ABSTRACT
Members of the T-box transcription factor family (Tbx) are associated with several human syndromes during embryogenesis. Nevertheless, their functions within the developing CNS remain poorly characterized. Tbx20 is expressed by migrating branchiomotor/visceromotor (BM/VM) neurons within the hindbrain during neuronal circuit formation. We examined Tbx20 function in BM/VM cells using conditional Tbx20-null mutant mice to delete the gene in neurons. Hindbrain rhombomere patterning and the initial generation of post-mitotic BM/VM neurons were normal in Tbx20 mutants. However, Tbx20 was required for the tangential (caudal) migration of facial neurons, the lateral migration of trigeminal cells and the trans-median movement of vestibuloacoustic neurons. Facial cell soma migration defects were associated with the coordinate downregulation of multiple components of the planar cell polarity pathway including Fzd7, Wnt11, Prickle1, Vang1 and Vang2. Our study suggests that Tbx20 programs a variety of hindbrain motor neurons for migration, independent of directionality, and in facial neurons is a positive regulator of the non-canonical Wnt signaling pathway.
Subject(s)
Cranial Nerves/embryology , Motor Neurons/cytology , T-Box Domain Proteins/genetics , T-Box Domain Proteins/metabolism , Animals , Axons/metabolism , Cell Movement , Cell Polarity , Cranial Nerves/cytology , Embryonic Development/genetics , Facial Nerve/cytology , Facial Nerve/embryology , Gene Expression Profiling , Mice , Mice, Transgenic , Motor Neurons/metabolism , Mutation , Signal Transduction , Trigeminal Nerve/cytology , Trigeminal Nerve/embryology , Vestibulocochlear Nerve/cytology , Vestibulocochlear Nerve/embryology , Wnt Proteins/metabolismABSTRACT
Neuronal gene transcription is repressed in non-neuronal cells by the repressor element 1 (RE-1)-silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) complex. To understand how this silencing is achieved, we examined a family of class-C RNA polymerase II (RNAPII) carboxyl-terminal domain (CTD) phosphatases [small CTD phosphatases (SCPs) 1 to 3], whose expression is restricted to non-neuronal tissues. We show that REST/NRSF recruits SCPs to neuronal genes that contain RE-1 elements, leading to neuronal gene silencing in non-neuronal cells. Phosphatase-inactive forms of SCP interfere with REST/NRSF function and promote neuronal differentiation of P19 stem cells. Likewise, small interfering RNA directed to the single Drosophila SCP unmasks neuronal gene expression in S2 cells. Thus, SCP activity is an evolutionarily conserved transcriptional regulator that acts globally to silence neuronal genes.
Subject(s)
Gene Silencing , Neurons/physiology , Phosphoprotein Phosphatases/metabolism , Repressor Proteins/metabolism , Transcription Factors/metabolism , Animals , Basic Helix-Loop-Helix Transcription Factors , Cell Differentiation , Cell Line , Chromatin Immunoprecipitation , DNA-Binding Proteins/metabolism , Down-Regulation , Drosophila/genetics , Drosophila/metabolism , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Gene Expression Profiling , Gene Expression Regulation , Humans , In Situ Hybridization , Mice , Nerve Tissue Proteins/metabolism , Neurons/cytology , Nuclear Proteins , Phosphoprotein Phosphatases/genetics , Phosphorylation , RNA Interference , Regulatory Sequences, Nucleic Acid , TCF Transcription Factors , Transcription Factor 7-Like 1 Protein , Tretinoin/pharmacologyABSTRACT
Spinal motor neurons and oligodendrocytes are generated sequentially from a common pool of progenitors termed pMN cells. Olig2 is a bHLH-class transcription factor in pMN cells, but it has remained unclear how its transcriptional activity is modulated to first produce motor neurons and then oligodendrocytes. Previous studies have shown that Olig2 primes pMN cells to become motor neurons by triggering the expression of Ngn2 and Lhx3. Here we show that Olig2 also antagonizes the premature expression of post-mitotic motor neuron genes in pMN cells. This blockade is counteracted by Ngn2, which accumulates heterogeneously in pMN cells, thereby releasing a subset of the progenitors to differentiate and activate expression of post-mitotic motor neuron genes. The antagonistic relationship between Ngn2 and Olig2 is mediated by protein interactions that squelch activity as well as competition for shared DNA-binding sites. Our data support a model in which the Olig2/Ngn2 ratio in progenitor cells serves as a gate for timing proper gene expression during the development of pMN cells: Olig2(high) maintains the pMN state, thereby holding cells in reserve for oligodendrocyte generation, whereas Ngn2(high) favors the conversion of pMN cells into post-mitotic motor neurons.
Subject(s)
Cell Differentiation/genetics , Gene Expression Regulation, Developmental/genetics , Homeodomain Proteins/biosynthesis , Motor Neurons/metabolism , Nerve Tissue Proteins/biosynthesis , Oligodendroglia/metabolism , Stem Cells/metabolism , Animals , Basic Helix-Loop-Helix Transcription Factors , Cell Differentiation/physiology , Chick Embryo , Embryonic Development/genetics , Embryonic Development/physiology , Gene Expression Regulation, Developmental/physiology , Homeodomain Proteins/genetics , LIM-Homeodomain Proteins , Mice , Motor Neurons/cytology , Nerve Tissue Proteins/genetics , Oligodendrocyte Transcription Factor 2 , Oligodendroglia/cytology , Transcription FactorsABSTRACT
The underlying transcriptional mechanisms that establish the proper spatial and temporal pattern of gene expression required for specifying neuronal fate are poorly defined. We have characterized how the Hb9 gene is expressed in developing motoneurons in order to understand how transcription is directed to specific cells within the developing CNS. We found that non-specific general-activator proteins such as E2F and Sp1 are capable of driving widespread low level transcription of Hb9 in many cell types throughout the neural tube; however, their activity is modulated by specific repressor and activator complexes. The general-activators of Hb9 are suppressed from triggering inappropriate transcription by repressor proteins Irx3 and Nkx2.2. High level motoneuron expression is achieved by assembling an enhancesome on a compact evolutionarily-conserved segment of Hb9 located from -7096 to -6896. The ensemble of LIM-HD and bHLH proteins that interact with this enhancer change as motoneuron development progresses, facilitating both the activation and maintenance of Hb9 expression in developing and mature motoneurons. These findings provide direct support for the derepression model of gene regulation and cell fate specification in the neural tube, as well as establishing a role for enhancers in targeting gene expression to a single neuronal subtype in the spinal cord.
