Induction of neurite outgrowth in PC12 cells treated with temperature-controlled repeated thermal stimulation.
Induction of neurite outgrowth in PC12 cells treated with temperature-controlled repeated thermal stimulation.
复制标题
DOI：
10.1371/journal.pone.0124024
复制
发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Izumi S
中科院分区：
文献类型：
--
作者：
Kudo TA;Kanetaka H;Mochizuki K;Tominami K;Nunome S;Abe G;Kosukegawa H;Abe T;Mori H;Mori K;Takagi T;Izumi S
为了促进损伤后神经系统的功能恢复，有必要提供能够诱导神经元再生活性，特别是神经突形成的最佳细胞外信号。本研究旨在研究温度控制的重复热刺激（TRTS）在大鼠PC 12嗜铬细胞瘤细胞中的神经突发生的调节，该细胞可被神经营养因子诱导分化为具有延长的神经突的神经元样细胞。使用加热板施加热刺激，并监测培养基温度与加热板的变化的表面温度的相关性。在生长或分化培养基中，通过加热板（加热板的预设表面温度，39.5°C或42°C）在两种不同温度下将平板化的PC 12细胞暴露于TRTS，每天最多18小时。然后，我们测量了生长，轴突发生或乙酰胆碱酯酶（AChE）活性（神经元标记物）的程度。为了分析TRTS对这些细胞的作用机制，我们使用以下物质检测了细胞内信号的变化：原肌球蛋白相关激酶A抑制剂GW 441756; p38丝裂原活化蛋白激酶（MAPK）抑制剂SB 203580; MAPK/细胞外信号调节激酶（ERK）激酶（MEK）抑制剂U 0126及其无活性类似物U 0124作为对照。虽然39.5°C的TRTS在细胞生长测定中没有降低细胞的生长速率，但在不添加其他神经突发生诱导剂的情况下，它确实增加了携带神经突的PC 12细胞的数量和AChE活性。此外，U 0126和SB 203580，而不是U 0124和GW 441756，显着抑制TRTS诱导的轴突发生。这些结果表明，TRTS可以诱导PC 12细胞中的神经突发生，并且ERK 12和p38 MAPK信号通路的参与是TRTS依赖的神经突发生所必需的。因此，TRTS可能是一种有效的再生神经医学技术。
To promote the functional restoration of the nervous system following injury, it is necessary to provide optimal extracellular signals that can induce neuronal regenerative activities, particularly neurite formation. This study aimed to examine the regulation of neuritogenesis by temperature-controlled repeated thermal stimulation (TRTS) in rat PC12 pheochromocytoma cells, which can be induced by neurotrophic factors to differentiate into neuron-like cells with elongated neurites. A heating plate was used to apply thermal stimulation, and the correlation of culture medium temperature with varying surface temperature of the heating plate was monitored. Plated PC12 cells were exposed to TRTS at two different temperatures via heating plate (preset surface temperature of the heating plate, 39.5°C or 42°C) in growth or differentiating medium for up to 18 h per day. We then measured the extent of growth, neuritogenesis, or acetylcholine esterase (AChE) activity (a neuronal marker). To analyze the mechanisms underlying the effects of TRTS on these cells, we examined changes in intracellular signaling using the following: tropomyosin-related kinase A inhibitor GW441756; p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580; and MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK) inhibitor U0126 with its inactive analog, U0124, as a control. While a TRTS of 39.5°C did not decrease the growth rate of cells in the cell growth assay, it did increase the number of neurite-bearing PC12 cells and AChE activity without the addition of other neuritogenesis inducers. Furthermore, U0126, and SB203580, but not U0124 and GW441756, considerably inhibited TRTS-induced neuritogenesis. These results suggest that TRTS can induce neuritogenesis and that participation of both the ERK1/2 and p38 MAPK signaling pathways is required for TRTS-dependent neuritogenesis in PC12 cells. Thus, TRTS may be an effective technique for regenerative neuromedicine.
登录
查看更多内容
影响因子：
2.1
作者：
Kudo, Tada-aki;Kanetaka, Hiroyasu;Hayashi, Haruhide
通讯作者：
Hayashi, Haruhide
影响因子：
5.4
作者：
El Omri, Abdelfatteh;Han, Junkyu;Isoda, Hiroko
通讯作者：
Isoda, Hiroko
影响因子：
2.9
作者：
Ahn, Tae-Beom;Jeon, Beom S.
通讯作者：
Jeon, Beom S.
影响因子：
2.2
作者：
Kudo, Tada-aki;Kanetaka, Hiroyasu;Hayashi, Haruhide
通讯作者：
Hayashi, Haruhide
影响因子：
4.8
作者：
Iwasaki, S;Hattori, A;Kohno, M
通讯作者：
Kohno, M