RESUMEN
Establishment of the dorsal-ventral (DV) axis of the Drosophila embryo depends on ventral activation of the maternal Toll pathway, which creates a gradient of the NFkB/c-rel-related transcription factor dorsal. Signaling through the maternal BMP pathway also alters the dorsal gradient, probably by regulating degradation of the IkB homologue Cactus. The BMP4 homologue decapentaplegic (dpp) and the BMP antagonist short gastrulation (sog) are expressed by follicle cells during mid-oogenesis, but it is unknown how they affect embryonic patterning following fertilization. Here, we provide evidence that maternal Sog and Dpp proteins are secreted into the perivitelline space where they remain until early embryogenesis to modulate Cactus degradation, enabling their dual function in patterning the eggshell and embryo. We find that metalloproteases encoded by tolloid (tld) and tolkin (tok), which cleave Sog, are expressed by follicle cells and are required to generate DV asymmetry in the Dpp signal. Expression of tld and tok is ventrally restricted by the TGF-alpha ligand encoded by gurken, suggesting that signaling via the EGF receptor pathway may regulate embryonic patterning through two independent mechanisms: by restricting the expression of pipe and thereby activation of Toll signaling and by spatially regulating BMP activity.
Asunto(s)
Tipificación del Cuerpo/fisiología , Proteínas de Drosophila/fisiología , Inducción Embrionaria/fisiología , Animales , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/biosíntesis , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Proteínas del Huevo/biosíntesis , Proteínas del Huevo/fisiología , Óvulo/metabolismo , Fosfoproteínas/metabolismoRESUMEN
We performed a systematic analysis of 929 human disease gene entries associated with at least one mutant allele in the Online Mendelian Inheritance in Man (OMIM) database against the recently completed genome sequence of Drosophila melanogaster. The results of this search have been formatted as an updateable and searchable on-line database called Homophila. Our analysis identified 714 distinct human disease genes (77% of disease genes searched) matching 548 unique Drosophila sequences, which we have summarized by disease category. This breakdown into disease classes creates a picture of disease genes that are amenable to study using Drosophila as the model organism. Of the 548 Drosophila genes related to human disease genes, 153 are associated with known mutant alleles and 56 more are tagged by P-element insertions in or near the gene. Examples of how to use the database to identify Drosophila genes related to human disease genes are presented. We anticipate that cross-genomic analysis of human disease genes using the power of Drosophila second-site modifier screens will promote interaction between human and Drosophila research groups, accelerating the understanding of the pathogenesis of human genetic disease. The Homophila database is available at http://homophila.sdsc.edu.
Asunto(s)
Drosophila melanogaster/genética , Genes de Insecto/genética , Enfermedades Genéticas Congénitas/genética , Secuencia de Aminoácidos/genética , Animales , Biología Computacional/métodos , Secuencia Conservada/genética , Bases de Datos Factuales , Humanos , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico , Transducción de Señal/genéticaRESUMEN
The Drosophila rhomboid (rho) gene participates in localized activation of EGF-receptor signaling in various developmental settings. The Rhomboid protein has been proposed to promote presentation and/or processing of the membrane-bound Spitz (mSpi) EGF-related ligand to generate an active diffusible form of the ligand. Here, we report on a new rhomboid-related gene identified by sequence similarity searching that we have named brother of rhomboid (brho). In contrast to rho, which is expressed in complex patterns during many stages of development, brho appears to be expressed only during oogenesis. brho transcripts are present in early oocytes and abut posterior follicle cells which exhibit high levels of MAPK activation. brho, like rho, collaborates with Star to promote signaling through the EGF-R/MAPK pathway, and genetic evidence indicates that Brho can activate both the mSpi and the Grk precursor EGF ligands in the wing. We propose that endogenous brho may activate the oocyte-specific Gurken ligand and thereby participate in defining posterior cell fates in the early follicular epithelium.
Asunto(s)
Proteínas de Drosophila , Drosophila melanogaster/genética , Factor de Crecimiento Epidérmico , Receptores ErbB/fisiología , Regulación del Desarrollo de la Expresión Génica , Proteínas de Insectos/genética , Sistema de Señalización de MAP Quinasas/genética , Oogénesis/genética , Factor de Crecimiento Transformador alfa , Secuencia de Aminoácidos , Animales , Drosophila melanogaster/embriología , Drosophila melanogaster/crecimiento & desarrollo , Embrión no Mamífero/metabolismo , Activación Enzimática , Femenino , Proteínas de Insectos/biosíntesis , Proteínas de Insectos/fisiología , Larva/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/fisiología , Datos de Secuencia Molecular , Familia de Multigenes , Oocitos/metabolismo , Oogénesis/fisiología , Folículo Ovárico/citología , Folículo Ovárico/metabolismo , Fosfoproteínas/fisiología , Filogenia , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Transcripción Genética , Factores de Crecimiento Transformadores/fisiología , Alas de Animales/embriologíaRESUMEN
It has been proposed that wing veins in Drosophila form at boundaries between discrete sectors of cells that subdivide the anterior-posterior axis of the developing wing primordium. Recently, analysis of events underlying initiation of vein formation suggests that there is a general developmental mechanism for drawing lines between adjacent domains of cells, which is referred to as 'for-export-only-signaling'. In this model, cells in one domain produce a short range signal to which they cannot respond. As a consequence of this constraint, cells lying in a narrow line immediately outside the signal-producing domain are the only cells that can respond to the signal by activating expression of vein-promoting genes.
Asunto(s)
Drosophila melanogaster/crecimiento & desarrollo , Alas de Animales/crecimiento & desarrollo , Animales , Evolución Biológica , Drosophila melanogaster/genéticaRESUMEN
The short gastrulation (sog) and decapentaplegic (dpp) genes function antagonistically in the early Drosophila zygote to pattern the dorsoventral (DV) axis of the embryo. This interplay between sog and dpp determines the extent of the neuroectoderm and subdivides the dorsal ectoderm into two territories. Here, we present evidence that sog and dpp also play opposing roles during oogenesis in patterning the DV axis of the embryo. We show that maternally produced Dpp increases levels of the I(kappa)B-related protein Cactus and reduces the magnitude of the nuclear concentration gradient of the NF(kappa)B-related Dorsal protein, and that Sog limits this effect. We present evidence suggesting that Dpp signaling increases Cactus levels by reducing a signal-independent component of Cactus degradation. Epistasis experiments reveal that sog and dpp act downstream of, or in parallel to, the Toll receptor to reduce translocation of Dorsal protein into the nucleus. These results broaden the role previously defined for sog and dpp in establishing the embryonic DV axis and reveal a novel form of crossregulation between the NF(kappa)B and TGF(beta) signaling pathways in pattern formation.
Asunto(s)
Tipificación del Cuerpo/fisiología , Proteínas de Drosophila , Drosophila/embriología , Proteínas de Insectos/metabolismo , Proteínas de Insectos/fisiología , Glicoproteínas de Membrana/metabolismo , Receptores de Superficie Celular , Transducción de Señal , Factores de Transcripción , Factor de Crecimiento Transformador alfa , Animales , Vértebra Cervical Axis/fisiología , Núcleo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Drosophila/genética , Drosophila/fisiología , Femenino , Células Germinativas , Proteínas de Insectos/genética , Masculino , Proteínas Nucleares/metabolismo , Oogénesis/fisiología , Óvulo , Fosfoproteínas/metabolismo , Receptores Toll-Like , Factores de Crecimiento Transformadores/metabolismoRESUMEN
Structurally unrelated neural inducers in vertebrate and invertebrate embryos have been proposed to function by binding to BMP4 or Dpp, respectively, and preventing these homologous signals from activating their receptor(s). In this study, we investigate the functions of various forms of the Drosophila Sog protein using the discriminating assay of Drosophila wing development. We find that misexpression of Drosophila Sog, or its vertebrate counterpart Chordin, generates a very limited vein-loss phenotype. This sog misexpression phenotype is very similar to that of viable mutants of glass-bottom boat (gbb), which encodes a BMP family member. Consistent with Sog selectively interfering with Gbb signaling, Sog can block the effect of misexpressing Gbb, but not Dpp in the wing. In contrast to the limited BMP inhibitory activity of Sog, we have identified carboxy-truncated forms of Sog, referred to as Supersog, which when misexpressed cause a broad range of dpp(-) mutant phenotypes. In line with its phenotypic effects, Supersog can block the effects of both misexpressing Dpp and Gbb in the wing. Vertebrate Noggin, on the other hand, acts as a general inhibitor of Dpp signaling, which can interfere with the effect of overexpressing Dpp, but not Gbb. We present evidence that Sog processing occurs in vivo and is biologically relevant. Overexpression of intact Sog in embryos and adult wing primordia leads to the developmentally regulated processing of Sog. This in vivo processing of Sog can be duplicated in vitro by treating Sog with a combination of the metalloprotease Tolloid (Tld) plus Twisted Gastrulation (Tsg), another extracellular factor involved in Dpp signaling. In accord with this result, coexpression of intact Sog and Tsg in developing wings generates a phenotype very similar to that of Supersog. Finally, we provide evidence that tsg functions in the embryo to generate a Supersog-like activity, since Supersog can partially rescue tsg(-) mutants. Consistent with this finding, sog(- )and tsg(-) mutants exhibit similar dorsal patterning defects during early gastrulation. These results indicate that differential processing of Sog generates a novel BMP inhibitory activity during development and, more generally, that BMP antagonists play distinct roles in regulating the quality as well as the magnitude of BMP signaling.
Asunto(s)
Proteínas Morfogenéticas Óseas/antagonistas & inhibidores , Proteínas de Drosophila , Glicoproteínas , Proteínas de Insectos/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Proteínas/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Animales , Proteínas Portadoras , Línea Celular , Drosophila/crecimiento & desarrollo , Proteínas de Insectos/genética , Metaloendopeptidasas/metabolismo , Mutagénesis , Fenotipo , Proteínas/genética , Transducción de Señal , Metaloproteinasas Similares a Tolloid , Alas de Animales/crecimiento & desarrollo , Xenopus/embriologíaRESUMEN
Hedgehog (Hh) plays an important role in Drosophila wing patterning by inducing expression of Dpp, which serves to organize the wing globally across the A-P axis. We show here how Hh signalling also plays a direct role in patterning the medial wing through the activation of the Hh-target gene, knot (kn). kn is expressed in Hh-responsive cells near the A-P compartment boundary, where its expression is dependent on fu, a component of Hh signalling. kn is required for the proper positioning of veins 3 and 4 and to prevent ectopic venation between them. Furthermore, the expansion anteriorly of the normal kn expression domain causes an associated anterior shift in the position of vein 3 in the resultant wing. Ectopic expression of kn elsewhere in the wing imaginal disc results in the failure to properly activate the vein initiation genes, rho and Dl. Expression of the gene encoding the EGF-receptor (EGFR), which is required for vein initiation and subsequent differentiation, is normally depressed in the 3-4 intervein region. This downregulation of EGFR in the medial portion of the imaginal disc is dependent on kn activity and ectopic expression of kn inactivates EGFR elsewhere in the wing primordium. We propose kn expression in Hh-responsive cells of the wing blade anlagen during the late third instar creates a zone of cells in the medial wing in which vein primordia cannot be induced. The primordia for veins 3 and 4 are laid down adjacent to the kn-imposed vein-free zone, presumably by a signalling factor (such as Vn) also synthesized in the medial region of the wing.
Asunto(s)
Proteínas de Drosophila , Drosophila melanogaster/crecimiento & desarrollo , Proteínas de Insectos/fisiología , Factores de Transcripción/fisiología , Alas de Animales/crecimiento & desarrollo , Animales , Drosophila melanogaster/genética , Drosophila melanogaster/fisiología , Expresión Génica , Regulación de la Expresión Génica , Genes de Insecto , Proteínas de Insectos/genética , Proteínas de la Membrana/genética , Proteínas Nucleares/genética , Fosfoproteínas/genética , Factores de Transcripción/genéticaRESUMEN
Genes of the ventrolateral group in Drosophila are dedicated to developmental regulation of Egfr signaling in multiple processes including wing vein development. Among these genes, Egfr encodes the Drosophila EGF-Receptor, spitz (spi) and vein (vn) encode EGF-related ligands, and rhomboid (rho) and Star (S) encode membrane proteins. In this study, we show that rho-mediated hyperactivation of the EGFR/MAPK pathway is required for vein formation throughout late larval and early pupal development. Consistent with this observation, rho activity is necessary and sufficient to activate MAPK in vein primordium during late larval and early pupal stages. Epistasis studies using a dominant negative version of Egfr and a ligand-independent activated form of Egfr suggest that rho acts upstream of the receptor. We show that rho and S function in a common aspect of vein development since loss-of-function clones of rho or S result in nearly identical non-autonomous loss-of-vein phenotypes. Furthermore, mis-expression of rho and S in wild-type and mutant backgrounds reveals that these genes function in a synergistic and co-dependent manner. In contrast, spi does not play an essential role in the wing. These data indicate that rho and S act in concert, but independently of spi, to promote vein development through the EGFR/MAPK signaling pathway.
Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Proteínas de Drosophila , Drosophila/genética , Factor de Crecimiento Epidérmico , Receptores ErbB/metabolismo , Proteínas de la Membrana/genética , Neurregulinas , Alas de Animales/crecimiento & desarrollo , Animales , Proteínas Quinasas Dependientes de Calcio-Calmodulina/genética , Drosophila/crecimiento & desarrollo , Activación Enzimática , Receptores ErbB/genética , Regulación del Desarrollo de la Expresión Génica , Genes Dominantes , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Larva , Proteínas de la Membrana/metabolismo , Fenotipo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Transducción de Señal , Temperatura , Venas/crecimiento & desarrolloAsunto(s)
Proteínas de Drosophila , Inducción Embrionaria , Proteínas de Insectos/fisiología , Animales , Tipificación del Cuerpo/fisiología , Drosophila , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Metaloendopeptidasas/metabolismo , Transducción de Señal , Metaloproteinasas Similares a TolloidRESUMEN
The neighboring homologous knirps (kni) and knirps-related (knrl) genes in Drosophila encode transcription factors in the steroid hormone receptor superfamily. During early embryogenesis, kni functions as a gap gene to control expression of segmentation genes within the abdominal region of the embryo. In this study, we present evidence that kni and knrl link A/P positional information in larval wing imaginal discs to morphogenesis of the second longitudinal wing vein (L2). We show that kni and knrl are expressed in similar narrow stripes corresponding to the position of the L2 primordium. The kni and knrl L2 stripes abut the anterior border of the broad central expression domain of the Dpp target gene spalt major (salm). We provide evidence that radius incompletus (ri), a well-known viable mutant lacking the L2 vein, is a regulatory mutant of the kni/knrl locus. In ri mutant wing discs, kni and knrl fail to be expressed in the L2 primordium. In addition, the positions of molecular breakpoints in the kni/knrl locus indicate that the ri function is provided by cis-acting sequences upstream of the kni transcription unit. Epistasis tests reveal that the kni/knrl locus functions downstream of spalt major (salm) and upstream of genes required to initiate vein-versus-intervein differentiation. Mis-expression experiments suggest that kni and knrl expressing cells inhibit neighboring cells from becoming vein cells. Finally, kni and knrl are likely to refine the L2 position by positively auto-regulating their own expression and by providing negative feedback to repress salm expression. We propose a model in which the combined activities of kni and knrl organize development of the L2 vein in the appropriate position.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Proteínas de Drosophila , Drosophila/embriología , Proteínas Represoras/fisiología , Factores de Transcripción/fisiología , Alas de Animales/irrigación sanguínea , Alas de Animales/embriología , Animales , Drosophila/genética , Retroalimentación , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/fisiología , Hormonas de Insectos/fisiología , Proteínas de la Membrana/fisiología , Mutación , Secuencias Reguladoras de Ácidos NucleicosRESUMEN
Previous studies have suggested that vein primordia in Drosophila form at boundaries along the A/P axis between discrete sectors of the larval wing imaginal disc. Genes involved in initiating vein development during the third larval instar are expressed either in narrow stripes corresponding to vein primordia or in broader 'provein' domains consisting of cells competent to become veins. In addition, genes specifying the alternative intervein cell fate are expressed in complementary intervein regions. The regulatory relationships between genes expressed in narrow vein primordia, in broad provein stripes and in interveins remains unknown, however. In this manuscript, we provide additional evidence for veins forming in narrow stripes at borders of A/P sectors. These experiments further suggest that narrow vein primordia produce secondary short-range signal(s), which activate expression of provein genes in a broad pattern in neighboring cells. We also show that crossregulatory interactions among genes expressed in veins, proveins and interveins contribute to establishing the vein-versus-intervein pattern, and that control of gene expression in vein and intervein regions must be considered on a stripe-by-stripe basis. Finally, we present evidence for a second set of vein-inducing boundaries lying between veins, which we refer to as paravein boundaries. We propose that veins develop at both vein and paravein boundaries in more 'primitive' insects, which have up to twice the number of veins present in Drosophila. We present a model in which different A/P boundaries organize vein-specific genetic programs to govern the development of individual veins.
Asunto(s)
Tipificación del Cuerpo/genética , Proteínas de Drosophila , Drosophila/embriología , Venas/embriología , Alas de Animales/irrigación sanguínea , Alas de Animales/embriología , Animales , Drosophila/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas Hedgehog , Hibridación in Situ , Proteínas de Insectos/metabolismo , Modelos Biológicos , FenotipoRESUMEN
Receptor tyrosine kinase (RTK) signaling is mediated by a signaling cascade culminating in activation of mitogen-activated protein kinase (MAPK) by double phosphorylation on threonine and tyrosine residues. The pattern of MAPK activation can now be directly visualized in situ during embryonic and adult development using an antiserum is specific for the double phosphorylated form of MAPK (db-P MAPK). The pattern of MAPK activation detected by this antiserum in developing embryos and larval imaginal discs conforms remarkably well to the inferred pattern of known RTK function. In addition, db-P MAPK staining directly reveals features of signaling such as the range of signal spreading and the kinetics of RTK activation, which would be difficult to measure by other methods. The ability to visualize the output of RTK signaling also permits detailed establishment of epistatic relationships between signaling components of RTK cascades.
Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Proteínas de Drosophila , Drosophila/embriología , Embrión no Mamífero/fisiología , Proteínas Tirosina Quinasas Receptoras/metabolismo , Transducción de Señal , Animales , Receptores ErbB/metabolismo , Receptores de Factores de Crecimiento de Fibroblastos/metabolismoRESUMEN
During Drosophila embryogenesis, each segment is subdivided into an anterior and a posterior compartment through the action of the engrailed gene. Compartmental boundaries bisect imaginal disc primordia which give rise to adult appendages. In early larval development, a short-range Hedgehog signal originating from the posterior compartment of the imaginal wing disc activates expression of genes including decapentaplegic (dpp) in a stripe running along the anterior-posterior compartment boundary. Secreted Dpp emanating from the A/P boundary of wing discs then acts as a secondary signal to organize the wing over large distances. The transcription factor encoded by spalt major (salm) gene, which is expressed in a broad wedge centered over the dpp stripe, is one target of Dpp signaling. In this manuscript, we show that the anterior edge of the salm expression domain abuts a narrow stripe of rhomboid (rho)-expressing cells corresponding to the L2 longitudinal vein primordium. hh mis-expression along the anterior wing margin induces a surrounding domain of salm expression, the anterior edge of which abuts a displaced rho L2 stripe. salm plays a key role in defining the position of the L2 vein since loss of salm function in mosaic patches induces the formation of ectopic L2 branches, which comprise salm- cells running along clone borders where salm- cells confront salm+ cells. These data suggest that salm determines the position of the L2 vein primordium by activating rho expression in neighboring cells through a locally non-autonomous mechanism. rho then functions to initiate and maintain vein differentiation. We discuss how these data provide the final link connecting the formation of a linear adult structure to the establishment of a boundary by the maternal Bicoid morphogen gradient in the blastoderm embryo.
Asunto(s)
Proteínas de Drosophila , Drosophila/genética , Regulación del Desarrollo de la Expresión Génica , Genes de Insecto , Proteínas de Homeodominio/biosíntesis , Factores de Transcripción/biosíntesis , Alas de Animales/crecimiento & desarrollo , Animales , Drosophila/embriología , Drosophila/crecimiento & desarrollo , Embrión no Mamífero/fisiología , Proteínas de Insectos/biosíntesis , Mosaicismo , Factores de Transcripción/metabolismo , Factor de Crecimiento Transformador beta/biosíntesis , Alas de Animales/embriologíaRESUMEN
Contaminated water and soil at active or abandoned munitions plants is a serious problem since these compounds pose risks to human health and can be toxic to aquatic and terrestrial life. Our objective was to determine if zero-valent iron (Fe(0)) could be used to promote remediation of water and soil contaminated with 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX). As little as 1% Fe(0) (w/v) removed 70 mg TNT litre(-1) from aqueous solution within 8 h and removed 32 mg RDX litre(-1) within 96 h. Treating slurries (1:5 soil:water) of highly contaminated soil (5200 mg TNT and 6400 mg RDX kg(-1) soil) from the former Nebraska Ordnance Plant (NOP) with 10% Fe(0) (w/w soil) reduced CH(3)CN-extractable TNT and RDX concentrations below USEPA remediation goals (17.2 mg TNT and 5.8 mg RDX kg(-1)). Sequential treatment of a TNT-contaminated solution (70 mg TNT litre(-1) spiked with (14)C-TNT) with Fe(0) (5% w/v) followed by H(2)O(2) (1% v/v) completely destroyed TNT and removed about 94% of the (14)C from solution, 48% of which was mineralized to (14)CO(2) within 8 h. Fe(0)-treated TNT also was more susceptible to biological mineralization. Our observations indicate that Fe(0) alone, Fe(0) followed by H(2)O(2), or Fe(0) in combination with biotic treatment can be used for effective remediation of munitions-contaminated water and soil.
RESUMEN
TGF-beta-related signaling pathways play diverse roles during vertebrate and invertebrate development. A common mechanism for regulating the activity of TGF-beta family members is inhibition by extracellular antagonists. Recently, the Drosophila short gastrulation (sog) gene was shown to encode a predicted diffusible factor which antagonizes signaling mediated by the TGF-beta-like Decapentaplegic (Dpp) pathway in the early blastoderm embryo. sog and dpp, which are among the earliest zygotic genes to be activated, are expressed in complementary dorsal-ventral domains. The opposing actions of sog and dpp in the early embryo have been highly conserved during evolution as their vertebrate counterparts, chordin and BMP-4, function homologously to define neural versus non-neural ectoderm in Xenopus. Here we exploit the genetically sensitive adult wing vein pattern to investigate the generality of the antagonistic relationship between sog and dpp. We show that dpp is expressed in vein primordia during pupal wing development and functions to promote vein formation. In contrast, sog is expressed in complementary intervein cells and suppresses vein formation. sog and dpp function during the same phenocritical periods (i.e. 16-28 hours after pupariation) to influence the vein versus intervein cell fate choice. The conflicting activities of dpp and sog are also revealed by antagonistic dosage-sensitive interactions between these two genes during vein development. Analysis of vein and intervein marker expression in dpp and sog mutant wings suggests that dpp promotes vein fates indirectly by activating the vein gene rhomboid (rho), and that sog functions by blocking an autoactivating Dpp feedback loop. These data support the view that Sog is a dedicated Dpp antagonist.
Asunto(s)
Proteínas de Drosophila , Genes de Insecto , Proteínas de Insectos/aislamiento & purificación , Factor de Crecimiento Transformador beta/aislamiento & purificación , Alas de Animales/crecimiento & desarrollo , Animales , Tipificación del Cuerpo/genética , Drosophila/crecimiento & desarrollo , Dosificación de Gen , Proteínas de Insectos/genética , Modelos Biológicos , Mutación , Fenotipo , Pupa , Factores de Tiempo , Distribución Tisular , Factor de Crecimiento Transformador beta/genética , Alas de Animales/anatomía & histologíaRESUMEN
The short gastrulation (sog) gene is expressed in broad lateral stripes comprising the neuroectoderm of the Drosophila blastoderm embryo. sog encodes a predicted secreted protein that functions nonautonomously to antagonize the activity of the TGF-beta-like Decapentaplegic (Dpp) signaling pathway in the dorsal region of the embryo. Recently, it has been shown that sog and dpp are functionally equivalent to their respective Xenopus homologs chordin and BMP-4. In this report we provide the first direct evidence that sog plays a local role in the lateral region of the blastoderm embryo to oppose Dpp activity in the neuroectoderm. In the dorsal region, Dpp signaling both suppresses neurogenesis and maintains expression of genes that promote dorsal cell fates (dorsalization). We show that Dpp also can perform both of these functions in the neuroectoderm. In wild-type embryos, the ability of Dpp to induce expression of dorsal markers including itself (autoactivation) in the neuroectoderm is blocked by sog. We propose that Sog protects the neuroectoderm from an invasive positive feedback loop created by Dpp diffusion and autoactivation. We show that the two functions of Dpp signaling, neural suppression and dorsalization, are triggered by distinct thresholds of Dpp activity. Epistasis experiments reveal that all observed sog activity can be accounted for by Sog functioning as a dedicated Dpp antagonist. Finally, we provide evidence that Sog functions as a diffusible morphogen in the blastoderm embryo. These data strongly support the view that the primary phylogenetically conserved function of the Drosophila sog and dpp genes and the homologous Xenopus chordin and BMP-4 genes is to subdivide the primitive embryonic ectoderm into neural versus non-neural domains.
Asunto(s)
Proteínas de Drosophila , Drosophila/embriología , Genes de Insecto , Proteínas de Homeodominio , Proteínas de Insectos/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Sistema Nervioso/embriología , Animales , Blastodermo , Tipificación del Cuerpo , Proteína Morfogenética Ósea 4 , Proteínas Morfogenéticas Óseas , Comunicación Celular , Ectodermo , Epistasis Genética , Gástrula , Regulación del Desarrollo de la Expresión Génica , Glicoproteínas , Proteínas de Insectos/genética , Morfogénesis , Mutación , Proteínas Represoras , Proteínas de XenopusRESUMEN
Patterned expression of the Drosophila rhomboid (rho) gene is thought to promote signaling by the EGF receptor (EGFR) in specific cell types. In this report we examine the subcellular localization of the Rhomboid protein (Rho) which is predicted to be an integral membrane protein. At the light level, immunocytochemical staining for Rho reveals a small number of large patches (or plaques) at or near the apical cell surface. In some cells Rho plaques colocalize with Armadillo at adherens junctions, while in other cells plaques are only found basal to the adherens junction. Immunoelectron microscopy reveals that Rho plaques are composed of a highly localized patch of plasma membrane and a densely staining underlying structure. Concentration of Rho in distinct plaques depends on a balance of synthesis and membrane recycling since increasing the amount of rho expression or blocking membrane recycling leads to more uniform cell surface labeling. A limiting cellular component also appears to be required for concentrating Rho in plaques. We discuss clustering of Rho in plasma membrane patches with respect to the proposed role of Rho in promoting EGF-R signaling.
Asunto(s)
Proteínas de Drosophila , Drosophila/metabolismo , Proteínas de la Membrana/metabolismo , Animales , Membrana Celular/metabolismo , Drosophila/ultraestructura , Receptores ErbB/metabolismo , Inmunohistoquímica , Hibridación in Situ , Proteínas de la Membrana/biosíntesis , Microscopía Inmunoelectrónica , Transducción de Señal , Transcripción GenéticaRESUMEN
The Spemann organizer has long been recognized as a major source of patterning signals during the gastrula stage of amphibian embryogenesis. More recent evidence has suggested that the ventral side of the embryo also plays an important role in dorsal-ventral patterning during gastrulation through the action of signaling factors such as BMP-4. Bmp-4 is closely related to the Drosophila decapentaplegic (dpp) gene, and like Bmp-4, dpp is excluded from the neurogenic region. Recently we showed that Bmp-4 functions in an analogous role to that of dpp in Drosophila, suggesting that the mechanism of dorsal-ventral patterning in Xenopus and Drosophila embryos may be conserved. To further test this hypothesis, RNA of the Drosophila short gastrulation (sog) gene was injected into Xenopus embryos, since sog has been shown genetically to be an antagonist of dpp function. Overexpression of sog RNA in Xenopus dorsalizes the embryo by expanding neurogenic and dorsal paraxial tissue. When ectopically expressed on the ventral side of the embryo, sog induces a partial secondary axis. In addition, sog partially rescues embryos ventralized by ultraviolet irradiation. Since sog induces many similar changes in gene expression to that caused by truncated BMP receptors, we suggest that sog functions in part by opposing BMP-4 signaling. The recent identification of a possible Xenopus sog homolog, chordin, in conjunction with these results supports the hypothesis that dorsal-ventral patterning mechanisms are conserved between these two species.