Armisén R et al. (2002), Repressor element-1 silencing transcription/neu...

XB-ART-6475

J Neurosci 2002 Oct 01;2219:8347-51.

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Repressor element-1 silencing transcription/neuron-restrictive silencer factor is required for neural sodium channel expression during development of Xenopus.

Armisén R , Fuentes R , Olguín P , Cabrejos ME , Kukuljan M .

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The ability of neurons to fire rapid action potential relies on the expression of voltage-gated sodium channels; the onset of the transcription of genes that encode these channels occurs during early neuronal development. The factors that direct and regulate the specific expression of ion channels are not well understood. Repressor element-1 silencing transcription/neuron-restrictive silencer factor (REST/NRSF) is a transcriptional regulator characterized as a repressor of the expression of NaV1.2, the gene encoding the voltage-gated sodium channel most abundantly expressed in the CNS, as well as of the expression of numerous other neuronal genes. In mammals, REST/NRSF is expressed mostly in non-neural cell types and immature neurons, and it is downregulated on neural maturation. To understand the mechanisms that govern sodium channel gene transcription and to explore the role of REST/NRSF in vivo, we inhibited REST/NRSF action in developing Xenopus laevis embryos by means of a dominant negative protein or antisense oligonucleotides. Contrary to what was expected, these maneuvers result in the decrease of the expression of the NaV1.2 gene, as well as of other neuronal genes in the primary spinal neurons and cranial ganglia, without overt perturbation of neurogenesis. These results, together with the demonstration of robust REST/NRSF expression in primary spinal neurons, suggest that REST/NRSF is required for the acquisition of the differentiated functional neuronal phenotype during early development. Furthermore, they suggest that REST/NRSF may be used to activate or repress transcription of neuronal genes in distinct cellular and developmental contexts.

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Species referenced: Xenopus laevis
Genes referenced: myc nav1 pax3 rest scn1a snai2 sox2 stmn2 tlx3 tubb2b
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	Fig. 1. NaV1.2 and REST/NRSF are expressed in neural tissues during development of X. laevis.a–d, ISH for xNaV1.2: expression is restricted to primary spinal neurons (sn), cranial ganglia primordia (cg), and olfactory placodes (op). In a section of a stage 18 embryo, xNaV 1.2 expression is observed in the lateral and ventral regions of the neural tube. a, Dorsal view, stage 18; b, lateral view, stage 18;c, anterior view, stage 24; d, central transversal section, stage 18; n, notochord;m, presomitic mesoderm; e,left, ISH showing diffuse expression of REST during neurulation (stage 18), including neural folds; right, a sense REST probe does not produce significant labeling in a stage 18 embryo; f, at stage 35, REST/NRSF expression is stronger in the anterior neural tissue; g, RT-PCR showing the coexpression of xNaV1.2 and REST/NRSF in dissected tissues at the stages annotated; np, neural plate; nt, neural tube; pm, presomitic mesoderm; s, somites. The constitutively expressed transcript EF1α is shown as a control.
	Fig. 2. DBD-xREST binds specifically to the RE-1 and inhibits xNaV1.2 expression. a, Electrophoretical mobility shift assay analysis of the complex formation by the use of protein extracts of noninjected embryos (lanes 1–2) and embryos injected with the DBD-xREST RNA (lanes 3–9). Two or 10 μg of total protein was used in each assay. DBD-xREST binds to RE-1 (lanes 3–6). The addition of antibody against the Myc epitope produces the super-retardation of the complex (lanes 7–9). The binding specificity of the complex was evaluated by the addition of 2- or 50-fold excess of cold probe (lanes 5–6, 8–9). The probe alone is shown inLane 10. Arrows indicate the complexes.b, Expression of xREST-DBD inhibits xNaV1.2 expression during neurulation. ISH for xNaV1.2; the injected side is marked by anasterisk. Notice a single line of xNaV1.2 labeling, in contrast with Figure 1 a. c, Injection of a morpholino-modified antisense oligonucleotide targeting xREST/NRSF inhibits xNaV 1.2 expression at the time of neurulation.d, Detail of a noninjected embryo (top panel) and embryos injected with xREST-DBD RNA (middle panel) or the antisense oligonucleotide (bottom panel) in one of two cells.
	Fig. 3. xREST-DBD decreases the expression of other neuronal genes but does not perturb neural induction.a–c, ISH for N-tubulin; the injected side is marked by an asterisk; b, both lateral views;c, anterior view of the same embryo, where thearrowheads signal the label of the primary spinal neurons (sn) and cranial ganglia (cg).d, xREST-DBD represses SCG10 expression.Double arrowheads signal the expected normal position of N-tubulin or SCG10 expression. The spotted labeling in the injected side (a) and thediffuse hue in c result from β-galactosidase activity staining. e–h, xREST-DBD does not affect neural induction and early differentiation. Expression patterns of Sox2 (e), Slug (f), Hox11L12 (g), and Pax-3 (h). e, h, Anterior views; f, g, dorsal views.
	scn1a (sodium channel, voltage gated, type I alpha subunit) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 18, lateral view, anterior right, dorsal up.
	scn1a (sodium channel, voltage gated, type I alpha subunit) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 24, anterior view, dorsal up. Key: cg= cranial/trigeminal ganglion, op: olfactory placode.
	rest (RE1-silencing transcription factor) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 18, dorsal view, anterior up.
	rest (RE1-silencing transcription factor) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35, lateral view, anterior right, dorsal up.
	tlx3 (T cell leukemia homeobox 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 18, dorsal view, anterior up.

References [+] :

Andrés, CoREST: a functional corepressor required for regulation of neural-specific gene expression. 1999, Pubmed

Andrés, CoREST: a functional corepressor required for regulation of neural-specific gene expression. 1999, Pubmed
Ballas, Regulation of neuronal traits by a novel transcriptional complex. 2001, Pubmed
Bessis, The neuron-restrictive silencer element: a dual enhancer/silencer crucial for patterned expression of a nicotinic receptor gene in the brain. 1997, Pubmed
Burger, Xenopus spinal neurons express Kv2 potassium channel transcripts during embryonic development. 1996, Pubmed , Xenbase
Chen, NRSF/REST is required in vivo for repression of multiple neuronal target genes during embryogenesis. 1998, Pubmed
Chong, REST: a mammalian silencer protein that restricts sodium channel gene expression to neurons. 1995, Pubmed
Desarmenien, In vivo development of voltage-dependent ionic currents in embryonic Xenopus spinal neurons. 1993, Pubmed , Xenbase
Grimes, The co-repressor mSin3A is a functional component of the REST-CoREST repressor complex. 2000, Pubmed
Gurantz, Temporal regulation of Shaker- and Shab-like potassium channel gene expression in single embryonic spinal neurons during K+ current development. 1996, Pubmed , Xenbase
Hartenstein, Early neurogenesis in Xenopus: the spatio-temporal pattern of proliferation and cell lineages in the embryonic spinal cord. 1989, Pubmed , Xenbase
Huang, Transcriptional repression by REST: recruitment of Sin3A and histone deacetylase to neuronal genes. 1999, Pubmed
Jepsen, Combinatorial roles of the nuclear receptor corepressor in transcription and development. 2000, Pubmed
Kallunki, The neural restrictive silencer element can act as both a repressor and enhancer of L1 cell adhesion molecule gene expression during postnatal development. 1998, Pubmed
Kraner, Silencing the type II sodium channel gene: a model for neural-specific gene regulation. 1992, Pubmed
Kuwahara, The neuron-restrictive silencer element-neuron-restrictive silencer factor system regulates basal and endothelin 1-inducible atrial natriuretic peptide gene expression in ventricular myocytes. 2001, Pubmed
Mori, A common silencer element in the SCG10 and type II Na+ channel genes binds a factor present in nonneuronal cells but not in neuronal cells. 1992, Pubmed
Naruse, Neural restrictive silencer factor recruits mSin3 and histone deacetylase complex to repress neuron-specific target genes. 1999, Pubmed
O'Dowd, Development of voltage-dependent calcium, sodium, and potassium currents in Xenopus spinal neurons. 1988, Pubmed , Xenbase
Olson, Onset of electrical excitability during a period of circus plasma membrane movements in differentiating Xenopus neurons. 1996, Pubmed , Xenbase
Palm, Neuronal expression of zinc finger transcription factor REST/NRSF/XBR gene. 1998, Pubmed , Xenbase
Paquette, Constitutive expression of the neuron-restrictive silencer factor (NRSF)/REST in differentiating neurons disrupts neuronal gene expression and causes axon pathfinding errors in vivo. 2000, Pubmed
Rao, New functions for DNA binding domains. 2001, Pubmed
Ribera, A potassium channel gene is expressed at neural induction. 1990, Pubmed , Xenbase
Ribera, A critical period of transcription required for differentiation of the action potential of spinal neurons. 1989, Pubmed , Xenbase
Schoenherr, The neuron-restrictive silencer factor (NRSF): a coordinate repressor of multiple neuron-specific genes. 1995, Pubmed
Schoenherr, Identification of potential target genes for the neuron-restrictive silencer factor. 1996, Pubmed
Scholl, A zinc finger protein that represses transcription of the human MHC class II gene, DPA. 1996, Pubmed
Seth, Repressor element silencing transcription factor/neuron-restrictive silencing factor (REST/NRSF) can act as an enhancer as well as a repressor of corticotropin-releasing hormone gene transcription. 2001, Pubmed
Spitzer, The development of the action potential mechanism of amphibian neurons isolated in culture. 1976, Pubmed , Xenbase
Vincent, Antisense suppression of potassium channel expression demonstrates its role in maturation of the action potential. 2000, Pubmed , Xenbase
de la Calle-Mustienes, The Xiro-repressed gene CoREST is expressed in Xenopus neural territories. 2002, Pubmed , Xenbase