Vsx1 and Chx10 paralogs sequentially secure V2 interneuron identity during spinal cord development

Debrulle, Stéphanie; Baudouin, Christophe; Hidalgo-Figueroa, María; Pelosi, Barbara; Francius, Cédric; Rucchin, Vincent; Ronellenfitch, Kara; Chow, Robert L.; Tissir, Fadel; Lee, Soo Kyung; Clotman, Frédéric

doi:10.1007/s00018-019-03408-7

Cited by 13 publications

(11 citation statements)

References 56 publications

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“…NPCs were subsequently ventralized through the addition of the sonic hedhehog (SHH) agonist purmorphamine at a concentration of 0.5 µM, in the presence of 0.1 µM RA, resulting in the generation of MNPCs, with 74±6% of induced cells being PAX6positive + and 76±4% expressing the MNPC marker OLIG2(Briscoe et al, 2000;Jessell, 2000;Alaynick et al, 2011) at day 12 (Figure 1C and D). As expected, a small proportion of spinal ventral IN progenitors was detected in our cultures, with 1.8±0.3% of the total of differentiated cells expressing the p3 domain marker NKX2.2(Novitch et al, 2001;Sugimori et al, 2007) and 2.5±0.8% expressing the p2 domain marker CHX10(Debrulle et al, 2019;Alaynick et al, 2011) (Figure 1E and F).Continued exposure of MNPCs to RA and purmorphamine led to the generation of an enriched population of induced cells (67±7%) expressing the typical pan-MN markers ISL1…”

supporting

confidence: 83%

“…Clusters #10 and #12 were annotated as "NPCs" based on their expression of SOX1, SOX2 and MARKER OF PROLIFERATION KI-67 (MKI67) (Kan et al, 2007;Graham et al, 2003;Scholzen & Gerdes, 2000). Clusters #5 and #6 were annotated as "INs" with high expression of specific marker genes for IN progenitors such as SOX14, NKX2.2, VISUAL SYSTEM HOMEOBOX 1 (VSX1), CHX10, and SINGLE-MINDED HOMOLOG 1 (SIM1) (Ericson et al, 1997;Novitch et al, 2001;Sugimori et al, 2007;Alaynick et al 2011;Debrulle et al, 2019), as well as the mature IN marker CALBINDIN 1 (CALB1) (Alvarez et al, 2005). Cluster #2 was annotated as "astrocytic glial cells" based on high expression of S100 CALCIUM BINDING PROTEIN B…”

Section: Characterization Of Cellular Heterogeneity Of Human Ipsc-dermentioning

confidence: 99%

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Characterization of human-iPSCs derived spinal motor neurons by single-cell RNA sequencing

Thiry

Hamel

Pluchino

et al. 2019

Preprint

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AbstractHuman induced pluripotent stem cells (iPSCs) offer the opportunity to generate specific cell types from healthy and diseased individuals, allowing the study of mechanisms of early human development, modelling a variety of human diseases, and facilitating the development of new therapeutics. Human iPSC-based applications are often limited by the variability among iPSC lines originating from a single donor, as well as the heterogeneity among specific cell types that can be derived from iPSCs. The ability to deeply phenotype different iPSC-derived cell types is therefore of primary importance to the successful and informative application of this technology. Here we describe a combination of motor neuron (MN) derivation and single-cell RNA sequencing approaches to generate and characterize specific MN subtypes obtained from human iPSCs. Our studies provide evidence for rapid and robust generation of MN progenitor cells that can give rise to a heterogenous population of brainstem and spinal cord MNs. Approximately 58% of human iPSC-derived MNs display molecular characteristics of lateral motor column MNs, ∼19% of induced MNs resemble hypaxial motor column MNs, while ∼6% of induced MNs have features of medial motor column MNs. The present study has the potential to improve our understanding of iPSC-derived MN subtype function and dysfunction, possibly leading to improved iPSC-based applications for the study of human MN biology and diseases.

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supporting

confidence: 83%

Section: Characterization Of Cellular Heterogeneity Of Human Ipsc-dermentioning

confidence: 99%

Characterization of human-iPSCs derived spinal motor neurons by single-cell RNA sequencing

Thiry

Hamel

Pluchino

et al. 2019

Preprint

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“…Since Vsx1 is only transiently maintained in V2a sub-lineage at early specification stage, the subsequent steady expression of Vsx2 may ensure the persistent inhibition of Tal1 erroneous expression in V2a interneuron sub-lineage. A recent study in mice has observed that Vsx1 and Vsx2 act successively to secure V2 interneuron identity and Vsx2 can inhibit Gata3 expression and V2b interneuron differentiation [36]. We have reason to believe that, in zebrafish, the transient maintenance of Vsx1 in V2a cells inhibits Tal1 expression and stimulates Vsx2 expression at the onset of V2a and V2b diversification, resulting in adequate segregation of Vsx2 and Tal1 expression and proper V2a vs. V2b fate decision.…”

Section: Direct Repression Of Tal1 By Paired-like Homeodomain Repressmentioning

confidence: 99%

Zygotic Vsx1 Plays a Key Role in Defining V2a Interneuron Sub-Lineage by Directly Repressing tal1 Transcription in Zebrafish

Zhang

Zhao

et al. 2020

IJMS

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In the spinal cord, excitatory V2a and inhibitory V2b interneurons are produced together by the final division of common P2 progenitors. During V2a and V2b diversification, Tal1 is necessary and sufficient to promote V2b differentiation and Vsx2 suppresses the expression of motor neuron genes to consolidate V2a interneuron identity. The expression program of Tal1 is triggered by a Foxn4-driven regulatory network in the common P2 progenitors. Why the expression of Tal1 is inhibited in V2a interneurons at the onset of V2a and V2b sub-lineage diversification remains unclear. Since transcription repressor Vsx1 is expressed in the P2 progenitors and newborn V2a cells in zebrafish, we investigated the role of Vsx1 in V2a fate specification during V2a and V2b interneuron diversification in this species by loss and gain-of-function experiments. In vsx1 knockdown embryos or knockout Go chimeric embryos, tal1 was ectopically expressed in the presumptive V2a cells, while the generation of V2a interneurons was significantly suppressed. By contrast, in vsx1 overexpression embryos, normal expression of tal1 in the presumptive V2b cells was suppressed, while the generation of V2a interneuron was expanded. Chromatin immunoprecipitation and electrophoretic mobility shift assays in combination with core consensus sequence mutation analysis further revealed that Vsx1 can directly bind to tal1 promoter and repress tal1 transcription. These results indicate that Vsx1 can directly repress tal1 transcription and plays an essential role in defining V2a interneuron sub-lineage during V2a and V2b sub-lineage diversification in zebrafish.

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“…For instance, the homeodomain transcription factor Chx10/Vsx2 is expressed and is required for the V2a interneuron. In the absence of Chx10, progenitor cells acquire different neuronal identities of the motor neuron (MN), which is located in the adjacent domain of the neural tube [11,12]. As such, there are many transcription factors that regulate neuronal differentiation and/or acquisition of specific functions.…”

Section: Introductionmentioning

confidence: 99%

Sox14 is essential for initiation of interneuron differentiation in the chick spinal cord

Katsuyama

Kadoya

Shirai

et al. 2021

Preprint

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The neural tube comprises several different types of progenitors and postmitotic neurons that coordinately act with each other to play integrated functions. Its development consists of two phases: proliferation of progenitor cells and differentiation into postmitotic neurons. How progenitor cells differentiate into each corresponding neuron is an important question for understanding the mechanisms of neuronal development. Here we introduce one of the Sox transcription factors, Sox14, which plays an essential role in the promotion of neuronal differentiation. Sox14 belongs to the SoxB subclass and its expression starts in the progenitor regions before neuronal differentiation is initiated at the trunk level of the neural tube. After neuronal differentiation is initiated, Sox14 expression gradually becomes confined to the V2a region of the neural tube, where Chx10 is co-expressed. Overexpression of Sox14 restricts progenitor cell proliferation. Conversely, the blockade of Sox14 expression by the RNAi strategy inhibits V2a neuron differentiation and causes expansion of the progenitor domain. We further found that Sox14 acted as a transcriptional activator. Taken together, Sox14 acts as a modulator of cell proliferation and an initiator protein for neuronal differentiation in the intermediate region of the neural tube.

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Vsx1 and Chx10 paralogs sequentially secure V2 interneuron identity during spinal cord development

Cited by 13 publications

References 56 publications

Characterization of human-iPSCs derived spinal motor neurons by single-cell RNA sequencing

Characterization of human-iPSCs derived spinal motor neurons by single-cell RNA sequencing

Zygotic Vsx1 Plays a Key Role in Defining V2a Interneuron Sub-Lineage by Directly Repressing tal1 Transcription in Zebrafish

Sox14 is essential for initiation of interneuron differentiation in the chick spinal cord

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