Fu Y M, Spirito P, Yu Z X, Biro S, Sasse J, Lei J, Ferrans V J, Epstein S E, Casscells W
Cardiology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
J Cell Biol. 1991 Sep;114(6):1261-73. doi: 10.1083/jcb.114.6.1261.
Compared to basic fibroblast growth factor (bFGF), a widely distributed, broad spectrum mitogen and mesoderm inducer, acidic fibroblast growth factor (aFGF) is reported to have an essentially neural distribution and to be undetectable in the early embryo. In the present investigation, we used immunoblotting and immunochemistry to assess the cellular and tissue distributions of aFGF and bFGF in 11-20-d rat embryos. Immunoblotting of crude and heparin-bound embryo extracts revealed faint bands at the expected 17-18-kD and predominant bands at an apparent molecular mass of 26 to 28-kD (despite reducing conditions) using multiple specific antibodies for aFGF and bFGF. Pretreatment with 8 M urea yielded 18-20-kD aFGF and bFGF and some 24-26-kD bFGF. Immunoreactivity for both aFGF and bFGF was positive and similar in the cytoplasm, nuclei, and extracellular matrix of cells of neuroectodermal and mesodermal origin, while it was negative in endoderm-derived cells. The distribution of immunoreactive aFGF and bFGF also showed changes during development that were associated with the process of cellular and tissue differentiation. For example, intensity and extent of immunoreactivity for both peptides progressively increased in the middle layer of the spinal cord with increasing differentiation of the neural cells. The immunostaining patterns were very similar for aFGF and bFGF for each organ and at each stage. In conclusion, high molecular mass forms of immunoreactive aFGF and bFGF are present in the rat embryo. Acidic FGF and bFGF are both widely distributed in tissues of neuroectodermal and mesodermal origin, and their distribution was very similar.
与广泛分布的广谱有丝分裂原和中胚层诱导剂碱性成纤维细胞生长因子(bFGF)相比,据报道酸性成纤维细胞生长因子(aFGF)主要分布于神经组织,在早期胚胎中无法检测到。在本研究中,我们使用免疫印迹法和免疫化学方法评估了11至20天龄大鼠胚胎中aFGF和bFGF的细胞和组织分布。使用多种针对aFGF和bFGF的特异性抗体,对粗制和肝素结合的胚胎提取物进行免疫印迹分析,结果显示,尽管处于还原条件下,但预期的17 - 18kD处出现淡带,表观分子量为26至28kD处出现主带。用8M尿素预处理可得到18 - 20kD的aFGF和bFGF以及一些24 - 26kD的bFGF。aFGF和bFGF在神经外胚层和中胚层来源细胞的细胞质、细胞核和细胞外基质中的免疫反应均为阳性且相似,而在内胚层来源的细胞中为阴性。免疫反应性aFGF和bFGF的分布在发育过程中也表现出变化,这些变化与细胞和组织分化过程相关。例如,随着神经细胞分化程度的增加,脊髓中层两种肽的免疫反应强度和范围逐渐增强。每个器官和每个阶段的aFGF和bFGF免疫染色模式非常相似。总之,大鼠胚胎中存在高分子量形式的免疫反应性aFGF和bFGF。酸性FGF和bFGF均广泛分布于神经外胚层和中胚层来源的组织中,且它们的分布非常相似。