Ishikawa R, Nishikori K, Furukawa S
National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Japan.
J Neurochem. 1991 Mar;56(3):836-41. doi: 10.1111/j.1471-4159.1991.tb01999.x.
We developed a sensitive two-site enzyme immunoassay (EIA) system for acidic fibroblast growth factor (aFGF), using a polyclonal antibody raised in rats. This assay is based on the sandwiching of the antigen between anti-aFGF antibody immunoglobulin G (IgG) coated on plates and biotinylated anti-aFGF antibody IgG; the detection of biotinylated IgG was performed by enzyme reaction of streptavidin-conjugated beta-D-galactosidase (beta-D-galactoside hydrolase; EC 3.2.1.23). Our system was specific for aFGF, because basic fibroblast growth factor, which shares a 55% homology of amino acid sequence with aFGF, hardly cross-reacted at all. The sensitivity of this system (0.2 ng/ml) enabled us to quantify endogenous immunoreactive aFGF in the CNS. Using this two-site EIA system, we examined the levels of aFGF in various regions of rat brain and their developmental changes. At the early stage of neonatal development, i.e., 2 days after birth, all brain regions registered low aFGF levels (less than 10 ng/g tissue). However, at the young adult stage (21- to 49-day-old animals), an extremely high level of aFGF (75-90 ng/g tissue) was found in the ponsmedulla; relatively high levels (30-40 ng/g tissue) were found in the diencephalon and mesencephalon; and comparatively low aFGF levels (5-15 ng/g tissue) were found in various other brain regions such as the frontal cortex, piriform cortex, hippocampus, olfactory bulb, cerebellum, and striatum. This marked change in the regional distribution of aFGF in the rat brain during postnatal development from 2 to 21 days after birth suggests that this factor plays a significant role in the brain during this period.
我们使用在大鼠中产生的多克隆抗体制备了一种用于酸性成纤维细胞生长因子（aFGF）的灵敏双位点酶免疫测定（EIA）系统。该测定基于抗原夹在包被在板上的抗aFGF抗体免疫球蛋白G（IgG）和生物素化抗aFGF抗体IgG之间；生物素化IgG的检测通过链霉亲和素结合的β-D-半乳糖苷酶（β-D-半乳糖苷水解酶；EC 3.2.1.23）的酶反应进行。我们的系统对aFGF具有特异性，因为与aFGF氨基酸序列有55%同源性的碱性成纤维细胞生长因子几乎没有交叉反应。该系统的灵敏度（0.2 ng/ml）使我们能够定量中枢神经系统中内源性免疫反应性aFGF。使用这种双位点EIA系统，我们检测了大鼠脑不同区域中aFGF的水平及其发育变化。在新生儿发育的早期阶段，即出生后2天，所有脑区的aFGF水平都很低（低于10 ng/g组织）。然而，在年轻成年阶段（21至49日龄动物），在脑桥延髓中发现了极高水平的aFGF（75至90 ng/g组织）；在间脑和中脑发现了相对较高的水平（30至40 ng/g组织）；在其他各种脑区，如额叶皮质、梨状皮质、海马体、嗅球、小脑和纹状体中发现了相对较低的aFGF水平（5至15 ng/g组织）。出生后2至21天大鼠脑内aFGF区域分布的这种显著变化表明该因子在此期间在脑中发挥着重要作用。
