Abstract
MicroRNAs have emerged as key posttranscriptional regulators of gene expression during vertebrate development. We show that the miR-200 family plays a crucial role for the proper generation and survival of ventral neuronal populations in the murine midbrain/hindbrain region, including midbrain dopaminergic neurons, by directly targeting the pluripotency factor Sox2 and the cell-cycle regulator E2F3 in neural stem/progenitor cells. The lack of a negative regulation of Sox2 and E2F3 by miR-200 in conditional Dicer1 mutants (En1(+/Cre); Dicer1(flox/flox) mice) and after miR-200 knockdown in vitro leads to a strongly reduced cell-cycle exit and neuronal differentiation of ventral midbrain/hindbrain (vMH) neural progenitors, whereas the opposite effect is seen after miR-200 overexpression in primary vMH cells. Expression of miR-200 is in turn directly regulated by Sox2 and E2F3, thereby establishing a unilateral negative feedback loop required for the cell-cycle exit and neuronal differentiation of neural stem/progenitor cells. Our findings suggest that the posttranscriptional regulation of Sox2 and E2F3 by miR-200 family members might be a general mechanism to control the transition from a pluripotent/multipotent stem/progenitor cell to a postmitotic and more differentiated cell.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Age Factors
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Animals
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Cell Count
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Cell Cycle / genetics
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Cell Cycle / physiology*
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Cell Death / genetics
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Cell Differentiation / genetics
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Cell Differentiation / physiology*
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Cells, Cultured
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Chlorocebus aethiops
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DEAD-box RNA Helicases / genetics
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DEAD-box RNA Helicases / metabolism
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Deoxyuridine / analogs & derivatives
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Deoxyuridine / metabolism
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E2F3 Transcription Factor / genetics
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E2F3 Transcription Factor / metabolism*
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Embryo, Mammalian
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Female
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Gene Expression Regulation, Developmental / genetics
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Gene Expression Regulation, Developmental / physiology*
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Green Fluorescent Proteins / genetics
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Homeodomain Proteins / genetics
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Homeodomain Proteins / metabolism
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Humans
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Male
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Mesencephalon / cytology
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Mice
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Mice, Inbred C57BL
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Mice, Transgenic
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MicroRNAs / genetics
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MicroRNAs / metabolism*
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Models, Biological
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Mutation / genetics
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Nerve Tissue Proteins / metabolism
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Neural Stem Cells / physiology*
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Rhombencephalon / cytology
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Ribonuclease III / genetics
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Ribonuclease III / metabolism
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SOXB1 Transcription Factors / genetics
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SOXB1 Transcription Factors / metabolism*
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Serotonin / metabolism
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Signal Transduction / genetics
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Transcription Factor Brn-3A / metabolism
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Transcription Factors / metabolism
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Transfection
Substances
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E2F3 Transcription Factor
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E2f3 protein, mouse
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En1 protein, mouse
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Homeodomain Proteins
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MicroRNAs
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Mirn200 microRNA, mouse
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Nerve Tissue Proteins
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Pou4f1 protein, mouse
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SOXB1 Transcription Factors
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Sox2 protein, mouse
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Transcription Factor Brn-3A
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Transcription Factors
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enhanced green fluorescent protein
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homeobox protein PITX3
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Green Fluorescent Proteins
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Serotonin
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Dicer1 protein, mouse
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Ribonuclease III
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DEAD-box RNA Helicases
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5-ethynyl-2'-deoxyuridine
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Deoxyuridine