Effect of LIF-withdrawal on acetylcholine synthesis in the embryonic stem cell line CGR8 is not mediated by STAT3, PI3Ks or cAMP/PKA pathways.
Effect of LIF-withdrawal on acetylcholine synthesis in the embryonic stem cell line CGR8 is not mediated by STAT3, PI3Ks or cAMP/PKA pathways.
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LIF 撤除对胚胎干细胞系 CGR8 中乙酰胆碱合成的影响不是由 STAT3、PI3K 或 cAMP/PKA 途径介导的
DOI：
10.1016/j.intimp.2015.04.005
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发表时间：
2015
影响因子：
5.6
通讯作者：
Wessler I
中科院分区：
文献类型：
--
作者：
Michel-Schmidt R;Kirkpatrick CJ;Wessler I
乙酰胆碱（ACh）作为局部细胞信号分子，在自然界中广泛表达，包括哺乳动物细胞和胚胎干细胞。鼠胚胎干细胞系 CGR8 合成并释放大量的乙酰胆碱。特别是在早期分化期间（与基因/表型功能的多重改变相关的时期），ACh 的合成和释放增加了 10 倍。在小鼠干细胞中，STAT-3、PI3K 和 cAMP/PKA 途径的第二信使参与维持自我更新和多能性。本实验旨在测试这些信号通路的阻断剂是否在 LIF 存在的情况下（即 CGR8 具有多能性时）增强 ACh 细胞含量。 NSC74859 是 STAT-3 的抑制剂，既不影响增殖率，也不影响 ACh 细胞含量，而更敏感的 STAT-3 抑制剂 FLLL31 降低增殖率，并使 ACh 细胞含量增加约 3 倍。 PI3K抑制剂LY294002降低增殖率但不改变ACh细胞含量，而PKA抑制剂H89产生与FLLL31相当的效果。有趣的是，在对照实验中发现细胞密度和 ACh 细胞含量之间存在很强的负相关性，这可以解释在 FLLL31 和 H89 存在下观察到的 ACh 细胞含量增加 3 倍。 Forskolin（一种 PKA 激活剂）没有效果。总之，LIF 去除（即早期分化期间）诱导的 ACh 细胞含量增加 10 倍似乎不太可能是由 STAT-3、PI3K 和 cAMP/PKA 途径的第二信使介导的。然而，PI3K 途径似乎参与控制细胞密度和 ACh 细胞含量之间的反比关系，因为这种相关性在 LY294002 存在的情况下显着减弱。
Acetylcholine (ACh) acts as a local cellular signaling molecule and is widely expressed in nature, including mammalian cells and embryonic stem cells. The murine embryonic stem cell line CGR8 synthesizes and releases substantial amounts of ACh. Particularly during early differentiation – a period associated with multiple alterations in geno-/phenotype functions – synthesis and release of ACh are increased by 10-fold. In murine stem cells second messengers of the STAT-3, PI3K and cAMP/PKA pathways are involved in maintaining self-renewal and pluripotency. The present experiments were designed to test whether blockers of these signaling pathways enhance ACh cell content in the presence of LIF, i.e. when CGR8 is pluripotent. NSC74859, an inhibitor of STAT-3, affected neither the proliferation rate nor ACh cell content, whereas the more sensitive STAT-3 inhibitor FLLL31 reduced the proliferation rate and increased ACh cell content by about 3-fold. The PI3K inhibitor LY294002 reduced the proliferation rate but did not modify the ACh cell content, whereas the PKA inhibitor H89 produced effects comparable to FLLL31. Interestingly, in control experiments a strong inverse correlation was found between cell density and ACh cell content, which could explain the 3-fold increase in the ACh cell content observed in the presence of FLLL31 and H89. Forskolin, a PKA activator, had no effect. In conclusion, it appears unlikely that the 10-fold increase in ACh cell content induced by LIF removal, i.e. during early differentiation, is mediated by second messengers of the STAT-3, PI3K and cAMP/PKA pathways. However, the PI3K pathway appears to be involved in control of the inverse relation between cell density and ACh cell content, because this correlation was significantly attenuated in the presence of LY294002.
胚胎干细胞系 CGR8 早期分化过程中乙酰胆碱合成上调
影响因子：
2.5
作者：
I. Wessler;R. Michel;H. Schmidt;S. Kaltwasser;R. Unger;C. Kirkpatrick
通讯作者：
C. Kirkpatrick
影响因子：
6.1
作者：
Song, P;Sekhon, HS;Spindel, ER
通讯作者：
Spindel, ER
小鼠胚胎干细胞释放乙酰胆碱：烟碱和毒蕈碱受体的作用以及有机阳离子转运蛋白的阻断。
DOI：
--
发表时间：
2012
期刊：
Life Science
影响因子：
--
作者：
I. Wessler;R. Michel;E. Dohle;C. Kirkpatrick
通讯作者：
C. Kirkpatrick