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ctnnb1Dex3 mice die of heart failure between the second and fourth postnatal months. (A) Kaplan-Meier graph of survival of Tyr::CreA/u; ctnnb1Dex3/+ and WT littermate controls (Tyr::CreA/u; +/+ and u/u; ctnnb1Dex3/+). (B) Kaplan-Meier graph of survival of Tyr::CreB/u; ctnnb1Dex3/+ and WT littermate controls (Tyr::CreB/u; +/+). All members of both mutant populations perished before their fourth month of life, in contrast to the full survival of all their WT littermates. doi:10.1371/journal.pone.0053183.g006
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Background
Patent ductus arteriosus is a life-threatening condition frequent in premature newborns but also present in some term infants. Current mouse models of this malformation generally lead to perinatal death, not reproducing the full phenotypic spectrum in humans, in whom genetic inheritance appears complex. The ductus arteriosus (DA), a temp...
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Context 1
... mice have a greatly dilated left atrium, which develops after birth As expected and according to specific crosses, Tyr::Cre/u; ctnnb1Dex3/+ mice were produced in a Mendelian ratio (72 u/u; ctnnb1Dex3/+ versus 67 Tyr::Cre/u; ctnnb1Dex3/+ live births). However, all ctnnb1Dex3 mice died between 4 and 18 weeks of age ( Figure 6A). A distinct Tyr::Cre mouse line, Tyr::CreB, was used to see if the site of integration of the transgene or other cis- regulatory differences would affect the outcome [27]. ...
Context 2
... distinct Tyr::Cre mouse line, Tyr::CreB, was used to see if the site of integration of the transgene or other cis- regulatory differences would affect the outcome [27]. Nonetheless, all Tyr::CreB/u; ctnnb1Dex3/+ mice died within a similar age range (between 3 and 12 postnatal weeks; Figure 6B), indicating that the death of these animals was independent of the Tyr::Cre mouse line used. ...
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Multipotent stem cells with neural crest-like properties have been identified in the dermis of human skin. These neural crest stem cell (NCSC)-like cells display self-renewal capacity and differentiate into neural crest derivatives including epidermal pigment-producing melanocytes. NCSC-like cells share many properties with aggressive melanoma cell...
Citations
... The iris sphincter and dilator smooth muscles are derived from the neural crest of the optic cup. The ductus arteriosus is derived from the 6th aortic arch and its media contains neural crest smooth muscle cells in a spiral pattern with intervening endothelial cushions, enabling tight and permanent closure in response to oxygen in the postnatal period [13,14]. Disorders of these smooth muscle cells or their signalling are known to cause patent ductus, as seen in mutations of smooth muscle myosin (MYH11) and notch mutations of Alagille, Adams-Oliver and Hajdu-Cheney syndromes [15]. ...
The clinical and neuroimaging findings of a family with a variant ACTA2 gene (c351C > G), presenting with smooth muscle dysfunction in structures of neural crest derivation, are discussed. The combination of aortic abnormalities, patent ductus arteriosus, congenital mydriasis and distinctive cerebrovascular and brain morphological abnormalities characterise this disorder. Two sisters, heterozygous for the variant, and their mother, a mosaic, are presented. Brain parenchymal changes are detailed for the first time in a non-Arg179His variant. Radiological features of the petrous canal and external carotid are highlighted. We explore the potential underlying biological and embryological mechanisms.
... Among these were the adrenal or pituitary glands, the 180 latter of which control adrenal function through the function of its corticotropic cells. Subtle cardiac 181 malformations were an additional possibility: tricuspid valve insufficiency, leading to thickening of 182 ventricular or septal walls, or patent ductus arteriosus (Yajima et al., 2013). We therefore compared 183 these organ sizes on soft tissue contrast-enhanced micro-CT sections from E15.5-E20 fetuses (n=10 184 mutants vs 10 controls, Figure 2L). ...
Recurrent missense mutations of the PIK3CA oncogene are among the most frequent drivers of human cancers. These often lead to constitutive activation of its product p110α, a phosphatidylinositol 3-kinase (PI3K) catalytic subunit. In addition to causing a range of rare and common cancers, the H1047R mutation is also found in affected tissues of a distinct set of congenital tumours and malformations. Collectively termed PIK3CA -related disorders (PRDs), these lead to overgrowth of skin, brain, adipose, connective, musculoskeletal tissues and/or blood and lymphatic vessel components. Vascular malformations are frequently observed in PRD due to cell-autonomous activation of the PI3K signaling pathway within endothelial cells. These, like most muscle, connective tissue and bone, are derived from the embryonic mesoderm. However, important organ systems affected in PRDs are neuroectodermal derivatives. To further examine their development, we drove the most common post-zygotic activating mutation of Pik3ca in neural crest and related embryonic lineages. Effects in cells having once expressed Wnt1, including the brain roofplate and most neural crest, were most dramatic in the head. Outcomes included megalencephaly, cleft secondary palate and more subtle skull anomalies. Surprisingly, Pik3ca -mutant subpopulations of either mesodermal or neural crest origin was associated with widespread vascular anomalies, leading us to incidentally discover previously undescribed lineages that had expressed the transcription factor Egr2 (Krox20) and that may be co-opted in pathogenesis. Schwann cell precursors having transcribed either Krox20 or Sox10 also gave rise to adult-onset vascular tumors and cancers, including melanoma, after Pik3ca activation. These murine phenotypes may aid discovery of new candidate human PRDs affecting craniofacial and vascular smooth muscle development as well as the reciprocal paracrine signaling mechanisms leading to tissue overgrowth.
... In addition, increased proliferation of non-NCC-derived SMCs appeared as insufficient to compensate for reduced NCC-derived SMCs in Prdm6 fl/fl Wnt1-Cre2 ZsGreen1 mice. This finding is consistent with a prior report that NCC-derived and not mesodermal-derived SMCs are responsible for ductal closure (20). ...
... In response, we observed reduced migration of the delaminated NCCs compared with the PBS control, supporting our hypothesis (Figure 8, C and D). These findings are consistent with a prior report that constitutive activation of β-catenin in the NCC-derived SMC precursor results in a reduced NCC-derived population of SMCs in the DA (20), leading to its patency. Our findings also identified excess Wnt activation as a mechanism for PDA. ...
... Our investigations ex vivo demonstrated that increased Wnt1 underlies impaired CNCC migration in Prdm6-deficient mice, which was rescued by Wnt inhibition. These findings are consistent with a prior report that constitutive activation of β-catenin (ctnnb1Δex3) results in a reduced NCC-derived population of SMCs in the DA and early postnatal death (20). Although NCC migration in ctnnb1Δex3 was not explored, our fate mapping and functional studies suggest that excessive Wnt activation can result in impaired CNCC migration. ...
The molecular mechanisms that drive the acquisition of distinct neural crest cell (NCC) fates is still poorly understood. Here, we identify Prdm6 as an epigenetic modifier that temporally and spatially regulates the expression of NCC specifiers and determines the fate of a subset of migrating Cardiac NCCs (CNCCs). Using transcriptomic analysis, genetic and fate mapping approaches in transgenic mice, we show that disruption of Prdm6 is associated with impaired CNCC differentiation, delamination, and migration, and leads to patent ductus arteriosus (DA)and ventricular noncompaction. Bulk and single-cell RNA-seq analyses of DA and CNCC identify Prdm6 as a regulator of a network of CNCC specification genes including Wnt1, Tfap2b, and Sox9. Loss of Prdm6 in CNCCs diminishes its expression in pre-EMT cluster, resulting in the retention of NCC in the dorsal neural tube. This defect is associated with diminished H4K20 mono-methylation and G1-S progression and augmented Wnt1 transcript levels in pre-EMT and neural tube clusters, which we show is the major driver of the impaired CNCC migration. Altogether, these findings reveal Prdm6 as a key regulator of CNCC differentiation and migration and identify Prdm6 and its regulated network as potential targets for the treatment of congenital heart diseases.
... These results confirmed that DA SMCs derive from three populations: the nonpigmented non-VNCC-derived SMC1 (80%-90%), the non-pigmented VNCC-derived population SMC2 (10%-20%), and a very small number of pigmented VNCCderived melanoblasts (less than 1%). 26 Ctnnb1Δex3 mice exhibited shifts in cell population, with virtually all SMC2 cells replaced by melanoblasts and the SMC1 population unaffected. This shift was associated with PDA ( Figure 1F). ...
The ductus arteriosus (DA) is a unique fetal vascular shunt, which allows blood to bypass the developing lungs in utero. After birth, changes in complex signaling pathways lead to constriction and permanent closure of the DA. The persistent patency of the DA (PDA) is a common disorder in preterm infants, yet the underlying causes of PDA are not fully defined. Although limits on the availability of human DA tissues prevent comprehensive studies on the mechanisms of DA function, mouse models have been developed that reveal critical pathways in DA regulation. Over 20 different transgenic models of PDA in mice have been described, with implications for human DA biology. Similarly, we enumerate 224 human single‐gene syndromes that are associated with PDA, including a small subset that consistently feature PDA as a prominent phenotype. Comparison and functional analyses of these genes provide insight into DA development and identify key regulatory pathways that may serve as potential therapeutic targets for the management of PDA.
... (E) After removing the atria, one observes labeled great artery smooth muscle as well as at the base of the coronary arteries, the large branches of autonomic nerves and numerous melanocytes lining the right interatrial septum around the foramen ovale, all of NCC origin. Melanocytes are also frequently seen along valve leaflets, chordae tendinae, on and within the interventricular septum and within the proximal great arteries [132][133][134][135][136]. Anterior view, ventral to top. ...
Anomalies in the cardiac outflow tract (OFT) are among the most frequent congenital heart defects (CHDs). During embryogenesis, the cardiac OFT is a dynamic structure at the arterial pole of the heart. Heart tube elongation occurs by addition of cells from pharyngeal, splanchnic mesoderm to both ends. These progenitor cells, termed the second heart field (SHF), were first identified twenty years ago as essential to the growth of the forming heart tube and major contributors to the OFT. Perturbation of SHF development results in common forms of CHDs, including anomalies of the great arteries. OFT development also depends on paracrine interactions between multiple cell types, including myocardial, endocardial and neural crest lineages. In this publication, dedicated to Professor Andriana Gittenberger-De Groot and her contributions to the field of cardiac development and CHDs, we review some of her pioneering studies of OFT development with particular interest in the diverse origins of the many cell types that contribute to the OFT. We also discuss the clinical implications of selected key findings for our understanding of the etiology of CHDs and particularly OFT malformations.
... To confirm that the observed reduction of perinuclear melanosomes in this mutant cell line was not limited to the specific bcat* construct, we expressed another active β-catenin mutant, the βcat-Δex3-GFP mutant, in WT melanocytes 37 . This mutant construct encodes for a β-catenin protein lacking the serine and threonine residues that allow for its proteasomal degradation. ...
... Transfections were assayed 48 h following nucleofection. Control GFP and β-catenin-GFP-tagged plasmids have been previously described 37 . The Dynlt3-GFP (pCMV3-mDynlt3-GFPSpark) and the control GFP vectors (pCMV3-GFP) were purchased from Sino Biological (MG5A2492-ACG and CV026, respectively). ...
Skin pigmentation is dependent on cellular processes including melanosome biogenesis, transport, maturation and transfer to keratinocytes. However, how the cells finely control these processes in space and time to ensure proper pigmentation remains unclear. Here, we show that a component of the cytoplasmic dynein complex, Dynlt3, is required for efficient melanosome transport, acidity and transfer. In Mus musculus melanocytes with decreased levels of Dynlt3, pigmented melanosomes undergo a more directional motion, leading to their peripheral location in the cell. Stage IV melanosomes are more acidic, but still heavily pigmented, resulting in a less efficient melanosome transfer. Finally, the level of Dynlt3 is dependent on β-catenin activity, revealing a function of the Wnt/β-catenin signalling pathway during melanocyte and skin pigmentation, by coupling the transport, positioning and acidity of melanosomes required for their transfer.
... To confirm that the observed reduction of perinuclear melanosomes in this mutant cell line was not limited to the specific bcat* construct, we expressed another active β-catenin mutant, the βcat-Δex3-GFP mutant, in WT melanocytes 37 . This mutant construct encodes for a β-catenin protein lacking the serine and threonine residues that allow for its proteasomal degradation. ...
... Transfections were assayed 48 h following nucleofection. Control GFP and β-catenin-GFP-tagged plasmids have been previously described 37 . The Dynlt3-GFP (pCMV3-mDynlt3-GFPSpark) and the control GFP vectors (pCMV3-GFP) were purchased from Sino Biological (MG5A2492-ACG and CV026, respectively). ...
Skin pigmentation is dependent on cellular processes including melanosome biogenesis, transport, maturation and transfer to keratinocytes. However, how the cells finely control these processes in space and time to ensure proper pigmentation remains unclear. Here, we show that a component of the cytoplasmic dynein complex, Dynlt3, is required for efficient melanosome transport, maturation and transfer. In melanocytes with decreased levels of Dynlt3, pigmented melanosomes undergo a more directional (convective) motion leading to their peripheral location in the cell, and are not fully matured. Stage IV melanosomes are more acidic, but still heavily pigmented, resulting in a less efficient melanosome transfer. Finally, the level of Dynlt3 is dependent on β-catenin activity, revealing a novel function of the Wnt/β-catenin signalling pathway during melanocyte and skin pigmentation, by coupling the transport, position and maturation of melanosomes required for their transfer.
... We only compared low birth weight group without including normal weight group. Low birth weight plays role in the pathophysiology of PDA through several mechanisms such as: impaired differentiation, proliferation, migration and contractility of ductus arteriosus due to lack of smooth muscle cell, resulting in failure of ductus arteriosus closure while the baby was borned [7,11]; the differences in the anatomical structure of ductus arteriosus walls and lumen in normal birth weight infant and infant with low birth weight [12]. ...
... To explore in vivo Lkb1 functions during lineage establishment from NCCs, Lkb1 conditional mutant mice (Lkb1 F/F ) (19) were interbred to the Tyr::Cre deleter ( [20][21][22] and the R26R reporter (23) (20,21,24). A role of Lkb1 in Schwann cell myelination and axon maintenance has been previously reported (25)(26)(27). ...
Metabolic processes underlying the development of the neural crest, an embryonic population of multipotent migratory cells, are poorly understood. Here, we report that conditional ablation of the Lkb1 tumor suppressor kinase in mouse neural crest stem cells led to intestinal pseudo-obstruction and hind limb paralysis. This phenotype originated from a postnatal degeneration of the enteric nervous ganglia and from a defective differentiation of Schwann cells. Metabolomic profiling revealed that pyruvate-alanine conversion is enhanced in the absence of Lkb1 . Mechanistically, inhibition of alanine transaminases restored glial differentiation in an mTOR-dependent manner, while increased alanine level directly inhibited the glial commitment of neural crest cells. Treatment with the metabolic modulator AICAR suppressed mTOR signaling and prevented Schwann cell and enteric defects of Lkb1 mutant mice. These data uncover a link between pyruvate-alanine cycling and the specification of glial cell fate with potential implications in the understanding of the molecular pathogenesis of neural crest diseases.
... However, our DNA microarray data indicated that gene expression profiles between the proximal and distal ductus arteriosi were much closer than that of the aorta. It is known that the smooth muscle cells of DA are derived entirely from cardiac neural crest cells, whereas smooth muscle cells of the aorta are primarily derived from mesenchymal cells [21][22][23]. This may account for the similarity of transcriptional profiles between the proximal and distal ductus arteriosi when compared to that of the descending aorta. ...
The ductus arteriosus, an essential embryonic blood vessel between the pulmonary artery and the descending aorta, constricts after birth or hatching and eventually closes to terminate embryonic circulation. Chicken embryos have two long ductus arteriosi, which anatomically differ from mammal ductus arteriosus. Each long ductus arteriosus is divided into two parts: the pulmonary artery-sided and descending aorta-sided ductus arteriosi. Although the pulmonary artery-sided and descending aorta-sided ductus arteriosi have distinct functional characteristics, such as oxygen responsiveness, the difference in their transcriptional profiles has not been investigated. We performed a DNA microarray analysis (GSE 120116 at NCBI GEO) with pooled tissues from the chicken pulmonary artery-sided ductus arteriosus, descending aorta-sided ductus arteriosus, and aorta at the internal pipping stage. Although several known ductus arteriosus-dominant genes such as tfap2b were highly expressed in the pulmonary artery-sided ductus arteriosus, we newly found genes that were dominantly expressed in the chicken pulmonary artery-sided ductus arteriosus. Interestingly, cluster analysis showed that the expression pattern of the pulmonary artery-sided ductus arteriosus was closer to that of the descending aorta-sided ductus arteriosus than that of the aorta, whereas the morphology of the descending aorta-sided ductus arteriosus was closer to that of the aorta than that of the pulmonary artery-sided ductus arteriosus. Subsequent pathway analysis with DAVID bioinformatics resources revealed that the pulmonary artery-sided ductus arteriosus showed enhanced expression of the genes involved in melanogenesis and tyrosine metabolism compared with the descending aorta-sided ductus arteriosus, suggesting that tyrosinase and the related genes play an important role in the proper differentiation of neural crest-derived cells during vascular remodeling in the ductus arteriosus. In conclusion, the transcription profiles of the chicken ductus arteriosus provide new insights for investigating the mechanism of ductus arteriosus closure.
