼠的肺泡中，抗bFGF血清仅使基底膜/肌上皮细胞区域染色；上皮细胞未染色。在未生长的导管中，rbFGF和抗bFGF血清的混合物可增强肌上皮细胞的染色，但生长中的TEB的染色几乎没有变化。抗bFGF血清的所有染色在反应中都可被肝素消除。我们认为，免疫反应性bFGF主要以结合于静止结构基底膜中的硫酸乙酰肝素糖胺聚糖的形式存在，但免疫反应性bFGF会与增殖细胞相关，特别是在生长结构如TEB中具有上皮和肌上皮细胞之间中间特征的那些细胞。
