Hanneken A, Lutty G A, McLeod D S, Robey F, Harvey A K, Hjelmeland L M
Wilmer Ophthalmological Institute, Johns Hopkins Hospital, Baltimore, Maryland 21205.
J Cell Physiol. 1989 Jan;138(1):115-20. doi: 10.1002/jcp.1041380116.
The basic fibroblast growth factor (FGF) is a potent mitogen that has vascular endothelium as one of its principle target cells. Recent work has provided both the complete amino acid sequence of basic FGF and the nucleotide sequence of the genes for both human and bovine basic FGF. Although capillary endothelial cells have been shown to produce basic FGF in vitro and to deposit basic FGF in their extracellular matrix in vitro as well, no direct evidence yet exists for the distribution of basic FGF in vivo. Antipeptide antibodies were prepared against a 15-amino-acid sequence from the amino terminus of basic FGF in order to avoid cross-reactivity with acidic FGF, a protein with 55% overall homology to basic FGF. After affinity purification, these antisera were used to localize the basic fibroblast growth factor in the fetal and adult bovine retina. Immunoreactive material was found in capillaries of the inner nuclear layer, a capillary network undergoing development during the third trimester in the fetal bovine eye. Although the resolution of the technique does not permit a unique assignment of cellular localization, the presence of stain immediately adjacent to the lumen of capillaries suggests that capillary endothelial cells may produce the basic fibroblast growth factor in vivo during vascular development.
碱性成纤维细胞生长因子(FGF)是一种有效的促细胞分裂剂,其主要靶细胞之一是血管内皮细胞。最近的研究工作已得出了碱性FGF的完整氨基酸序列以及人和牛碱性FGF基因的核苷酸序列。尽管已证明毛细血管内皮细胞在体外能产生碱性FGF,并且在体外也能将碱性FGF沉积于其细胞外基质中,但尚无碱性FGF在体内分布的直接证据。为了避免与酸性FGF(一种与碱性FGF总体同源性为55%的蛋白质)发生交叉反应,制备了针对碱性FGF氨基末端15个氨基酸序列的抗肽抗体。经过亲和纯化后,这些抗血清被用于在胎牛和成年牛视网膜中定位碱性成纤维细胞生长因子。在内核层的毛细血管中发现了免疫反应性物质,这是一个在胎牛眼妊娠晚期正在发育的毛细血管网络。尽管该技术的分辨率不允许对细胞定位进行唯一确定,但紧邻毛细血管管腔出现的染色表明,在血管发育过程中,毛细血管内皮细胞可能在体内产生碱性成纤维细胞生长因子。