Humpel C, Chadi G, Lippoldt A, Ganten D, Fuxe K, Olson L
Department of Histology, and Neurobiology, Karolinska Institute, Stockholm, Sweden.
Exp Brain Res. 1994;98(2):229-37. doi: 10.1007/BF00228411.
In vivo microdialysis is an established tool for sampling extracellular fluid compartments. However, microdialysis faces the problem that the implantation of the probe damages the microenvironment from which measurements are derived. In this study, we examined the expression of basic fibroblast growth factor mRNA and protein at the cellular level after implantation of a microdialysis probe into the dorsal hippocampus and found that 8 h after inserting the probe bFGF mRNA was markedly increased in a relatively large area centered around the probe, involving both the dorsal hippocampus and the overlying cerebral cortex, as revealed by radioactive in situ hybridization. Using nonradioactive in situ hybridization with digoxigenin-labelled riboprobes, combined with immunohistochemistry for glial fibrillary acidic protein we demonstrated that bFGF mRNA was exclusively increased in astrocytes at the probe insertion site. Using immunohistochemistry we also found that bFGF-like immunoreactivity was increased after implantation of the probe close to the lesion site, as shown by an increased number of bFGF immunoreactive nuclear glial profiles. These results provide evidence that the implantation of a microdialysis probe into the brain induces activation of bFGF gene expression in astrocytes associated with nuclear bFGF-like immunoreactivity. We conclude that lesion-induced effects have to be considered when evaluating microdialysis data, and that mechanical trauma to the brain will activate astroglial trophism, as seen from the increased density of astroglial profiles demonstrating bFGF mRNA and protein levels.
体内微透析是一种用于采集细胞外液区室样本的成熟工具。然而,微透析面临着一个问题,即探针植入会破坏用于测量的微环境。在本研究中,我们将微透析探针植入背侧海马体后,在细胞水平上检测了碱性成纤维细胞生长因子(bFGF)mRNA和蛋白的表达,发现插入探针8小时后,通过放射性原位杂交显示,在以探针为中心的相对较大区域(包括背侧海马体和上方的大脑皮层),bFGF mRNA显著增加。使用地高辛标记的核糖探针进行非放射性原位杂交,并结合胶质纤维酸性蛋白的免疫组织化学,我们证明bFGF mRNA仅在探针插入部位的星形胶质细胞中增加。通过免疫组织化学我们还发现,在靠近损伤部位植入探针后,bFGF样免疫反应性增加,表现为bFGF免疫反应性核胶质细胞轮廓数量增加。这些结果提供了证据,表明将微透析探针植入大脑会诱导与核bFGF样免疫反应性相关的星形胶质细胞中bFGF基因表达的激活。我们得出结论,在评估微透析数据时必须考虑损伤诱导的效应,并且从显示bFGF mRNA和蛋白水平的星形胶质细胞轮廓密度增加可以看出,对大脑的机械损伤会激活星形胶质细胞的营养作用。