Upregulated acetylcholine synthesis during early differentiation in the embryonic stem cell line CGR8
Upregulated acetylcholine synthesis during early differentiation in the embryonic stem cell line CGR8
复制标题
胚胎干细胞系 CGR8 早期分化过程中乙酰胆碱合成上调
DOI：
10.1016/j.neulet.2013.04.052
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发表时间：
2013
影响因子：
2.5
通讯作者：
C. Kirkpatrick
中科院分区：
文献类型：
--
作者：
I. Wessler;R. Michel;H. Schmidt;S. Kaltwasser;R. Unger;C. Kirkpatrick
干细胞用于产生分化的体细胞，包括神经元细胞。研究了小鼠胚胎干细胞系CGR 8在早期分化过程中乙酰胆碱（一种神经递质和广泛表达的信号分子）的合成和释放，即在白血病抑制因子（LIF）存在下维持多能性和在LIF不存在下诱导早期分化。CGR 8细胞表达胆碱乙酰转移酶（ChAT），如通过测量酶活性和溴乙酰胆碱的实质性抑制所证明的。多能CGR 8细胞的ChAT活性为250 pmol乙酰胆碱/mg/h，每106个细胞含1.1pmol乙酰胆碱，每1 × 106个细胞6 h释放约12.00pmol乙酰胆碱。去除LIF诱导早期分化，如通过减少转录因子Oct-4和Nanog以及显著减慢增殖速率所证明的。在此条件下，乙酰胆碱合成增加至1640 pmol/mg/h;与多能状态相关，乙酰胆碱含量增加10倍，释放量约为32 pmol乙酰胆碱/1x 106细胞/6 h。酶动力学分析表明，前体乙酰辅酶A的Km和Vmax的显着增加，而前体胆碱的Km没有变化。总之，干细胞系CGR 8的早期分化与ChAT活性和乙酰胆碱释放的大幅增加有关。
Stem cells are used to generate differentiated somatic cells including neuronal cells. Synthesis and release of acetylcholine, a neurotransmitter and widely expressed signaling molecule, were investigated in the murine embryonic stem cell line CGR8 during early differentiation, i.e. in the presence of leukemia inhibitory factor (LIF) to maintain pluripotency and in the absence of LIF to induce early differentiation. CGR8 cells express choline acetyltransferase (ChAT) as demonstrated by measurement of enzyme activity and substantial inhibition by bromoacetylcholine. Pluripotent CGR8 cells showed a ChAT activity of 250pmol acetylcholine/mg/h, contained 1.1pmol acetylcholine/106cells and released about 12.00pmol acetylcholine/1x106cells/6h. Removal of LIF induced early differentiation as evidenced by reduced transcription factors Oct-4 and Nanog and a substantial slowing of the proliferation rate. Under this condition acetylcholine synthesis increased to 1640pmol/mg/h; related to the pluripotent state the content of acetylcholine increased 10-fold and the release to about 32pmol acetylcholine/1x106cells/6h. Enzyme kinetic analysis showed a significant increase of the Kmfor the precursor acetyl-CoA and of Vmaxwithout a change of the Kmfor the precursor choline. In conclusion, early differentiation of the stem cell line CGR8 is associated with a substantial increase in ChAT activity and acetylcholine release.
影响因子：
11.2
作者：
Song, Pingfang;Sekhon, Harmanjatinder S.;Spindel, Eliot R.
通讯作者：
Spindel, Eliot R.
毒蕈碱受体激动剂刺激基质金属蛋白酶1依赖性侵袭人结肠癌细胞。
DOI：
10.1016/j.bbrc.2011.10.052
发表时间：
2011-11-18
影响因子：
3.1
作者：
Raufman, Jean-Pierre;Cheng, Kunrong;Saxena, Neeraj;Chandi, Ahmed;Belo, Angelica;Khurana, Sandeep;Xie, Guofeng
通讯作者：
Xie, Guofeng