Neural development of methyl-CpG-binding protein 2 null embryonic stem cells: a system for studying Rett syndrome.
Mutations in methyl-CpG-binding protein 2 (MeCP2) gene cause the neurodevelopmental disorder Rett syndrome (RTT). Here, we describe a new experimental system that efficiently elucidates the role of MeCP2 in neural development. MeCP2-null and control ES cells were generated by adenoviral conditional targeting and examined for maintenance of the undifferentiated ES cell state, neurogenesis, and gliogenesis during in vitro differentiation. In addition, dopamine release and electrophysiological features of neurons differentiated from these ES cells were examined. Loss of MeCP2 did not affect undifferentiated ES cell colony morphology and growth, or the timing or efficiency of neural stem cell differentiation into Nestin-, TuJ- or TH-positive neurons. In contrast, gliogenesis was drastically accelerated by MeCP2 deficiency. Dopamine production and release in response to a depolarizing stimulus in MeCP2-null ES-derived dopaminergic neurons was intact. However, MeCP2-null differentiated neurons showed significantly smaller voltage-dependent Na(+) currents and A-type K(+) currents, suggesting incomplete maturation. Thus, MeCP2 is not essential for maintenance of the undifferentiated ES cell state, neurogenesis, or dopaminergic function; rather, it is principally involved in inhibiting gliogenesis. Altered neuronal maturity may indirectly result from abnormal glial development and may underlie the pathogenesis of RTT. These data contribute to a better understanding of the developmental roles of MeCP2 and the pathogenesis of RTT.
Okabe
Yasunori
Y
Division of Gene Therapy and Regenerative Medicine, Kurume University, Kurume, Japan.
Kusaga
Akira
A
Takahashi
Tomoyuki
T
Mitsumasu
Chiaki
C
Murai
Yoshinaka
Y
Tanaka
Eiichiro
E
Higashi
Hideho
H
Matsuishi
Toyojiro
T
Kosai
Ken-ichiro
K
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002999:Clone Cells; D004298:Dopamine / Q000502:physiology; D055724:Electrophysiological Phenomena; D053595:Embryonic Stem Cells / Q000502:physiology*; D005822:Genetic Vectors; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D051783:Methyl-CpG-Binding Protein 2 / Q000235:genetics* / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058953:Neural Stem Cells / Q000502:physiology*; D055495:Neurogenesis / Q000502:physiology; D009457:Neuroglia / Q000502:physiology; D009474:Neurons / Q000502:physiology; D015518:Rett Syndrome / Q000235:genetics* / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005904:Glial Fibrillary Acidic Protein; D051783:Methyl-CpG-Binding Protein 2; D004298:Dopamine