Mutations in cdon and boc affect trunk neural crest cell migration and slow-twitch muscle development in zebrafish
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ABSTRACT:
ORGANISM(S): Danio rerio
PROVIDER: GSE213728 | GEO | 2023/07/03
REPOSITORIES: GEO
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Project description:Mutations in cdon and boc affect trunk neural crest cell migration and slow-twitch muscle development in zebrafish
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Project description:Neural crest cells migrate extensively in vertebrate embryos to populate diverse derivatives including ganglia of the peripheral nervous system. Little is known about the molecular mechanisms that tell migrating trunk neural crest cells to settle at selected sites in the embryo by ceasing migration and initiating differentiation programs.
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Project description:Ectoderm-derived neural crest is a transient structure arising during early embryogenesis in vertebrates. Neural crest consists of four derivatives based on their anterior- to posterior location along the body axis; cranial, vagal, trunk and sacral, respectively. We recently showed that trunk neural crest-specific gene MOXD1 functions as a tumor suppressor in trunk neural crest-derived childhood cancer form neuroblastoma and is essential for proper development of healthy adrenal glands. However, the role of MOXD1 during early embryogenesis is not known. Here, we conditionally knocked out MOXD1 in trunk neural crest cells before they become lineage-committed, using a CRISPR/Cas9 approach in chick embryos. Assessment of embryo growth showed that knockout of MOXD1 delayed development with knockout embryos being smaller. RNA sequencing of trunk-derived neural crest cells from control and knockout embryos showed enrichment of genes connected to gland development, copper ion metabolism and neuroblastoma progression. In conclusion, MOXD1 is important during early and prolonged embryonic development with effects on gland formation, possibly mediated via its role in copper metabolism.
2023-09-13 | GSE242700 | GEO
Project description:The neural crest (NC) is a major contributor to the vertebrate craniofacial skeleton, detailed in model organisms through embryological and genetic approaches, most notably in chick and mouse. Despite many similarities between these rather distant species, there are also distinct differences in the contribution of the NC, particularly to the calvariae of the skull. Lack of information about other vertebrate groups precludes an understanding of the evolutionary significance of these differences. Study of zebrafish craniofacial development has contributed substantially to understanding of cartilage and bone formation in teleosts, but there is currently little information on NC contribution to the zebrafish skeleton. Here, we employ a two-transgene system based on Cre recombinase to genetically label NC in the zebrafish. We demonstrate NC contribution to cells in the cranial ganglia and peripheral nervous system known to be NC-derived, as well as to a subset of myocardial cells. The indelible labeling also enables us to determine NC contribution to late-forming bones, including the calvariae. We confirm suspected NC origin of cartilage and bones of the viscerocranium, including cartilages such as the hyosymplectic and its replacement bones (hymandibula and symplectic) and membranous bones such as the opercle. The cleithrum develops at the border of NC and mesoderm, and as an ancestral component of the pectoral girdle was predicted to be a hybrid bone composed of both NC and mesoderm tissues. However, we find no evidence of a NC contribution to the cleithrum. Similarly, in the vault of the skull, the parietal bones and the caudal portion of the frontal bones show no evidence of NC contribution. We also determine a NC origin for caudal fin lepidotrichia; the presumption is that these are derived from trunk NC, demonstrating that these cells have the ability to form bone during normal vertebrate development.
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Project description:Bone is an evolutionary novelty of vertebrates, likely to have first emerged as part of ancestral dermal armor that consisted of osteogenic and odontogenic components. Whether these early vertebrate structures arose from mesoderm or neural crest cells has been a matter of considerable debate. To examine the developmental origin of the bony part of the dermal armor, we have performed in vivo lineage tracing in the sterlet sturgeon, a representative of non-teleost ray-finned fish that has retained an extensive postcranial dermal skeleton. The results definitively show that sterlet trunk neural crest cells give rise to osteoblasts of the scutes. Transcriptional profiling further reveals neural crest gene signature in sterlet scutes as well as bichir scales. Finally, histological and microCT analysis of ray-finned fish dermal armor show that their scales and scutes are formed by bone, dentin and hypermineralized covering tissues, in various combinations, that resemble those of the first armored vertebrates. Taken together, our results support a primitive skeletogenic role for the neural crest along the entire body axis, that was later progressively restricted to the cranial region during vertebrate evolution. Thus, the neural crest was a crucial evolutionary innovation driving the origin and diversification of dermal armor along the entire body axis.
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Project description:We report that cancer associated protein HIF-2a is expressed in trunk neural crest neuroblastoma precursor cells in the developing embryo in three different species; human, mouse and avian. Dysregulation of HIF-2a leads to alterations in embryonic development, and neural crest cell migration, proliferation and self-renewal capacity. With RNAsequencing we report that alterations of HIF-2a expression affects the global transcriptome and that gene ontology enrich for the same processes observed in vivo.
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Project description:CHARGE syndrome is caused by heterozygous mutations in a chromatin remodeler CHD7 and characterized by a set of malformations historically postulated to arise from defects in the neural crest formation during embryogenesis. To better delineate neural crest defects in CHARGE syndrome, we generated induced pluripotent stem cells (iPSCs) from two patients with typical syndrome manifestations, and characterized neural crest cells differentiated in vitro from these iPSCs (iPSC-NCCs). We found that expression of genes associated with cell migration was altered in CHARGE iPSC-NCCs as compared to control iPSC-NCCs. Consistently, CHARGE iPSC-NCCs showed defective delamination, migration and motility in vitro, and their transplantation in ovo revealed overall defective migratory activity in the chick embryo. Altogether, our results support the historical inference that CHARGE syndrome patients have defects in neural crest migration and provide the first successful application of patient-derived iPSCs in modeling craniofacial disorders.
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Project description:Neural crest (NC) cells undergo extensive migrations during development. Here, we couple in vivo live imaging at high resolution with custom software tools to reveal dynamic migratory behavior in chick embryos. Trunk NC cells migrate as individuals with both stochastic and biased features as they move dorsoventrally to form peripheral ganglia. Their leading edge displays a prominent fan-shaped lamellipodium that reorients upon cell-cell contact. Computational analysis reveals that when the lamellipodium of one cell touches the body of another, the two cells undergo "contact attraction," often moving together and then separating via a pulling force exerted by lamellipodium. Targeted optical manipulation shows that cell interactions coupled with cell density generate a long-range biased random walk behavior, such that cells move from high to low density. In contrast to chain migration noted at other axial levels, the results show that individual trunk NC cells navigate the complex environment without tight coordination between neighbors.
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Project description:Single cell RNA analysis of trunk neural crest cells in zebrafish
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