Westermann R, Johannsen M, Unsicker K, Grothe C
Department of Anatomy and Cell Biology, University of Marburg, F.R.G.
J Neurochem. 1990 Jul;55(1):285-92. doi: 10.1111/j.1471-4159.1990.tb08850.x.
Basic fibroblast growth factor (bFGF) has recently been isolated from bovine adrenal glands. Immunohistological data revealed its presence in both adrenal cortex and adrenal medulla. Using immuno-electronmicroscopy, we found that in medullary chromaffin cells bFGF-immunoreactivity is localized in the secretory granules. Immunoreactivity also was observed by electronmicroscopy in isolated granules. Western blot analysis revealed the presence of the typical 18-kDa bFGF and additional immunoreactive materials with molecular masses of approximately 24, 30, and 46 kDa in whole bovine adrenal, and in cortex and medulla. Similar results were obtained with proteins from bovine chromaffin granules, with the following two exceptions: the 46-kDa immunoreactivity was found to be highly enriched when compared with medulla or cortex, and the 18-kDa band could be detected with only an antiserum against a synthetic peptide comprising the 24 NH2-terminal amino acids of bFGF, and not with an antiserum against purified bovine pituitary bFGF. All fractions enriched for bFGF-immunoreactivity showed neurotrophic activity for chick ciliary ganglion neurons, which could be blocked by antibodies. These results demonstrate for the first time the localization and occurrence of bFGF in a cellular secretory organelle, and present further evidence for the existence of higher molecular weight immunoreactive forms of bFGF.
碱性成纤维细胞生长因子(bFGF)最近已从牛肾上腺中分离出来。免疫组织学数据显示其存在于肾上腺皮质和肾上腺髓质中。利用免疫电子显微镜技术,我们发现,在髓质嗜铬细胞中,bFGF免疫反应性定位于分泌颗粒中。在分离出的颗粒中通过电子显微镜也观察到了免疫反应性。蛋白质印迹分析显示,在整个牛肾上腺、肾上腺皮质和髓质中均存在典型的18 kDa bFGF以及分子量约为24、30和46 kDa的其他免疫反应性物质。从牛嗜铬颗粒蛋白中也获得了类似结果,但有以下两个例外:与髓质或皮质相比,46 kDa免疫反应性物质高度富集;18 kDa条带仅能用针对包含bFGF 24个NH2末端氨基酸的合成肽的抗血清检测到,而不能用针对纯化的牛垂体bFGF的抗血清检测到。所有富含bFGF免疫反应性的组分对鸡睫状神经节神经元均表现出神经营养活性,且这种活性可被抗体阻断。这些结果首次证明了bFGF在细胞分泌细胞器中的定位和存在,并为存在更高分子量的bFGF免疫反应性形式提供了进一步证据。