Department of Anatomy, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-gu, Seoul 120-752, South Korea.
Life Sci. 2011 Sep 26;89(13-14):439-49. doi: 10.1016/j.lfs.2011.07.003. Epub 2011 Jul 27.
Our study aimed to demonstrate whether agmatine (Ag) could regulate proliferation and cell fate determination of subventricular zone neural stem cells (SVZ NSCs).
SVZ NSCs were grown in the presence of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) (20ng/ml) until 4days in vitro (DIV) and later the culture medium was replaced without EGF and bFGF until 11 DIV in the absence (EGF/bFGF(+/-)/Ag(-)) or presence of agmatine (EGF/bFGF(+/-)/Ag(+)). Another set SVZ NSCs were maintained with EGF and bFGF until 11 DIV without (EGF/bFGF(+/+)/Ag(-)) or with agmatine treatment (EGF/bFGF(+/+)/Ag(+)). Agmatine's effect on proliferation and cell death (H and PI staining and Caspase-3 immunostaining) was examined at DIV 4 and 11. Agmatine's (100μM) effect on cell fate determination was confirmed by immunostaining and Western blot at 11 DIV.
Agmatine treatment reduced the neurosphere size and total cell count number dose-dependently in all the experimental groups both at DIV 4 and11. Immunoblotting and staining results showed that agmatine increased the Tuj1 and Microtubule-associated protein 2 (MAP2) and decreased the Glial fibrillary acidic protein (GFAP) with no change in the Oligo2 protein expressions. This neurogenesis effect of agmatine seems to have a relation with Extracellular-signal-regulated kinases (ERK1/2) activation and anti-astrogenesis effect is thought to be related with the suppression of Bone morphogenetic proteins (BMP) 2,4 and contraction of Sma and Mad (SMAD) 1,5,8 protein expression.
This model could be an invaluable tool to study whether agmatine treated SVZ NSC transplantation to the central nervous system (CNS) injury could trigger neurogenesis and decrypt the full range of molecular events involved during neurogenesis in vivo as evidenced in vitro.
本研究旨在探讨胍丁胺（Agmatine）是否能够调节侧脑室下区神经干细胞（SVZ NSCs）的增殖和细胞命运决定。
将 SVZ NSCs 在表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）（20ng/ml）存在的情况下培养至体外 4 天（DIV），然后在无 EGF 和 bFGF 的情况下（EGF/bFGF(+/-)/Ag(-)）或存在胍丁胺的情况下（EGF/bFGF(+/-)/Ag(+))继续培养至 11 DIV。另一组 SVZ NSCs 在 EGF 和 bFGF 的存在下培养至 11 DIV，无胍丁胺处理（EGF/bFGF(+/+)/Ag(-)）或有胍丁胺处理（EGF/bFGF(+/+)/Ag(+))。在 DIV 4 和 11 时，通过 H 和 PI 染色和 caspase-3 免疫染色检测胍丁胺对增殖和细胞死亡的影响。在 11 DIV 时，通过免疫染色和 Western blot 验证胍丁胺对细胞命运决定的影响。
在所有实验组中，胍丁胺处理均呈剂量依赖性地减少神经球的大小和总细胞计数，无论是在 DIV 4 还是 11 时。免疫印迹和染色结果表明，胍丁胺增加了 Tuj1 和微管相关蛋白 2（MAP2）的表达，减少了胶质纤维酸性蛋白（GFAP）的表达，而少突胶质细胞 2（Oligo2）蛋白的表达没有变化。胍丁胺的这种神经发生效应似乎与细胞外信号调节激酶（ERK1/2）的激活有关，而抗星形胶质细胞效应则与骨形态发生蛋白（BMP）2、4 的抑制和 Smad1、5、8 蛋白表达的收缩有关。
该模型可能是研究胍丁胺处理的 SVZ NSC 移植到中枢神经系统（CNS）损伤是否能触发神经发生，并解密体内神经发生过程中涉及的一系列分子事件的宝贵工具，这在体外得到了证实。
