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The oncoprotein v-Myb activates transcription of Gremlin 2 during differentiation of the chicken neural crest to melanoblasts

ABSTRACT: The neural crest (NC) is a transient dynamic structure of ectodermal origin, found in early vertebrate embryos. The multipotential NC cells migrate along well defined routes, differentiate to various cells types including melanocytes and participate in the formation of various permanent tissues. Abnormal development of NC cells causes several human diseases – neurocristopathies. As there is only limited information about the molecular mechanisms controlling early events in melanocyte specification and development, we exploited the AMV v-Myb transcriptional regulator, which directs differentiation of in vitro chicken NC cells to the melanocyte lineage. This activity is strictly dependent on v-Myb specifically binding to the Myb recognition DNA element (MRE). The two tamoxifen-inducible v-myb alleles were constructed, one which recognizes the MRE and one which does not. These were activated in ex-ovo NC cells, and the expression profiles of resulting cells were analyzed using Affymetrix microarrays and RT-PCR. These approaches revealed up-regulation of the BMP antagonist gremlin 2 mRNA, and down-regulation of mRNAs encoding several epithelial genes including KRT19 as very early events following the activation of melanocyte differentiation by v-Myb.

ORGANISM(S): Gallus gallus

PROVIDER: GSE39346 | GEO | 2012/07/14

SECONDARY ACCESSION(S): PRJNA170678

REPOSITORIES: GEO

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The oncoprotein v-Myb activates transcription of Gremlin 2 during differentiation of the chicken neural crest to melanoblasts

Project description:The neural crest (NC) is a transient dynamic structure of ectodermal origin, found in early vertebrate embryos. The multipotential NC cells migrate along well defined routes, differentiate to various cells types including melanocytes and participate in the formation of various permanent tissues. Abnormal development of NC cells causes several human diseases M-bM-^@M-^S neurocristopathies. As there is only limited information about the molecular mechanisms controlling early events in melanocyte specification and development, we exploited the AMV v-Myb transcriptional regulator, which directs differentiation of in vitro chicken NC cells to the melanocyte lineage. This activity is strictly dependent on v-Myb specifically binding to the Myb recognition DNA element (MRE). The two tamoxifen-inducible v-myb alleles were constructed, one which recognizes the MRE and one which does not. These were activated in ex-ovo NC cells, and the expression profiles of resulting cells were analyzed using Affymetrix microarrays and RT-PCR. These approaches revealed up-regulation of the BMP antagonist gremlin 2 mRNA, and down-regulation of mRNAs encoding several epithelial genes including KRT19 as very early events following the activation of melanocyte differentiation by v-Myb. Comparison of gene expression profiles of chicken neural crest cells constitutively expressing 4-OH-tamoxifen inducible v-myb with mutated leucine zipper region or a version with an additional point mutation (N118D) in the DNA-binding domain. Three biological replicates were analyzed for each group.

2012-07-13 | E-GEOD-39346 | biostudies-arrayexpress

Murine lung tissue: Gremlin-1 transgenic vs. wild type

Project description:Transcriptional profiling of murine lung tissue comparing wild type to gremlin-1 transgenic untreated or silicon dioxide (silica)-treated for two months. Gremlin-1 transgene expression in lung type II epithelial cells under SPC promoter. Aim was to compare expression of fibrosis and inflammation associated genes.

2016-12-21 | GSE80406 | GEO

The effect of anti-Gremlin-1 treatment on human cirrhotic precision-cut liver slices

Project description:Gremlin-1 has been implicated in liver fibrosis in metabolic dysfunction-associated steatohepatitis (MASH) via inhibition of bone-morphogenetic protein (BMP) signalling and has thereby been identified as a potential therapeutic target. Using human cirrhotic precision-cut liver slices (PCLS) as an ex vivo model systems of MASH fibrosis, we show that neutralisation of Gremlin-1 activity with monoclonal therapeutic antibodies does not reduce liver inflammation or liver fibrosis. Overall, our findings suggest a redundant role for Gremlin-1 in the pathogenesis of liver fibrosis, which is unamenable to therapeutic targeting.

2024-02-21 | GSE245977 | GEO

FSH-deficiency in gonadotrope-specific GATA2 knockout mice is associated with a loss of gremlin expression [snATAC-Seq]

Project description:Mammalian reproduction is dependent on the gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone, which are secreted by pituitary gonadotrope cells. The zinc-finger transcription factor, GATA2, was previously implicated in FSH production in male mice, although its mechanisms of action and role in females were not determined. To address these gaps in knowledge, we generated and analyzed gonadotrope-specific Gata2 knockout mice using the Cre-lox system. While conditional knockout (cKO) males exhibited ~50% reductions in serum FSH levels and pituitary FSHβ subunit (Fshb) expression relative to controls, FSH production was apparently normal in cKO females. RNA-seq analysis of purified gonadotropes from control and cKO males revealed a profound decrease in expression of gremlin (Grem1), a bone morphogenetic protein (BMP) antagonist. Grem1 was expressed in gonadotropes, but not other cell lineages, in the adult male mouse pituitary. Gata2, Grem1, and Fshb mRNA levels were significantly higher in pituitaries of wild-type males relative to females but decreased in males treated with estradiol and increased following ovariectomy in control but not cKO females. Recombinant gremlin stimulated Fshb expression in pituitary cultures from wild-type mice. Collectively, the data suggest that GATA2 promotes Grem1 expression in gonadotropes and that the gremlin protein potentiates FSH production. The mechanisms of gremlin action have not yet been established but may involve attenuation of BMP binding to activin type II receptors in gonadotropes, potentiating induction of Fshb transcription by activins or related ligands.

2022-07-14 | GSE190064 | GEO

FSH-deficiency in gonadotrope-specific GATA2 knockout mice is associated with a loss of gremlin expression [snRNA-Seq]

2022-07-14 | GSE190060 | GEO

Comparison of expression data between control and Ovo1 morphant zebrafish embryos

Project description:A fundamental issue in cell biology is how migratory cell behaviors are controlled by dynamically regulated cell adhesion. Vertebrate neural crest (NC) cells rapidly alter cadherin expression and localization at the cell surface during migration. Secreted Wnts induce some of these changes in NC adhesion and also promote specification of NC-derived pigment cells. Here, we show that the zebrafish transcription factor Ovo1 is a Wnt target gene that controls migration of pigment precursors by regulating the intracellular movements of N-cadherin (Ncad). Ovo1 genetically interacts with Ncad and its depletion causes Ncad to accumulate inside cells. Ovo1-deficient embryos strongly upregulate factors involved in intracellular trafficking, including several rab GTPases, known to modulate cellular localization of cadherins. Surprisingly, NC cells express high levels of many of these rab genes in the early embryo, chemical inhibitors of Rab functions rescue NC development in Ovo1-deficient embryos and overexpression of a Rab-interacting protein leads to similar defects in NC migration. These results suggest that Ovo proteins link Wnt signaling to intracellular trafficking pathways that localize Ncad in NC cells and allow them to migrate. Similar processes probably occur in other cell types in which Wnt signaling promotes migration.

2010-04-28 | GSE21539 | GEO

Gene expression profiling analysis of murine Villin-Gremlin villi compared to age-matched control mice

Project description:This study was designed to assess the global transcriptional differences between villin-Grem villi compared to normal villi of age-matched mice. We found changes in key gene signatures asscoiated with tumorigenesis. Total RNA obtained from isolated villi from Villin-Gremlin mouse (n=6) and wildtype controls (n=6)

2014-10-24 | E-GEOD-62307 | biostudies-arrayexpress

Comparison of expression data between control and Ovo1 morphant zebrafish embryos

2010-04-27 | E-GEOD-21539 | biostudies-arrayexpress

Generation of neural crest cells by direct lineage conversion

Project description:Neural crest cells (NC cells) are multipotent cells that emerge from the dorsal region of a fusing neural tube and migrate throughout the developing embryo. Due to their multipotency and divergent roles during embryogenesis, the generation and differentiation of NC cells should be elucidated in more detail. However, difficulties have been associated with isolating and manipulating NC cells for investigation because a limited number of NC cells are generated in a short period of embryogenesis. To overcome this issue, we attempted to directly convert mouse fibroblasts into NC cells using transcription factors specifically expressed in NC cells. We analyzed the molecular profiles of NC cells and neural tube cells in 9.5-day mouse embryos, and identified the transcription factors that were strongly expressed in NC cells. Among these transcription factors, Sox10 together with c-Myc and Klf4 directly converted mouse fibroblasts into NC cells. The converted NC cells differentiated into neurons, glial cells, smooth muscle cells, adipocytes, and osteoblasts, exhibiting differentiation abilities similar to those of in vivo NC cells. They also displayed a self-renewal ability while maintaining their differentiation ability. In in ovo transplantation, the converted NC cells migrated along NC cell migration pathways. Therefore, directly converted NC cells can contribute to the progression of NC cell studies.

2017-02-10 | GSE63896 | GEO

The Ednrb–Aim2–AKT axis regulates neural crest-derived melanoblast proliferation during early development

Project description:Ednrb is specifically required to develop neural crest (NC) stem cell-derived lineages. However, it is still unknown why Ednrb signaling is only needed for the early development of melanoblasts in the skin and eye. We show that Ednrb is required for the proliferation of melanoblasts during early mouse development. To understand the mechanism of melanoblast proliferation, we found that the gene absent in melanoma 2 (Aim2) is upregulated in Ednrb-deficient NC cells by RNA-sequencing analysis. Consequently, the knockdown or knockout of Aim2 partially rescued the proliferation of Ednrb-deficient melanoblasts. Conversely, the overexpression of Aim2 in melanoblasts suppressed their proliferation. We further show that Ednrb signaling could act through the microRNA miR-196b to block the suppression of melanoblast proliferation by Aim2 in primary NC cell cultures. These results reveal the Ednrb–Aim2–AKT axis in regulating melanocyte development and suggest that Ednrb signaling functions as a negative regulator of Aim2, which inhibits the proliferation of melanoblasts in early development. These findings uncover a previously unreported role for Aim2 outside the inflammasome, showing that it is a significant regulator controlling NC stem cell-derived lineage development.

2024-11-06 | GSE275956 | GEO

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