RESUMO
Apical expansion of calvarial osteoblast progenitors from the cranial mesenchyme (CM) above the eye is integral to calvarial growth and enclosure of the brain. The cellular behaviors and signals underlying the morphogenetic process of calvarial expansion are unknown. Time-lapse light-sheet imaging of mouse embryos revealed calvarial progenitors intercalate in 3D in the CM above the eye, and exhibit protrusive and crawling activity more apically. CM cells express non-canonical Wnt/planar cell polarity (PCP) core components and calvarial osteoblasts are bidirectionally polarized. We found non-canonical ligand Wnt5a-/- mutants have less dynamic cell rearrangements and protrusive activity. Loss of CM-restricted Wntless (CM-Wls), a gene required for secretion of all Wnt ligands, led to diminished apical expansion of Osx+ calvarial osteoblasts in the frontal bone primordia in a non-cell autonomous manner without perturbing proliferation or survival. Calvarial osteoblast polarization, progressive cell elongation and enrichment for actin along the baso-apical axis were dependent on CM-Wnts. Thus, CM-Wnts regulate cellular behaviors during calvarial morphogenesis for efficient apical expansion of calvarial osteoblasts. These findings also offer potential insights into the etiologies of calvarial dysplasias.
Assuntos
Mesoderma , Morfogênese , Osteoblastos , Crânio , Proteínas Wnt , Animais , Osteoblastos/metabolismo , Osteoblastos/citologia , Crânio/embriologia , Camundongos , Mesoderma/citologia , Mesoderma/metabolismo , Proteínas Wnt/metabolismo , Proteínas Wnt/genética , Polaridade Celular , Proteína Wnt-5a/metabolismo , Proteína Wnt-5a/genética , Movimento Celular , Proliferação de CélulasRESUMO
The skull roof, or calvaria, is comprised of interlocking plates of bones that encase the brain. Separating these bones are fibrous sutures that permit growth. Currently, we do not understand the instructions for directional growth of the calvaria, a process which is error-prone and can lead to skeletal deficiencies or premature suture fusion (craniosynostosis, CS). Here, we identify graded expression of fibronectin (FN1) in the mouse embryonic cranial mesenchyme (CM) that precedes the apical expansion of calvaria. Conditional deletion of Fn1 or Wasl leads to diminished frontal bone expansion by altering cell shape and focal actin enrichment, respectively, suggesting defective migration of calvarial progenitors. Interestingly, Fn1 mutants have premature fusion of coronal sutures. Consistently, syndromic forms of CS in humans exhibit dysregulated FN1 expression, and we also find FN1 expression altered in a mouse CS model of Apert syndrome. These data support a model of FN1 as a directional substrate for calvarial osteoblast migration that may be a common mechanism underlying many cranial disorders of disparate genetic etiologies.
Assuntos
Fibronectinas , Nascimento Prematuro , Crânio , Animais , Feminino , Humanos , Camundongos , Sinais (Psicologia) , Modelos Animais de Doenças , Fibronectinas/metabolismo , Osteoblastos , Crânio/citologia , Crânio/crescimento & desenvolvimento , Crânio/metabolismo , SuturasRESUMO
Skin is largely composed of an epidermis that overlies a supporting dermis. Recent advancements in our understanding of how diverse groups of dermal fibroblasts regulate epidermal and hair follicle growth and differentiation have been fueled by tools capable of resolving molecular heterogeneity at a single-cell level. Fibroblast heterogeneity can be traced back to their developmental origin before their segregation into spatially distinct fibroblast subtypes. The mechanisms that drive this lineage diversification during development are being unraveled, with studies showing that both large- and small-scale positional signals play important roles during dermal development. Here, we first delineate what is known about the origins of the dermis and the central role of Wnt/ß-catenin signaling in its specification across anatomical locations. We then discuss how one of the first morphologically recognizable fibroblast subtypes, the hair follicle dermal condensate lineage, emerges. Leveraging the natural variation of skin and its appendages between species and between different anatomical locations, these collective studies have identified shared and divergent factors that contribute to the extraordinary diversity of skin.
Assuntos
Epiderme , Pele , Folículo Piloso , Fibroblastos , Células EpidérmicasRESUMO
Fibrosis is primarily described as the deposition of excessive extracellular matrix, but in many tissues it also involves a loss of lipid or lipid-filled cells. Lipid-filled cells are critical to tissue function and integrity in many tissues including the skin and lungs. Thus, loss or depletion of lipid-filled cells during fibrogenesis, has implications for tissue function. In some contexts, lipid-filled cells can impact ECM composition and stability, highlighting their importance in fibrotic transformation. Recent papers in fibrosis address this newly recognized fibrotic lipodystrophy phenomenon. Even in disparate tissues, common mechanisms are emerging to explain fibrotic lipodystrophy. These findings have implications for fibrosis in tissues composed of fibroblast and lipid-filled cell populations such as skin, lung, and liver. In this review, we will discuss the roles of lipid-containing cells, their reduction/loss during fibrotic transformation, and the mechanisms of that loss in the skin and lungs.
Assuntos
Lipodistrofia , Pele , Humanos , Fibrose , Pele/patologia , Pulmão/patologia , Matriz Extracelular/patologia , Fibroblastos/patologia , Lipodistrofia/patologia , LipídeosRESUMO
Skin development and patterning is dependent on factors that regulate the stepwise differentiation of dermal fibroblasts concomitant with dermal-epidermal reciprocal signaling, two processes that are poorly understood. Here we show that dermal EZH2, the methyltransferase enzyme of the epigenetic Polycomb Repressive Complex 2 (PRC2), is a new coordinator of both these processes. Dermal EZH2 activity is present during dermal fibroblast differentiation and is required for spatially restricting Wnt/ß-catenin signaling to reinforce dermal fibroblast cell fate. Later in development, dermal EZH2 regulates the expression of reticular dermal markers and initiation of secondary hair follicles. Embryos lacking dermal Ezh2 have elevated epidermal proliferation and differentiation that can be rescued by small molecule inhibition of retinoic acid (RA) signaling. Together, our study reveals that dermal EZH2 is acting like a rheostat to control the levels of Wnt/ß-catenin and RA signaling to impact fibroblast differentiation cell autonomously and epidermal keratinocyte development non-cell autonomously, respectively.
Assuntos
Derme/citologia , Derme/embriologia , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Epiderme/embriologia , Fibroblastos/citologia , Queratinócitos/citologia , Complexo Repressor Polycomb 2/metabolismo , Animais , Diferenciação Celular , Proliferação de Células , Derme/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Epiderme/metabolismo , Fibroblastos/metabolismo , Hiperplasia , Queratinócitos/metabolismo , Camundongos , Organogênese , Retinoides/farmacologia , Transdução de Sinais , Células-Tronco/citologia , Células-Tronco/metabolismo , Tretinoína/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismoRESUMO
PURPOSE OF REVIEW: Epigenetic regulation is a distinct mechanism of gene regulation that functions by modulating chromatin structure and accessibility. Polycomb Repressive Complex 2 (PRC2) is a conserved chromatin regulator that is required in the developing embryo to control the expression of key developmental genes. An emerging feature of PRC2 is that it not only allows for binary ON/OFF states of gene expression but can also modulate gene expression in feed-forward loops to change the outcome of gene regulatory networks. This striking feature of epigenetic modulation has improved our understanding of musculoskeletal development. RECENT FINDINGS: Recent advances in mouse embryos unravel a range of phenotypes that demonstrate the tissue-specific, temporal, and spatial role of PRC2 during organogenesis and cell fate decisions in vivo. Here, we take a detailed view of how PRC2 functions during the development of calvarial bone and skin. Based on the emerging evidence, we propose that PRC2 serves as a "dimmer switch" to modulate gene expression of target genes by altering the expression of activators and inhibitors. This review highlights the findings from contemporary research that allow us to investigate the unique developmental potential of intramembranous calvarial bones.
Assuntos
Desenvolvimento Ósseo/genética , Epiderme/embriologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Complexo Repressor Polycomb 2/genética , Crânio/embriologia , Animais , Humanos , CamundongosRESUMO
The formation of the head and face is a complex process which involves many different signaling cues regulating the migration, differentiation, and proliferation of the neural crest. This highly complex process is very error-prone, resulting in craniofacial defects in nearly 10,000 births in the United States annually. Due to the highly conserved mechanisms of craniofacial development, animal models are widely used to understand the pathogenesis of various human diseases and assist in the diagnosis and generation of preventative therapies and treatments. Here, we provide a brief background of craniofacial development and discuss several rare diseases affecting craniofacial bone development. We focus on rare congenital diseases of the cranial bone, facial jaw bones, and two classes of diseases, ciliopathies and RASopathies. Studying the animal models of these rare diseases sheds light not only on the etiology and pathology of each disease, but also provides meaningful insights towards the mechanisms which regulate normal development of the head and face.
Assuntos
Anormalidades Craniofaciais , Modelos Animais de Doenças , Cabeça/embriologia , Animais , Anormalidades Craniofaciais/diagnóstico , Anormalidades Craniofaciais/prevenção & controle , Anormalidades Craniofaciais/terapia , Face/embriologia , Humanos , Crista Neural/embriologia , Crânio/embriologiaRESUMO
Cranial neural crest cells (CNCCs) give rise to cranial mesenchyme (CM) that differentiates into the forebrain meningeal progenitors in the basolateral and apical regions of the head. This occurs in close proximity to the other CNCC-CM-derivatives, such as calvarial bone and dermal progenitors. We found active Wnt signaling transduction in the forebrain meningeal progenitors in basolateral and apical populations and in the non-meningeal CM preceding meningeal differentiation. Here, we dissect the source of Wnt ligand secretion and requirement of Wnt/ß-catenin signaling for the lineage selection and early differentiation of the forebrain meninges. We find persistent canonical Wnt/ß-catenin signal transduction in the meningeal progenitors in the absence of Wnt ligand secretion in the CM or surface ectoderm, suggesting additional sources of Wnts. Conditional mutants for Wntless and ß-catenin in the CM showed that Wnt ligand secretion and Wnt/ß-catenin signaling were dispensable for specification and proliferation of early meningeal progenitors. In the absence of ß-catenin in the CM, we found diminished laminin matrix and meningeal hypoplasia, indicating a structural and trophic role of mesenchymal ß-catenin signaling. This study shows that ß-catenin signaling is required in the CM for maintenance and organization of the differentiated meningeal layers in the basolateral and apical populations of embryonic meninges.
Assuntos
Meninges/embriologia , Mesoderma/metabolismo , Via de Sinalização Wnt , Animais , Linhagem da Célula , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Meninges/citologia , Meninges/metabolismo , Mesoderma/citologia , Mesoderma/embriologia , Camundongos , Prosencéfalo/citologia , Prosencéfalo/embriologia , Prosencéfalo/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , beta Catenina/genética , beta Catenina/metabolismoRESUMO
The skull bones must grow in a coordinated, three-dimensional manner to coalesce and form the head and face. Mammalian skull bones have a dual embryonic origin from cranial neural crest cells (CNCC) and paraxial mesoderm (PM) and ossify through intramembranous ossification. The calvarial bones, the bones of the cranium which cover the brain, are derived from the supraorbital arch (SOA) region mesenchyme. The SOA is the site of frontal and parietal bone morphogenesis and primary center of ossification. The objective of this review is to frame our current in vivo understanding of the morphogenesis of the calvarial bones and the gene networks regulating calvarial bone initiation in the SOA mesenchyme.
Assuntos
Desenvolvimento Ósseo , Regulação da Expressão Gênica no Desenvolvimento , Crânio/embriologia , Animais , Epigênese Genética , Humanos , Crânio/metabolismoRESUMO
Development of the skull bones requires the coordination of two stem progenitor populations, the cranial neural crest cells (CNCC) and head paraxial mesoderm (PM), to ensure cell fate selection and morphogenesis. The epigenetic methyltransferase, Ezh2, plays a role in skull bone formation, but the spatiotemporal function of Ezh2 between the CNCC- and PM-derived bone formation in vivo remains undefined. Here, using a temporally-inducible conditional deletion of Ezh2 in both the CNCC- and PM- derived cranial mesenchyme between E8.5 and E9.5, we find a reduction of the CNCC-derived calvarial bones and a near complete loss of the PM-derived calvarial bones due to an arrest in calvarial bone fate commitment. In contrast, deletion of Ezh2 after E9.5 permits PM-derived skull bone development, suggesting that Ezh2 is required early to guide calvarial bone progenitor commitment. Furthermore, exposure to all-trans Retinoic acid at E10.0 can mimic the Ezh2 mutant calvarial phenotype, and administration of the pan retinoic acid receptor (RAR) antagonist, BMS-453, to Ezh2 mutants partially restores the commitment to the calvarial bone lineage and PM-derived bone development in vivo. Exogenous RA signaling activation in the Ezh2 mutants leads to synergistic activation of the anti-osteogenic factors in the cranial mesenchyme in vivo. Thus, RA signaling and EZH2 can function in parallel to guide calvarial bone progenitor commitment by balancing the suppression of anti-osteogenic factors.
Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Crânio/embriologia , Tretinoína/metabolismo , Animais , Diferenciação Celular/genética , Linhagem da Célula/genética , Proteína Potenciadora do Homólogo 2 de Zeste/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Idade Gestacional , Mesoderma/embriologia , Mesoderma/metabolismo , Camundongos , Crista Neural/embriologia , Crista Neural/metabolismo , Transdução de Sinais , Crânio/metabolismo , Tretinoína/fisiologiaRESUMO
The Sonic hedgehog (Shh) signaling pathway regulates developmental, homeostatic, and repair processes throughout the body. In the skin, touch domes develop in tandem with primary hair follicles and contain sensory Merkel cells. The developmental signaling requirements for touch dome specification are largely unknown. We found dermal Wnt signaling and subsequent epidermal Eda/Edar signaling promoted Merkel cell morphogenesis by inducing Shh expression in early follicles. Lineage-specific gene deletions revealed intraepithelial Shh signaling was necessary for Merkel cell specification. Additionally, a Shh signaling agonist was sufficient to rescue Merkel cell differentiation in Edar-deficient skin. Moreover, Merkel cells formed in Fgf20 mutant skin where primary hair formation was defective but Shh production was preserved. Although developmentally associated with hair follicles, fate mapping demonstrated Merkel cells primarily originated outside the hair follicle lineage. These findings suggest that touch dome development requires Wnt-dependent mesenchymal signals to establish reciprocal signaling within the developing ectoderm, including Eda signaling to primary hair placodes and ultimately Shh signaling from primary follicles to extrafollicular Merkel cell progenitors. Shh signaling often demonstrates pleiotropic effects within a structure over time. In postnatal skin, Shh is known to regulate the self-renewal, but not the differentiation, of touch dome stem cells. Our findings relate the varied effects of Shh in the touch dome to the ligand source, with locally produced Shh acting as a morphogen essential for lineage specification during development and neural Shh regulating postnatal touch dome stem cell maintenance.
Assuntos
Ectodisplasinas/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog/metabolismo , Células de Merkel/citologia , Proteína Wnt1/metabolismo , Animais , Linhagem da Célula , Reparo do DNA , Feminino , Fatores de Crescimento de Fibroblastos/metabolismo , Deleção de Genes , Genótipo , Folículo Piloso/embriologia , Folículo Piloso/metabolismo , Homeostase , Ligantes , Masculino , Camundongos , Microscopia de Fluorescência , Morfogênese , Mutação , Neurônios/metabolismo , Transdução de Sinais , Pele/embriologia , Pele/metabolismo , TatoRESUMO
The cranial bones and dermis differentiate from mesenchyme beneath the surface ectoderm. Fate selection in cranial mesenchyme requires the canonical Wnt effector molecule ß-catenin, but the relative contribution of Wnt ligand sources in this process remains unknown. Here we show Wnt ligands are expressed in cranial surface ectoderm and underlying supraorbital mesenchyme during dermal and osteoblast fate selection. Using conditional genetics, we eliminate secretion of all Wnt ligands from cranial surface ectoderm or undifferentiated mesenchyme, to uncover distinct roles for ectoderm- and mesenchyme-derived Wnts. Ectoderm Wnt ligands induce osteoblast and dermal fibroblast progenitor specification while initiating expression of a subset of mesenchymal Wnts. Mesenchyme Wnt ligands are subsequently essential during differentiation of dermal and osteoblast progenitors. Finally, ectoderm-derived Wnt ligands provide an inductive cue to the cranial mesenchyme for the fate selection of dermal fibroblast and osteoblast lineages. Thus two sources of Wnt ligands perform distinct functions during osteoblast and dermal fibroblast formation.
Assuntos
Diferenciação Celular/genética , Crânio/crescimento & desenvolvimento , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Animais , Ectoderma/crescimento & desenvolvimento , Ectoderma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Ligantes , Mesoderma/citologia , Mesoderma/crescimento & desenvolvimento , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Transdução de Sinais , Crânio/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
BACKGROUND: Specification of cranial bone and dermal fibroblast progenitors in the supraorbital arch mesenchyme is Wnt/ß-catenin signaling-dependent. The mechanism underlying how these cells interpret instructive signaling cues and differentiate into these two lineages is unclear. Twist1 is a target of the Wnt/ß-catenin signaling pathway and is expressed in cranial bone and dermal lineages. RESULTS: Here, we show that onset of Twist1 expression in the mouse cranial mesenchyme is dependent on ectodermal Wnts and mesenchymal ß-catenin activity. Conditional deletion of Twist1 in the supraorbital arch mesenchyme leads to cranial bone agenesis and hypoplastic dermis, as well as craniofacial malformation of eyes and palate. Twist1 is preferentially required for cranial bone lineage commitment by maintaining Wnt responsiveness. In the conditional absence of Twist1, the cranial dermis fails to condense and expand apically leading to extensive cranial dermal hypoplasia with few and undifferentiated hair follicles. CONCLUSIONS: Thus, Twist1, a target of canonical Wnt/ß-catenin signaling, also functions to maintain Wnt responsiveness and is a key effector for cranial bone fate selection and dermal condensation.
Assuntos
Anormalidades Craniofaciais/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Nucleares/metabolismo , Crânio/embriologia , Proteína 1 Relacionada a Twist/metabolismo , Via de Sinalização Wnt/fisiologia , Animais , Anormalidades Craniofaciais/genética , Folículo Piloso/metabolismo , Camundongos , Camundongos Knockout , Proteínas Nucleares/genética , Crânio/metabolismo , Proteína 1 Relacionada a Twist/genética , Proteínas Wnt/metabolismo , beta Catenina/metabolismoRESUMO
Embryonic dermal fibroblasts in the skin have the exceptional ability to initiate hair follicle morphogenesis and contribute to scarless wound healing. Activation of the Wnt signaling pathway is critical for dermal fibroblast fate selection and hair follicle induction. In humans, mutations in Wnt pathway components and target genes lead to congenital focal dermal hypoplasias with diminished hair. The gene expression signature of embryonic dermal fibroblasts during differentiation and its dependence on Wnt signaling is unknown. Here we applied Shannon entropy analysis to identify the gene expression signature of mouse embryonic dermal fibroblasts. We used available human DNase-seq and histone modification ChiP-seq data on various cell-types to demonstrate that genes in the fibroblast cell identity signature can be epigenetically repressed in other cell-types. We found a subset of the signature genes whose expression is dependent on Wnt/ß-catenin activity in vivo. With our approach, we have defined and validated a statistically derived gene expression signature that may mediate dermal fibroblast identity and function in development and disease. genesis 54:415-430, 2016. © 2016 Wiley Periodicals, Inc.
Assuntos
Diferenciação Celular , Derme/citologia , Fibroblastos/metabolismo , Transcriptoma , Via de Sinalização Wnt/genética , Animais , Derme/embriologia , Fibroblastos/citologia , Regulação da Expressão Gênica no Desenvolvimento , CamundongosRESUMO
The ERK/MAPK pathway is an important developmental signaling pathway. Mutations in upstream elements of this pathway result in neuro-cardio-facial cutaneous (NCFC) syndromes, which are typified by impaired neurocognitive abilities that are reliant upon hippocampal function. The role of ERK signaling during hippocampal development has not been examined and may provide critical insight into the cause of hippocampal dysfunction in NCFC syndromes. In this study, we have generated ERK1 and conditional ERK2 compound knock-out mice to determine the role of ERK signaling during development of the hippocampal dentate gyrus. We found that loss of both ERK1 and ERK2 resulted in 60% fewer granule cells and near complete absence of neural progenitor pools in the postnatal dentate gyrus. Loss of ERK1/2 impaired maintenance of neural progenitors as they migrate from the dentate ventricular zone to the dentate gyrus proper, resulting in premature depletion of neural progenitor cells beginning at E16.5, which prevented generation of granule cells later in development. Finally, loss of ERK2 alone does not impair development of the dentate gyrus as animals expressing only ERK1 developed a normal hippocampus. These findings establish that ERK signaling regulates maintenance of progenitor cells required for development of the dentate gyrus.
Assuntos
Giro Denteado , Retroalimentação Fisiológica/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Células-Tronco/fisiologia , Animais , Animais Recém-Nascidos , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Giro Denteado/embriologia , Giro Denteado/enzimologia , Giro Denteado/crescimento & desenvolvimento , Embrião de Mamíferos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Ventrículos Laterais/citologia , Ventrículos Laterais/embriologia , Ventrículos Laterais/crescimento & desenvolvimento , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neurogênese/genética , Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-raf/genética , Proteínas Proto-Oncogênicas c-raf/metabolismoRESUMO
Fibrosis is an end-stage response to tissue injury that is associated with loss of organ function as a result of excess extracellular matrix (ECM) production by fibroblasts. In skin, pathological fibrosis is evident during keloid scar formation, systemic sclerosis (SSc) and morphea. Dermal fibroblasts in these fibrotic diseases exhibit increased Wnt/ß-catenin signalling, a pathway that is sufficient to cause fibrosis in mice. However, in the context of this complex pathology, the precise pro-fibrotic consequences of Wnt/ß-catenin signalling are not known. We found that expression of stabilized ß-catenin in mouse dermal fibroblasts resulted in spontaneous, progressive skin fibrosis with thickened collagen fibres and altered collagen fibril morphology. The fibrotic phenotype was predominated by resident dermal fibroblasts. Genome-wide profiling of the fibrotic mouse dermis revealed elevated expression of matrix-encoding genes, and the promoter regions of these genes were enriched for Tcf/Lef family transcription factor binding sites. Additionally, we identified 32 ß-catenin-responsive genes in our mouse model that are also over-expressed in human fibrotic tissues and poised for regulation by Tcf/Lef family transcription factors. Therefore, we have uncovered a matrix-regulatory role for stabilized ß-catenin in fibroblasts in vivo and have defined a set of ß-catenin-responsive genes with relevance to fibrotic disease.
Assuntos
Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Pele/metabolismo , beta Catenina/metabolismo , Animais , Sítios de Ligação , Proteínas da Matriz Extracelular/genética , Fibroblastos/patologia , Fibrose , Perfilação da Expressão Gênica/métodos , Genótipo , Humanos , Camundongos Transgênicos , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Regiões Promotoras Genéticas , Pele/patologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Regulação para Cima , Via de Sinalização Wnt , beta Catenina/genéticaRESUMO
Dermal fibroblasts are required for structural integrity of the skin and for hair follicle development. Uniform Wnt signaling activity is present in dermal fibroblast precursors preceding hair follicle initiation, but the functional requirement of dermal Wnt signaling at early stages of skin differentiation and patterning remains largely uncharacterized. We show in mice that epidermal Wnt ligands are required for uniform dermal Wnt signaling/ß-catenin activity and regulate fibroblast cell proliferation and initiation of hair follicle placodes. In the absence of dermal Wnt signaling/ß-catenin activity, patterned upregulation of epidermal ß-catenin activity and Edar expression are absent. Conversely, forced activation of ß-catenin signaling leads to the formation of thickened dermis, enlarged epidermal placodes and dermal condensates that result in prematurely differentiated enlarged hair follicles. These data reveal functional roles for dermal Wnt signaling/ß-catenin in fibroblast proliferation and in the epidermal hair follicle initiation program.
Assuntos
Fibroblastos/citologia , Regulação da Expressão Gênica no Desenvolvimento , Folículo Piloso/citologia , Pele/metabolismo , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Animais , Proliferação de Células , Sobrevivência Celular , Ligantes , Camundongos , Estrutura Terciária de Proteína , Transdução de Sinais , Pele/embriologia , Células-Tronco/citologia , Fatores de TempoRESUMO
The bones of the mammalian skull vault form through intramembranous ossification. Skull bones ossify directly, in a process regulated by ß-catenin, instead of passing through a cartilage intermediate. We tested whether ß-catenin is necessary for fate selection of intramembranous bone progenitors in the skull. Here, we show in mice that removal of ß-catenin from skull bone progenitors results in the near complete transformation of the skull bones to cartilage, whereas constitutive ß-catenin activation inhibits skull bone fate selection. ß-catenin directly activated Twist1 expression in skull progenitors, conditional Twist1 deletion partially phenocopied the absence of ß-catenin, and Twist1 deletion partially restored bone formation in the presence of constitutive ß-catenin activation. Finally, Twist1 bound robustly to the 3'UTR of Sox9, the central initiator of chondrogenesis, suggesting that Twist1 might directly repress cartilage formation through Sox9. These findings provide insight into how ß-catenin signaling via Twist1 actively suppresses the formation of cartilage and promotes intramembranous ossification in the skull.
Assuntos
Condrogênese , Proteínas Nucleares/metabolismo , Crânio/citologia , Crânio/embriologia , Células-Tronco/fisiologia , Proteína 1 Relacionada a Twist/metabolismo , beta Catenina/metabolismo , Animais , Desenvolvimento Ósseo , Cartilagem/citologia , Diferenciação Celular , Linhagem Celular , Linhagem da Célula , Camundongos , Camundongos Endogâmicos C3H , Camundongos Transgênicos , Proteínas Nucleares/genética , Osteoblastos/metabolismo , Osteogênese , Regiões Promotoras Genéticas , Fatores de Transcrição SOX9/metabolismo , Transdução de Sinais , Proteína 1 Relacionada a Twist/genéticaRESUMO
Skin fibrosis is characterized by fibroblast activation and intradermal fat loss, resulting in excess deposition and remodeling of dermal extracellular matrix (ECM). The topography of the dominant ECM proteins, such as collagens, can indicate skin stiffness and remains understudied in evaluating fibrotic skin. Here, we adapted two different unbiased image analysis algorithms to define collagen topography and alignment in a genetically inducible and reversible Wnt activation fibrosis model. We demonstrated that Wnt-activated fibrotic skin has altered collagen fiber characteristics and a loss of collagen alignment, which were restored in the reversible model. This study highlights how unbiased algorithms can be used to analyze ECM topography, providing novel avenues to evaluate fibrotic skin onset, recovery, and treatment.
RESUMO
Cranial dermis develops from cephalic mesoderm and neural crest cells, but what signal(s) specifies the dermal lineage is unclear. Using genetic tools to fate map and manipulate a cranial mesenchymal progenitor population in the supraorbital region, we show that the dermal progenitor cells beneath the surface ectoderm process canonical Wnt signaling at the time of specification. We show that Wnt signaling/ß-catenin is absolutely required and sufficient for Dermo1 expression and dermal cell identity in the cranium. The absence of the Wnt signaling cue leads to formation of cartilage in craniofacial and ventral trunk regions at the expense of dermal and bone lineages. Dermo1 can be a direct transcription target and may mediate the functional role of Wnt signaling in dermal precursors. This study reveals a lineage-specific role of canonical Wnt signaling/ß-catenin in promoting dermal cell fate in distinct precursor populations.