L3/Lhx8 is a pivotal factor for cholinergic differentiation of murine embryonic stem cells
L3/Lhx8 is a pivotal factor for cholinergic differentiation of murine embryonic stem cells
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DOI：
10.1038/sj.cdd.4402106
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发表时间：
2007-06-01
影响因子：
12.4
通讯作者：
Wanaka, A.
中科院分区：
文献类型：
--
作者：
Manabe, T.;Tatsumi, K.;Wanaka, A.
L3/Lhx8是LIM-homeobox基因家族的成员。先前，我们证明L3/Lhx8-缺失小鼠在基底前脑特异性地缺乏胆碱能神经元。在本研究中，我们使用由H1.2启动子驱动的载体产生的L3/Lhx8靶向小干扰RNA (siRNA)，有条件地抑制了视黄酸诱导的小鼠胚胎干（ES）细胞系神经分化过程中L3/Lhx8的功能。我们的培养条件诱导胚胎干细胞有效分化为神经元和星形胶质细胞，但分化为少突胶质细胞的效率要低得多。通过siRNA抑制L3/Lhx8的表达导致胆碱能标记物ChAT阳性的细胞数量急剧减少，过表达L3/Lhx8恢复了这一作用。而Tuj1 +神经元和GABA +细胞数量未见明显变化。这些结果强烈提示L3/Lhx8是小鼠ES细胞胆碱能分化的关键因子，并参与基底前脑发育。
L3/Lhx8 is a member of the LIM-homeobox gene family. Previously, we demonstrated that L3/Lhx8- null mice specifically lacked cholinergic neurons in the basal forebrain. In the present study, we conditionally suppressed L3/Lhx8 function during retinoic acid-induced neural differentiation of a murine embryonic stem (ES) cell line using an L3/Lhx8-targeted small interfering RNA ( siRNA) produced by an H1.2 promoter-driven vector. Our culture conditions induced efficient differentiation of the ES cells into neurons and astrocytes, but far less efficient differentiation into oligodendrocytes. Suppression of L3/Lhx8 expression by siRNA led to a dramatic decrease in the number of cells positive for the cholinergic marker ChAT, and overexpression of L3/Lhx8 recovered this effect. However, no significant changes were observed in the number of Tuj1 + neurons and GABA + cells. These results strongly suggest that L3/Lhx8 is a key factor in the cholinergic differentiation of murine ES cells and is involved in basal forebrain development.