Nakagami Y, Saito H, Matsuki N
Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan.
Jpn J Pharmacol. 1997 Dec;75(4):319-26. doi: 10.1254/jjp.75.319.
Neurotrophic effects in vitro have been generally related to promotion of differentiation, maturation and survival, but little is known about the effect on neuronal circuit formation. The organotypic culture system would be an available technique to investigate neuronal circuit formation and neuronal cell-cell interactions. As we reported previously, an optical recording system is a useful technique to comprehend neuronal activities and circuit from multi-points simultaneously. In this study, we investigated whether continuous application of basic fibroblast growth factor (bFGF or FGF-2) and brain-derived neurotrophic factor (BDNF) inhibited neuronal cell death induced by serum-deprivation in organotypic culture using propidium iodide staining, and we analyzed effects of bFGF and BDNF on the formation of neuronal circuits using the optical recording system. Continuous application of bFGF or BDNF significantly protected the slices from neuronal death. Optical recording also demonstrated that addition of 10 ng/ml bFGF or 50 ng/ml BDNF enhanced optical signals in all hippocampal areas significantly. These data strongly suggest that bFGF and BDNF promote the formation of neuronal circuits as well as survival and that optical recording of organotypic hippocampal slices would be a useful technique that enables us to analyze neuronal circuit formation easily.
体外神经营养作用一般与促进分化、成熟和存活有关，但对其在神经元回路形成方面的影响却知之甚少。器官型培养系统将是一种用于研究神经元回路形成和神经元细胞间相互作用的有效技术。正如我们之前所报道的，光学记录系统是一种从多个点同时理解神经元活动和回路的有用技术。在本研究中，我们使用碘化丙啶染色研究了在器官型培养中持续应用碱性成纤维细胞生长因子（bFGF 或 FGF - 2）和脑源性神经营养因子（BDNF）是否能抑制血清剥夺诱导的神经元细胞死亡，并且我们使用光学记录系统分析了 bFGF 和 BDNF 对神经元回路形成的影响。持续应用 bFGF 或 BDNF 能显著保护切片免受神经元死亡。光学记录还表明，添加 10 ng/ml 的 bFGF 或 50 ng/ml 的 BDNF 能显著增强所有海马区域的光学信号。这些数据有力地表明，bFGF 和 BDNF 不仅能促进神经元存活，还能促进神经元回路的形成，并且器官型海马切片的光学记录将是一种使我们能够轻松分析神经元回路形成的有用技术。
