RESUMEN
The conserved bazooka (baz/par3) gene acts as a key regulator of asymmetrical cell divisions across the animal kingdom. Associated Par3/Baz-Par6-aPKC protein complexes are also well known for their role in the establishment of apical/basal cell polarity in epithelial cells. Here we define a novel, positive function of Baz/Par3 in the Notch pathway. Using Drosophila wing and eye development, we demonstrate that Baz is required for Notch signaling activity and optimal transcriptional activation of Notch target genes. Baz appears to act independently of aPKC in these contexts, as knockdown of aPKC does not cause Notch loss-of-function phenotypes. Using transgenic Notch constructs, our data positions Baz activity downstream of activating Notch cleavage steps and upstream of Su(H)/CSL transcription factor complex activity on Notch target genes. We demonstrate a biochemical interaction between NICD and Baz, suggesting that Baz is required for NICD activity before NICD binds to Su(H). Taken together, our data define a novel role of the polarity protein Baz/Par3, as a positive and direct regulator of Notch signaling through its interaction with NICD.
Asunto(s)
Proteínas de Drosophila , Receptores Notch , Transducción de Señal , Alas de Animales , Animales , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Receptores Notch/metabolismo , Alas de Animales/metabolismo , Alas de Animales/embriología , Alas de Animales/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Unión Proteica , Drosophila melanogaster/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/genética , Ojo/embriología , Ojo/metabolismo , Ojo/crecimiento & desarrollo , Drosophila/metabolismo , Drosophila/embriología , Polaridad Celular , Péptidos y Proteínas de Señalización IntracelularRESUMEN
The acquisition of wings was a key event in insect evolution. As hemimetabolous insects were the first group to acquire functional wings, establishing the mechanisms of wing formation in this group could provide useful insights into their evolution. In this study, we aimed to elucidate the expression and function of the gene scalloped (sd), which is involved in wing formation in Drosophila melanogaster, and in Gryllus bimaculatus mainly during postembryonic development. Expression analysis showed that sd is expressed in the tergal edge, legs, antennae, labrum, and cerci during embryogenesis and in the distal margin of the wing pads from at least the sixth instar in the mid to late stages. Because sd knockout caused early lethality, nymphal RNA interference experiments were performed. Malformations were observed in the wings, ovipositor, and antennae. By analyzing the effects on wing morphology, it was revealed that sd is mainly involved in the formation of the margin, possibly through the regulation of cell proliferation. In conclusion, sd might regulate the local growth of wing pads and influence wing margin morphology in Gryllus.
Asunto(s)
Desarrollo Embrionario , Gryllidae , Proteínas de Insectos , Factores de Transcripción , Alas de Animales , Animales , Ciclo Celular , Proliferación Celular , Desarrollo Embrionario/genética , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Alas de Animales/embriología , Alas de Animales/metabolismo , Gryllidae/embriología , Gryllidae/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Morphogens of the Hh family trigger gene expression changes in receiving cells in a concentration-dependent manner to regulate their identity, proliferation, death or metabolism, depending on the tissue or organ. This variety of responses relies on a conserved signaling pathway. Its logic includes a negative-feedback loop involving the Hh receptor Ptc. Here, using experiments and computational models we study and compare the different spatial signaling profiles downstream of Hh in several developing Drosophila organs. We show that the spatial distributions of Ptc and the activator transcription factor CiA in wing, antenna and ocellus show similar features, but are markedly different from that in the compound eye. We propose that these two profile types represent two time points along the signaling dynamics, and that the interplay between the spatial displacement of the Hh source in the compound eye and the negative-feedback loop maintains the receiving cells effectively in an earlier stage of signaling. These results show how the interaction between spatial and temporal dynamics of signaling and differentiation processes may contribute to the informational versatility of the conserved Hh signaling pathway.
Asunto(s)
Drosophila , Proteínas Hedgehog , Transducción de Señal , Drosophila/embriología , Animales , Proteínas Hedgehog/fisiología , Alas de Animales/embriología , Ojo Compuesto de los Artrópodos/embriologíaRESUMEN
A fundamental question in biology is how embryonic development is timed between different species. To address this problem, we compared wing development in the quail and the larger chick. We reveal that pattern formation is faster in the quail as determined by the earlier activation of 5'Hox genes, termination of developmental organizers (Shh and Fgf8), and the laying down of the skeleton (Sox9). Using interspecies tissue grafts, we show that developmental timing can be reset during a critical window of retinoic acid signaling. Accordingly, extending the duration of retinoic acid signaling switches developmental timing between the quail and the chick and the chick and the larger turkey. However, the incremental growth rate is comparable between all three species, suggesting that the pace of development primarily governs differences in the expansion of the skeletal pattern. The widespread distribution of retinoic acid could coordinate developmental timing throughout the embryo.
Asunto(s)
Desarrollo Embrionario/fisiología , Inducción Embrionaria/fisiología , Tretinoina/metabolismo , Alas de Animales/embriología , Animales , Embrión de Pollo , Codorniz/embriología , Pavos/embriologíaRESUMEN
A long-standing view in the field of evo-devo is that insect forewings develop without any Hox gene input. The Hox gene Antennapedia (Antp), despite being expressed in the thoracic segments of insects, has no effect on wing development. This view has been obtained from studies in two main model species: Drosophila and Tribolium. Here, we show that partial loss of function of Antp resulted in reduced and malformed adult wings in Bombyx, Drosophila and Tribolium. Antp mediates wing growth in Bombyx by directly regulating the ecdysteriod biosynthesis enzyme gene (shade) in the wing tissue, which leads to local production of the growth hormone 20-hydroxyecdysone. Additional targets of Antp are wing cuticular protein genes CPG24, CPH28 and CPG9, which are essential for wing development. We propose, therefore, that insect wing development occurs in an Antp-dependent manner. This article has an associated 'The people behind the papers' interview.
Asunto(s)
Proteínas de Homeodominio/metabolismo , Proteínas de Insectos/metabolismo , Alas de Animales/embriología , Animales , Bombyx , Drosophila , Ecdisterona/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Insectos/genética , Mutación con Pérdida de Función , Morfogénesis , Tribolium , Alas de Animales/metabolismoRESUMEN
Regeneration is a response mechanism aiming to reconstruct lost or damaged structures. To achieve this, the cells repopulating the lost tissue often have to change their original identity, a process that involves chromatin remodelling.We have analysed the issue of chromatin remodelling during regeneration in the wing disc of Drosophila . In this disc the ablation of the central region (the pouch) induces the regenerative response of the cells from the lateral region (the hinge), which reconstitute the wing pouch. We have examined euchromatin and heterochromatin histone marks during the process and find that heterochromatin marks disappear but are recovered when regeneration is complete. Euchromatin marks are not modified. We also describe the transcription of two retrotransposons, Roo and F-element in the regenerating cells. We have established a temporal correlation between the alterations of heterochromatin marks and the levels of transcription of two retrotransposons, Roo and F-element, both during embryonic development and in the regeneration process.
Asunto(s)
Ensamble y Desensamble de Cromatina/genética , Drosophila melanogaster/embriología , Discos Imaginales/crecimiento & desarrollo , Regeneración/fisiología , Retroelementos/genética , Alas de Animales/embriología , Acetilación , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/fisiología , Eucromatina/metabolismo , Heterocromatina/metabolismo , Histonas/metabolismo , Elementos de Nucleótido Esparcido Largo/genética , Metilación , Alas de Animales/crecimiento & desarrolloRESUMEN
Spatial boundaries formed during animal development originate from the pre-patterning of tissues by signaling molecules, called morphogens. The accuracy of boundary location is limited by the fluctuations of morphogen concentration that thresholds the expression level of target gene. Producing more morphogen molecules, which gives rise to smaller relative fluctuations, would better serve to shape more precise target boundaries; however, it incurs more thermodynamic cost. In the classical diffusion-depletion model of morphogen profile formation, the morphogen molecules synthesized from a local source display an exponentially decaying concentration profile with a characteristic length λ. Our theory suggests that in order to attain a precise profile with the minimal cost, λ should be roughly half the distance to the target boundary position from the source. Remarkably, we find that the profiles of morphogens that pattern the Drosophila embryo and wing imaginal disk are formed with nearly optimal λ. Our finding underscores the cost-effectiveness of precise morphogen profile formation in Drosophila development.
Asunto(s)
Tipificación del Cuerpo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Modelos Biológicos , Animales , Difusión , Proteínas de Drosophila/genética , Drosophila melanogaster/embriología , Drosophila melanogaster/genética , Embrión no Mamífero/metabolismo , Regulación del Desarrollo de la Expresión Génica , Discos Imaginales/metabolismo , Transducción de Señal , Factores de Tiempo , Alas de Animales/embriología , Alas de Animales/metabolismoRESUMEN
Morphogen signaling proteins disperse across tissues to activate signal transduction in target cells. We investigated dispersion of Hedgehog (Hh), Wnt homolog Wingless (Wg), and Bone morphogenic protein homolog Decapentaplegic (Dpp) in the Drosophila wing imaginal disc. We discovered that delivery of Hh, Wg, and Dpp to their respective targets is regulated. We found that <5% of Hh and <25% of Wg are taken up by disc cells and activate signaling. The amount of morphogen that is taken up and initiates signaling did not change when the level of morphogen expression was varied between 50 and 200% (Hh) or 50 and 350% (Wg). Similar properties were observed for Dpp. We analyzed an area of 150 µm×150 µm that includes Hh-responding cells of the disc as well as overlying tracheal cells and myoblasts that are also activated by disc-produced Hh. We found that the extent of signaling in the disc was unaffected by the presence or absence of the tracheal and myoblast cells, suggesting that the mechanism that disperses Hh specifies its destinations to particular cells, and that target cells do not take up Hh from a common pool.
Asunto(s)
Proteínas de Drosophila/metabolismo , Proteínas Hedgehog/metabolismo , Discos Imaginales/metabolismo , Transducción de Señal , Proteína Wnt1/metabolismo , Animales , Drosophila/metabolismo , Proteínas de Drosophila/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas Hedgehog/genética , Morfogénesis , Alas de Animales/embriología , Proteína Wnt1/genéticaRESUMEN
The Notch signaling pathway is highly conserved and regulates various fundamental development events. Activation of Notch signaling relies on production of the Notch intracellular domain (NICD), which assembles a transcription factor complex to turn on down-stream targets expression. The mastermind (mam) gene encodes an essential co-activator that permits NICD activity in the cell nucleus. During a somatic mosaic screen in Drosophila, an uncharacterized gene l(2)S9998 is identified as a positive regulator of the Notch signaling pathway. Genetic analysis demonstrates that l(2)S9998 functions at the level of transcriptional activation of Notch targets in the signal receiving cells. Whole genome sequencing reveals that l(2)S9998 is a novel allele of the mam gene, which is further confirmed by complementation tests. Along with three molecularly defined transposon insertions isolated from the screen, four mutants of mam are shown to modulate Notch signaling during fly wing development. Our analysis provides additional genetic resources for understanding mam function and Notch signaling regulation.
Asunto(s)
Alelos , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Pruebas Genéticas , Proteínas Nucleares/genética , Animales , Mapeo Cromosómico , Elementos Transponibles de ADN/genética , Mutagénesis Insercional/genética , Receptores Notch/metabolismo , Transducción de Señal , Alas de Animales/embriologíaRESUMEN
Tissue organization is often characterized by specific patterns of cell morphology. How such patterns emerge in developing tissues is a fundamental open question. Here, we investigate the emergence of tissue-scale patterns of cell shape and mechanical tissue stress in the Drosophila wing imaginal disc during larval development. Using quantitative analysis of the cellular dynamics, we reveal a pattern of radially oriented cell rearrangements that is coupled to the buildup of tangential cell elongation. Developing a laser ablation method, we map tissue stresses and extract key parameters of tissue mechanics. We present a continuum theory showing that this pattern of cell morphology and tissue stress can arise via self-organization of a mechanical feedback that couples cell polarity to active cell rearrangements. The predictions of this model are supported by knockdown of MyoVI, a component of mechanosensitive feedback. Our work reveals a mechanism for the emergence of cellular patterns in morphogenesis.
During development, carefully choreographed cell movements ensure the creation of a healthy organism. To determine their identity and place across a tissue, cells can read gradients of far-reaching signaling molecules called morphogens; in addition, physical forces can play a part in helping cells acquire the right size and shape. Indeed, cells are tightly attached to their neighbors through connections linked to internal components. Structures or proteins inside the cells can pull on these junctions to generate forces that change the physical features of a cell. However, it is poorly understood how these forces create patterns of cell size and shape across a tissue. Here, Dye, Popovic et al. combined experiments with physical models to examine how cells acquire these physical characteristics across the developing wing of fruit fly larvae. This revealed that cells pushing and pulling on one another create forces that trigger internal biochemical reorganization for instance, force-generating structures become asymmetrical. In turn, the cells exert additional forces on their neighbors, setting up a positive feedback loop which results in cells adopting the right size and shape across the organ. As such, cells in the fly wing can spontaneously self-organize through the interplay of mechanical and biochemical signals, without the need for pre-existing morphogen gradients. A refined understanding of how physical forces shape cells and organs would help to grasp how defects can emerge during development. This knowledge would also allow scientists to better grow tissues and organs in the laboratory, both for theoretical research and regenerative medicine.
Asunto(s)
Forma de la Célula , Drosophila melanogaster/fisiología , Discos Imaginales/fisiología , Mecanotransducción Celular , Alas de Animales/fisiología , Animales , Tipificación del Cuerpo , División Celular , Polaridad Celular , Drosophila melanogaster/embriología , Drosophila melanogaster/genética , Retroalimentación Fisiológica , Femenino , Discos Imaginales/embriología , Masculino , Modelos Biológicos , Cadenas Pesadas de Miosina/genética , Cadenas Pesadas de Miosina/metabolismo , Estrés Mecánico , Factores de Tiempo , Alas de Animales/embriologíaRESUMEN
Regeneration is a common phenomenon in various organisms by which tissues restore the damaged or naturally detached parts. In insects, appendage regeneration takes place during the embryonic, larval and pupal stages for individual survival. The wing disc of black cutworm Agrotis ypsilon has the capacity of regeneration after ablation, but understanding of molecular mechanisms in wing disc regeneration is still limited. After ablation of partial or whole wing discs before the fifth instar larval stage, the adult wings appeared to be normal. In the last two larval stages, ablation of the left wing disc led to smaller corresponding adult wing. Cell proliferation was reduced in the ablated wing disc but was gradually recovered two days post ablation. Transcriptome analysis found that genes in the mitogen-activated protein kinase (MAPK) pathway were upregulated. Repression of gene expression in this pathway, including Ras oncogene at 64B (Ras64B), Downstream of raf1 (Dsor1), and cAMP-dependent protein kinase catalytic subunit 3 (Pka-C3) by RNA interference after ablation, led to diminishment of both adult wings, suggesting that the MAPK signaling is essential for wing growth. Additionally, cell proliferation was still decelerated by injecting Ras64B, Dsor, or Pka-C3 dsRNA two days after ablation, indicating that the MAPK signaling-regulated cell proliferation is essential for growth. These results provide molecular clues to the regulation of cell proliferation during regeneration in lepidopteran insects.
Asunto(s)
Mariposas Nocturnas , Regeneración , Alas de Animales , Animales , Proliferación Celular/genética , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes de Insecto , Genes ras , Larva/metabolismo , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Mariposas Nocturnas/embriología , Mariposas Nocturnas/metabolismo , Pupa/metabolismo , Regeneración/genética , Regeneración/fisiología , Transducción de Señal , Alas de Animales/embriología , Alas de Animales/crecimiento & desarrolloRESUMEN
Matrix metalloproteinases (MMPs) are the major proteinases that process or degrade numerous extracellular matrix (ECM) components and are evolutionarily conserved from nematodes to humans. During molting in insects, the old cuticle is removed and replaced by a new counterpart. Although the regulatory mechanisms of hormones and nutrients in molting have been well studied, very little is known about the roles of ECM-modifying enzymes in this process. Here, we found that MMPs are necessary for imaginal molting of the American cockroach, Periplaneta americana. Inhibition of Mmp activity via inhibitor treatment led to the failure of eclosion and wing expansion. Five Mmps genes were identified from the P. americana genome, and PaMmp2 played the dominant roles during molting. Further microscopic investigations showed that newly formed adult cuticles were attenuated and that then chitin content was reduced upon Mmp inhibition. Transcriptomic analysis of the integument demonstrated that multiple signaling and metabolic pathways were changed. Microscopic investigation of the wings showed that epithelial cells were restrained together because they were incapable of degrading the ECM upon Mmp inhibition. Transcriptomic analysis of the wing identified dozens of possible genes functioned in wing expansion. This is the first study to show the essential roles of Mmps in the nymph-adult transition of hemimetabolous insects.
Asunto(s)
Metaloproteinasas de la Matriz , Periplaneta , Alas de Animales , Animales , Quitina/metabolismo , Perfilación de la Expresión Génica , Genes de Insecto , Larva/metabolismo , Metaloproteinasas de la Matriz/genética , Metaloproteinasas de la Matriz/metabolismo , Metamorfosis Biológica , Muda , Ninfa/metabolismo , Periplaneta/embriología , Periplaneta/genética , Periplaneta/metabolismo , Periplaneta/fisiología , Alas de Animales/embriología , Alas de Animales/metabolismoRESUMEN
Chitinase degrades chitin in the old epidermis or peritrophic matrix of insects, which ensures normal development and metamorphosis. In our previous work, we comprehensively studied the function of SfCht7 in Sogatella furcifera. However, the number and function of chitinase genes in S. furcifera remain unknown. Here, we identified 12 full-length chitinase transcripts from S. furcifera, which included nine chitinase (Cht), two imaginal disc growth factor (IDGF), and one endo-ß-N-acetylglucosaminidase (ENGase) genes. Expression analysis results revealed that the expression levels of eight genes (SfCht3, SfCht5, SfCht6-1, SfCht6-2, SfCht7, SfCht8, SfCht10, and SfIDGF2) with similar transcript levels peaked prior to molting of each nymph and were highly expressed in the integument. Based on RNA interference (RNAi), description of the functions of each chitinase gene indicated that the silencing of SfCht5, SfCht10, and SfIDGF2 led to molting defects and lethality. RNAi inhibited the expressions of SfCht5, SfCht7, SfCht10, and SfIDGF2, which led to downregulated expressions of chitin synthase 1 (SfCHS1, SfCHS1a, and SfCHS1b) and four chitin deacetylase genes (SfCDA1, SfCDA2, SfCDA3, and SfCDA4), and caused a change in the expression level of two trehalase genes (TRE1 and TRE2). Furthermore, silencing of SfCht7 induced a significant decrease in the expression levels of three wing development-related genes (SfWG, SfDpp, and SfHh). In conclusion, SfCht5, SfCht7, SfCht10, and SfIDGF2 play vital roles in nymph-adult transition and are involved in the regulation of chitin metabolism, and SfCht7 is also involved in wing development; therefore, these genes are potential targets for control of S. furcifera.
Asunto(s)
Quitinasas/genética , Hemípteros , Metamorfosis Biológica/genética , Acetilglucosaminidasa/genética , Exoesqueleto/embriología , Exoesqueleto/crecimiento & desarrollo , Animales , Regulación del Desarrollo de la Expresión Génica , Genes de Insecto , Hemípteros/embriología , Hemípteros/genética , Hemípteros/fisiología , Discos Imaginales/embriología , Péptidos y Proteínas de Señalización Intercelular/genética , Muda/genética , Ninfa/crecimiento & desarrollo , Ninfa/fisiología , Alas de Animales/embriología , Alas de Animales/crecimiento & desarrolloRESUMEN
Cell differentiation, proliferation, and morphogenesis are generally driven by instructive signals that are sent and interpreted by adjacent tissues, a process known as induction. Cell recruitment is a particular case of induction in which differentiated cells produce a signal that drives adjacent cells to differentiate into the same type as the inducers. Once recruited, these new cells may become inducers to continue the recruitment process, closing a feed-forward loop that propagates the growth of a specific cell-type population. So far, little attention has been given to cell recruitment as a developmental mechanism. Here, we review the components of cell recruitment and discuss its contribution to development in three different examples: the Drosophila wing, the vertebrate inner ear, and the mammalian thyroid gland. Finally, we posit some open questions about the role of cell recruitment in organ patterning and growth.
Asunto(s)
Drosophila , Mamíferos , Morfogénesis , Vertebrados , Animales , Drosophila/embriología , Oído Interno/embriología , Regulación del Desarrollo de la Expresión Génica , Mamíferos/embriología , Glándula Tiroides/embriología , Vertebrados/embriología , Alas de Animales/embriologíaRESUMEN
Cell extrusion is a crucial regulator of epithelial tissue development and homeostasis. Epithelial cells undergoing apoptosis, bearing pathological mutations or possessing developmental defects are actively extruded toward elimination. However, the molecular mechanisms of Drosophila epithelial cell extrusion are not fully understood. Here, we report that activation of the conserved Hippo (Hpo) signaling pathway induces both apical and basal cell extrusion in the Drosophila wing disc epithelia. We show that canonical Yorkie targets Diap1, Myc and Cyclin E are not required for either apical or basal cell extrusion induced by activation of this pathway. Another target gene, bantam, is only involved in basal cell extrusion, suggesting novel Hpo-regulated apical cell extrusion mechanisms. Using RNA-seq analysis, we found that JNK signaling is activated in the extruding cells. We provide genetic evidence that JNK signaling activation is both sufficient and necessary for Hpo-regulated cell extrusion. Furthermore, we demonstrate that the ETS-domain transcription factor Ets21c, an ortholog of proto-oncogenes FLI1 and ERG, acts downstream of JNK signaling to mediate apical cell extrusion. Our findings reveal a novel molecular link between Hpo signaling and cell extrusion.
Asunto(s)
Proteínas de Drosophila/metabolismo , Discos Imaginales/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-ets/metabolismo , Transducción de Señal/fisiología , Alas de Animales/embriología , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster , Discos Imaginales/citología , Proteínas Inhibidoras de la Apoptosis/genética , Proteínas Inhibidoras de la Apoptosis/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , MAP Quinasa Quinasa 4/genética , MAP Quinasa Quinasa 4/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Proteínas Serina-Treonina Quinasas/genética , Proteínas Proto-Oncogénicas c-ets/genética , Transactivadores/genética , Transactivadores/metabolismo , Alas de Animales/citología , Proteínas Señalizadoras YAPRESUMEN
Hedgehog (Hh) is an evolutionarily conserved signaling protein that has essential roles in animal development and homeostasis. We investigated Hh signaling in the region of the Drosophila wing imaginal disc that produces Hh and is near the tracheal air sac primordium (ASP) and myoblasts. Hh distributes in concentration gradients in the anterior compartment of the wing disc, ASP and myoblasts, and activates genes in each tissue. Some targets of Hh signal transduction are common to the disc, ASP and myoblasts, whereas others are tissue-specific. Signaling in the three tissues is cytoneme-mediated and cytoneme-dependent. Some ASP cells project cytonemes that receive both Hh and Branchless (Bnl), and some targets regulated by Hh signaling in the ASP are also dependent on Bnl signal transduction. We conclude that the single source of Hh in the wing disc regulates cell type-specific responses in three discreet target tissues.
Asunto(s)
Proteínas de Drosophila/metabolismo , Proteínas Hedgehog/metabolismo , Discos Imaginales/metabolismo , Transducción de Señal , Alas de Animales/embriología , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster , Factores de Crecimiento de Fibroblastos/genética , Factores de Crecimiento de Fibroblastos/metabolismo , Proteínas Hedgehog/genética , Discos Imaginales/citología , Alas de Animales/citologíaRESUMEN
Epithelial sheets define organ architecture during development. Here, we employed an iterative multiscale computational modeling and quantitative experimental approach to decouple direct and indirect effects of actomyosin-generated forces, nuclear positioning, extracellular matrix, and cell-cell adhesion in shaping Drosophila wing imaginal discs. Basally generated actomyosin forces generate epithelial bending of the wing disc pouch. Surprisingly, acute pharmacological inhibition of ROCK-driven actomyosin contractility does not impact the maintenance of tissue height or curved shape. Computational simulations show that ECM tautness provides only a minor contribution to modulating tissue shape. Instead, passive ECM pre-strain serves to maintain the shape independent from actomyosin contractility. These results provide general insight into how the subcellular forces are generated and maintained within individual cells to induce tissue curvature. Thus, the results suggest an important design principle of separable contributions from ECM prestrain and actomyosin tension during epithelial organogenesis and homeostasis.
Asunto(s)
Actomiosina/metabolismo , Epitelio/anatomía & histología , Matriz Extracelular/metabolismo , Animales , Drosophila/anatomía & histología , Drosophila/embriología , Drosophila/metabolismo , Epitelio/metabolismo , Fosforilación , Alas de Animales/anatomía & histología , Alas de Animales/embriología , Alas de Animales/metabolismoRESUMEN
Differential specification of dorsal flight appendages, wing and haltere, in Drosophila provides an excellent model system to address a number of important questions in developmental biology at the levels of molecules, pathways, tissues, organs, organisms and evolution. Here we discuss the mechanism by which the Hox protein Ubx recognizes and regulates its downstream targets, implications of the same in growth control at cellular and organ level and finally the evolution of haltere from ancestral hindwings in other holometabolous insects.
Asunto(s)
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/fisiología , Proteínas de Homeodominio/metabolismo , Organogénesis , Factores de Transcripción/metabolismo , Alas de Animales/embriología , Alas de Animales/fisiología , Animales , Proteínas de Drosophila/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Factores de Transcripción/genética , Alas de Animales/anatomía & histologíaRESUMEN
Morphogens are important signalling molecules for tissue development and their secretion requires tight regulation. In the wing imaginal disc of flies, the morphogen Wnt/Wingless is apically presented by the secreting cell and re-internalized before final long-range secretion. Why Wnt molecules undergo these trafficking steps and the nature of the regulatory control within the endosomal compartment remain unclear. Here, we have investigated how Wnts are sorted at the level of endosomes by the versatile v-SNARE Ykt6. Using in vivo genetics, proximity-dependent proteomics and in vitro biochemical analyses, we show that most Ykt6 is present in the cytosol, but can be recruited to de-acidified compartments and recycle Wnts to the plasma membrane via Rab4-positive recycling endosomes. Thus, we propose a molecular mechanism by which producing cells integrate and leverage endocytosis and recycling via Ykt6 to coordinate extracellular Wnt levels.
Asunto(s)
Proteínas de Drosophila/metabolismo , Endosomas/metabolismo , Proteínas R-SNARE/metabolismo , Alas de Animales/embriología , Proteínas Wnt/metabolismo , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster , Endosomas/genética , Epitelio/embriología , Proteínas R-SNARE/genética , Proteínas Wnt/genéticaRESUMEN
While rare pathogenic copy-number variants (CNVs) are associated with both neuronal and non-neuronal phenotypes, functional studies evaluating these regions have focused on the molecular basis of neuronal defects. We report a systematic functional analysis of non-neuronal defects for homologs of 59 genes within ten pathogenic CNVs and 20 neurodevelopmental genes in Drosophila melanogaster. Using wing-specific knockdown of 136 RNA interference lines, we identified qualitative and quantitative phenotypes in 72/79 homologs, including 21 lines with severe wing defects and six lines with lethality. In fact, we found that 10/31 homologs of CNV genes also showed complete or partial lethality at larval or pupal stages with ubiquitous knockdown. Comparisons between eye and wing-specific knockdown of 37/45 homologs showed both neuronal and non-neuronal defects, but with no correlation in the severity of defects. We further observed disruptions in cell proliferation and apoptosis in larval wing discs for 23/27 homologs, and altered Wnt, Hedgehog and Notch signaling for 9/14 homologs, including AATF/Aatf, PPP4C/Pp4-19C, and KIF11/Klp61F. These findings were further supported by tissue-specific differences in expression patterns of human CNV genes, as well as connectivity of CNV genes to signaling pathway genes in brain, heart and kidney-specific networks. Our findings suggest that multiple genes within each CNV differentially affect both global and tissue-specific developmental processes within conserved pathways, and that their roles are not restricted to neuronal functions.