Treatment with deferoxamine increases neurons from neural stem/progenitor cells
Treatment with deferoxamine increases neurons from neural stem/progenitor cells
复制标题
DOI：
10.1016/j.brainres.2006.02.046
复制
发表时间：
2006-05-30
期刊：
影响因子：
2.9
通讯作者：
Nishino, Hitoo
中科院分区：
文献类型：
--
作者：
Kim, Hye-Jung;Hida, Hideki;Nishino, Hitoo
神经移植是治疗神经退行性疾病的一种很有前途的方法。神经干/祖细胞（NPC）具有自我更新和多能性，因此当它们被明确定义为分化为神经元时，它们是供体细胞的良好候选者。由于神经元分化遵循细胞周期退出，我们研究了用细胞周期阻断剂处理是否增加NPC的神经元产生。取E12.5大鼠中脑腹侧神经前体细胞，在含氮和碱性成纤维细胞生长因子的DMEM/F12培养基中培养成神经球。去铁胺（G1/S期阻滞剂）处理NPC后，分化后β-微管蛋白III阳性细胞的数量增加，伴随着MAP 2 mRNA和蛋白的增加以及GFAP蛋白的减少。此外，证实了β-微管蛋白III/BrdU双阳性细胞的增加和GFAP/BrdU双阳性细胞的减少。在实时荧光定量PCR中，p21（cip 1），p27（kip 1）和p57（kip 2）mRNA的表达在去铁胺处理后8小时内保持不变，但在1天后显著升高。去铁胺可特异性地促进p27（kip 1）mRNA在1-2 d的表达和p27（kip 1）蛋白在3 d的表达，沿着neuroD启动子的激活和neuroD mRNA的表达。p27（kip 1）转染入NPC诱导neuroD启动子的激活和β-微管蛋白III阳性细胞数量的增加。这些数据表明，去铁胺预处理增加了与p27（kip 1）升高和neuroD信号通路激活相关的NPC神经元数量。以这种方式，细胞周期的调节应该是一个有用的第一步，在神经移植工程的NPC。(c)2006 Elsevier B. V.保留所有权利。
Neural transplantation is a promising approach for treating neurodegenerative disease. Neural stem/progenitor cells (NPCs) are self-renewing and multipotent and thus are good candidates for donor cells when they have been clearly defined to differentiate into neurons. As neuronal differentiation follows cell cycle exit, we investigated whether neuron production from NPCs is increased by treatment with cell cycle blockers. NPCs from E12.5 rat ventral mesencephalon were cultured as neurospheres in DMEM/F12 medium containing N2 supplements and bFGF. Treatment of NPCs with deferoxamine, a G1/S phase blocker, increased the number of beta-tubulin III-positive cells after differentiation, concomitant with increases of MAP2 mRNA and protein, and a decrease of GFAP protein. Further, an increase in beta-tubulin III/BrdU double-positive cells and a decrease in GFAP/BrdU double-positive cells were confirmed. In real-time PCR, the expressions of p21(cip1), p27(kip1) and p57(kip2) mRNAs remained unaltered for 8 h after treatment with deferoxamine but were significantly elevated after I day. Deferoxamine specifically enhanced the elevation of p27(kip1) mRNA at 1-2 days and the accumulation of p27(kip1) protein at 3 days, along with the activation of neuroD promoter and the elevation of neuroD mRNA. Transfection of p27(kip1) into NPCs induced activation of neuroD promoter and increase of number of beta-tubulin III-positive cells. These data suggest that pretreatment with deferoxamine increases the number of neurons from NPCs related to prolonged p27(kip1) elevation and activation of the neuroD signaling pathway. In this way, regulation of the cell cycle should be a useful first step in engineering NPCs for neural transplantation. (c) 2006 Elsevier B.V. All rights reserved.