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题名:L3/Lhx8 is a pivotal factor for cholinergic differentiation of murine embryonic stem cells
作者:T Manabe, K Tatsumi, M Inoue, M Makinodan, T Yamauchi, E Makinodan, S Yokoyama, R Sakumura, A Wanaka
来源:Cell Death and Differentiation[IF=7.785], (23 Feb 2007) Original Article
URL :http://dx.doi.org/10.1038/sj.cdd.4402106
日期:070330
摘要: T Manabe1,2, K Tatsumi1, M Inoue1, M Makinodan1, T Yamauchi1, E Makinodan1, S Yokoyama1, R Sakumura1 and A Wanaka1

1Department of 2nd Anatomy, Faculty of Medicine, Nara Medical University, 840 Shijyo-cho, Kasihara City, Nara 634-8521, Japan

Correspondence: T Manabe, Department of Neuronal Differentiation and Regeneration, Center for Developmental Biology (CDB), RIKEN Kobe Institute, 2-2-3 Minatojima-Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan. Tel: 81 78 306 3100; Fax: 81 78 306 3089; E-mail: manabe@cdb.riken.jp

2Current address: Department of Neural Differentiation and Regeneration, Center for Developmental Biology (CDB), RIKEN Kobe Institute, 2-2-3 Minatojima-Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan

Received 12 May 2006; Revised 22 December 2006; Accepted 4 January 2007; Published online 23 February 2007.

Abstract

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. Keywords: Lim-homeobox, L3/Lhx8, acetylcholine, GABA, RNAi Abbreviations: ES, embryonic stem; siRNA, small interfering RNA; MGE, medial ganglionic eminence; SDIA, stromal cell-derived inducing activity 101.

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