Noggin and basic FGF were implicated in forebrain fate and caudal fate, respectively, of the neural tube-like structures emerging in mouse ES cell culture.
We developed neural tube-like structures accompanying neural crest-like cells by treating embryonic stem (ES) cells with retinoic acid. The structures contained pseudostratified Nestin+Vimentin+ neuroepithelial cells surrounded by Masson staining+ basement membrane. betaIIItubulin+Synaptophysin+ mature neurons and glial fibrillary acidic protein (GFAP)+ glial cells dispersed outside of the membrane. Addition of Noggin to the culture induced prominent proliferation of the neuroepithelial cells, leading to epithelial hyperstratification of the structures. mRNAs of transcription factors essential for forebrain development such as Emx1/2 and Pax6 were specifically expressed and Islet1+Lim1/2- motoneurons appeared by the addition of Noggin. In contrast, basic fibroblast growth factor (bFGF) promoted enlargement of central lumen and elongation of the structures. mRNAs of caudal markers, Gbx2, Cdx2 and Hoxb4/9 were expressed and Lim1/2+ spinal motoneurons appeared by the addition of bFGF. Addition of BMP-4 similarly brought about mild enlargement of central lumen of the structures. Interestingly, the addition of BMP-4 induced Slug+ neural crest-like cells surrounding the tube-like structures. mRNAs of Snail and dHand, other markers for neural crest cells, were also expressed by the addition of BMP-4. These results suggest that Noggin lead the neural-tube like structures to forebrain fate, whereas bFGF was involved in the caudalization. BMP-4 was implicated in emergence of the neural crest-like cells. Differentiation of ES cells by the present methods may mimic neurulation and subsequent neural development of early embryos, and elucidates the opposite effects of Noggin and bFGF for the neural tube development.
Chiba
Shunmei
S
Department of Immunology, St. Marianna University School of Medicine, 2-16-1 Sugao, 216-8511 Miyamae-ku, Kawasaki, Japan.
Kurokawa
Manae S
MS
Yoshikawa
Hideshi
H
Ikeda
Ritsuko
R
Takeno
Mitsuhiro
M
Tadokoro
Mamoru
M
Sekino
Hiroaki
H
Hashimoto
Takuo
T
Suzuki
Noboru
N
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics* / Q000494:pharmacology; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology / Q000502:physiology; D051379:Mice; D009432:Neural Crest / Q000166:cytology / Q000196:embryology / Q000502:physiology; D009924:Organ Culture Techniques; D016548:Prosencephalon / Q000166:cytology / Q000196:embryology* / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D012249:Rhombencephalon / Q000166:cytology / Q000196:embryology* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D014212:Tretinoin / Q000494:pharmacology; D000099176:Noggin Protein
D000970:Antineoplastic Agents; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D016222:Fibroblast Growth Factor 2; D012333:RNA, Messenger; D014157:Transcription Factors; D014212:Tretinoin; D000099176:Noggin Protein; C525810:Bmp4 protein, mouse