TY - JOUR
T1 - Distinct effects of caudalizing factors on regional specification of embryonic stem cell-derived neural precursors
AU - Irioka, Takashi
AU - Watanabe, Kiichi
AU - Mizusawa, Hidehiro
AU - Mizuseki, Kenji
AU - Sasai, Yoshiki
N1 - Funding Information:
We are grateful to the members of the Sasai lab for comments on this work and to Ayaka Nishiyama for technical assistance on cell culture. This work was supported by grants-in-aid (YS) from MEXT, the Kobe Cluster Project and the Leading Project.
PY - 2005/1/1
Y1 - 2005/1/1
N2 - Recent embryological studies have implicated several "caudalizing factors" in the caudal specification of the central nervous system (CNS). In this study, we have examined the effects of three candidate caudalizing factors on neural precursors induced from embryonic stem (ES) cells by the stromal cell-derived inducing activity (SDIA) method. Among retinoic acid (RA), Wnt and FGF signals, RA causes the strongest level of caudalization: inducing suppression of forebrain differentiation and promotion of caudal CNS specification. Obvious suppression of the telencephalic marker Bf1 and that of the forebrain marker Otx2 occur at 2×10 -8 and 2×10 -7 M, respectively. Activation of the caudal marker genes such as Hoxb9 is observed in a dose-dependent manner over the range of 2×10 -9-2×10 -6 M. Suppression of the forebrain genes has a narrow critical period of RA response during the early culture phase. In contrast, significant induction of the caudal genes is evoked by a 1-day exposure to RA at any time between days 3 and 8. RA treatment not only induces caudal specification but also inhibits differentiation of ventral CNS tissues, particularly of floor plate cells. FGF4 induces partial caudalization while Wnt-3A exhibits weak caudalizing activities only in the presence of RA. These findings provide useful information on the proper selection of combination of signaling molecules, doses and timing for steering ES cell differentiation by caudalizing factors into caudal neural fates.
AB - Recent embryological studies have implicated several "caudalizing factors" in the caudal specification of the central nervous system (CNS). In this study, we have examined the effects of three candidate caudalizing factors on neural precursors induced from embryonic stem (ES) cells by the stromal cell-derived inducing activity (SDIA) method. Among retinoic acid (RA), Wnt and FGF signals, RA causes the strongest level of caudalization: inducing suppression of forebrain differentiation and promotion of caudal CNS specification. Obvious suppression of the telencephalic marker Bf1 and that of the forebrain marker Otx2 occur at 2×10 -8 and 2×10 -7 M, respectively. Activation of the caudal marker genes such as Hoxb9 is observed in a dose-dependent manner over the range of 2×10 -9-2×10 -6 M. Suppression of the forebrain genes has a narrow critical period of RA response during the early culture phase. In contrast, significant induction of the caudal genes is evoked by a 1-day exposure to RA at any time between days 3 and 8. RA treatment not only induces caudal specification but also inhibits differentiation of ventral CNS tissues, particularly of floor plate cells. FGF4 induces partial caudalization while Wnt-3A exhibits weak caudalizing activities only in the presence of RA. These findings provide useful information on the proper selection of combination of signaling molecules, doses and timing for steering ES cell differentiation by caudalizing factors into caudal neural fates.
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U2 - 10.1016/j.devbrainres.2004.10.004
DO - 10.1016/j.devbrainres.2004.10.004
M3 - Article
C2 - 15617756
AN - SCOPUS:11144332448
SN - 0165-3806
VL - 154
SP - 63
EP - 70
JO - Developmental Brain Research
JF - Developmental Brain Research
IS - 1
ER -