Knockdown of the L3/Lhx8 gene suppresses cholinergic differentiation of murine embryonic stem cell-derived spheres.
L3/Lhx8, a member of the Lim-homeobox gene family, is selectively and specifically expressed in the murine embryonic medial ganglionic eminence (MGE). Our previous study demonstrated that L3/Lhx8-deficient mice specifically lack cholinergic neurons in the basal forebrain. In this manuscript, we report the in vitro effects of reduced L3/Lhx8 gene expression on cholinergic differentiation in murine embryonic stem (ES) cell-derived spheres without dissociation. The knockdown of L3/Lhx8 gene expression dramatically decreased the cholinergic phenotype of spheres without altering other known phenotypes (TuJ1, GABA and GFAP). These results strongly suggest that L3/Lhx8 is a key factor for cholinergic differentiation of murine ES cell-derived spheres and is involved in basal forebrain development.
Manabe
Takayuki
T
Department of 2nd Anatomy, Faculty of Medicine, Nara Medical University, 840 Shijyo-cho, Kasihara City, Nara 634-8521, Japan. manabe@cdb.riken.jp
Tatsumi
Kouko
K
Inoue
Masahide
M
Matsuyoshi
Hiroko
H
Makinodan
Manabu
M
Yamauchi
Takahira
T
Makinodan
Eri
E
Yokoyama
Shohei
S
Sakumura
Ryoko
R
Okuda
Hiroaki
H
Wanaka
Akio
A
D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D020532:Basal Nucleus of Meynert / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D002799:Cholinergic Fibers / Q000378:metabolism* / Q000648:ultrastructure; D002999:Clone Cells; D015536:Down-Regulation / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics*; D007150:Immunohistochemistry; D060850:LIM-Homeodomain Proteins; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D034741:RNA, Small Interfering; D018874:Spheroids, Cellular; D014157:Transcription Factors; D014404:Tubulin / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D005904:Glial Fibrillary Acidic Protein; D018398:Homeodomain Proteins; C112960:LIM homeobox protein 8; D060850:LIM-Homeodomain Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors; D014404:Tubulin; C462941:beta3 tubulin, mouse; D005680:gamma-Aminobutyric Acid; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine