RESUMEN
Phenotypic heterogeneity is very common in genetic systems and in human diseases and has important consequences for disease diagnosis and treatment. In addition to the many genetic and non-genetic (e.g., epigenetic, environmental) factors reported to account for part of the heterogeneity, we stress the importance of stochastic fluctuation and regulatory network topology in contributing to phenotypic heterogeneity. We argue that a threshold effect is a unifying principle to explain the phenomenon; that ultrasensitivity is the molecular mechanism for this threshold effect; and discuss the three conditions for phenotypic heterogeneity to occur. We suggest that threshold effects occur not only at the cellular level, but also at the organ level. We stress the importance of context-dependence and its relationship to pleiotropy and edgetic mutations. Based on this model, we provide practical strategies to study human genetic diseases. By understanding the network mechanism for ultrasensitivity and identifying the critical factor, we may manipulate the weak spot to gently nudge the system from an ultrasensitive state to a stable non-disease state. Our analysis provides a new insight into the prevention and treatment of genetic diseases.
Asunto(s)
Enfermedades Genéticas Congénitas , Mutación , Humanos , FenotipoRESUMEN
Huntington's disease (HD) belongs to protein misfolding disorders associated with polyglutamine (polyQ)-rich mutant huntingtin (mHtt) protein inclusions. Currently, it is indicated that the aggregation of polyQ-rich mHtt participates in neuronal toxicity and dysfunction. Here, we designed and synthesized a polyglutamine-specific gold nanoparticle (AuNP) complex, which specifically targeted mHtt and alleviated its toxicity. The polyglutamine-specific AuNPs were prepared by decorating the surface of AuNPs with an amphiphilic peptide (JLD1) consisting of both polyglutamine-binding sequences and negatively charged sequences. By applying the polyQ aggregation model system, we demonstrated that AuNPs-JLD1 dissociated the fibrillary aggregates from the polyQ peptide and reduced its ß-sheet content in a concentration-dependent manner. By further integrating polyethyleneimine (PEI) onto AuNPs-JLD1, we generated a complex (AuNPs-JLD1-PEI). We showed that this complex could penetrate cells, bind to cytosolic mHtt proteins, dissociate mHtt inclusions, reduce mHtt oligomers, and ameliorate mHtt-induced toxicity. AuNPs-JLD1-PEI was also able to be transported to the brain and improved the functional deterioration in the HD Drosophila larva model. Our results revealed the feasibility of combining AuNPs, JLD1s, and cell-penetrating polymers against mHtt protein aggregation and oligomerization, which hinted on the early therapeutic strategies against HD.
Asunto(s)
Materiales Biocompatibles/farmacología , Proteínas de Drosophila/antagonistas & inhibidores , Oro/farmacología , Proteína Huntingtina/antagonistas & inhibidores , Enfermedad de Huntington/tratamiento farmacológico , Nanopartículas del Metal/química , Compuestos Organometálicos/farmacología , Péptidos/farmacología , Animales , Materiales Biocompatibles/química , Drosophila , Proteínas de Drosophila/metabolismo , Oro/química , Proteína Huntingtina/metabolismo , Enfermedad de Huntington/metabolismo , Ensayo de Materiales , Compuestos Organometálicos/química , Péptidos/química , Agregado de Proteínas/efectos de los fármacosRESUMEN
The retinal basal glia (RBG) is a group of glia that migrates from the optic stalk into the third instar larval eye disc while the photoreceptor cells (PR) are differentiating. The RBGs are grouped into three major classes based on molecular and morphological characteristics: surface glia (SG), wrapping glia (WG) and carpet glia (CG). The SGs migrate and divide. The WGs are postmitotic and wraps PR axons. The CGs have giant nucleus and extensive membrane extension that each covers half of the eye disc. In this study, we used lineage tracing methods to determine the lineage relationships among these glia subtypes and the temporal profile of the lineage decisions for RBG development. We found that the CG lineage segregated from the other RBG very early in the embryonic stage. It has been proposed that the SGs migrate under the CG membrane, which prevented SGs from contacting with the PR axons lying above the CG membrane. Upon passing the front of the CG membrane, which is slightly behind the morphogenetic furrow that marks the front of PR differentiation, the migrating SG contact the nascent PR axon, which in turn release FGF to induce SGs' differentiation into WG. Interestingly, we found that SGs are equally distributed apical and basal to the CG membrane, so that the apical SGs are not prevented from contacting PR axons by CG membrane. Clonal analysis reveals that the apical and basal RBG are derived from distinct lineages determined before they enter the eye disc. Moreover, the basal SG lack the competence to respond to FGFR signaling, preventing its differentiation into WG. Our findings suggest that this novel glia-to-glia differentiation is both dependent on early lineage decision and on a yet unidentified regulatory mechanism, which can provide spatiotemporal coordination of WG differentiation with the progressive differentiation of photoreceptor neurons.
Asunto(s)
Diferenciación Celular/fisiología , Neuroglía/fisiología , Disco Óptico/embriología , Animales , Axones/metabolismo , Movimiento Celular , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Morfogénesis/fisiología , Neurogénesis/fisiología , Neuroglía/metabolismo , Neuronas/metabolismo , Disco Óptico/metabolismo , Células Fotorreceptoras de Invertebrados/fisiología , Retina/embriología , Retina/metabolismo , Transducción de Señal/fisiologíaRESUMEN
Micro-exons are exons of very small size (usually 3-30 nts). Some micro-exons are alternatively spliced. Their functions, regulation and evolution are largely unknown. Here, we present an example of an alternatively spliced 3 bp micro-exon (micro-Ex8) in the homothorax (hth) gene in Drosophila. Hth is involved in many developmental processes. It contains a MH domain and a TALE-class homeodomain (HD). It binds to another homeodomain Exd via its MH domain to promote the nuclear import of the Hth-Exd complex and serve as a cofactor for Hox proteins. The MH and HD domains in Hth as well as the HTh-Exd interaction are highly conserved in evolution. The alternatively spliced micro-exon lies between the exons encoding the MH and HD domains. We provide clear proof that the micro-Ex8 is produced by alternative splicing from a 48 bp full-length exon 8 (FL-Ex8) and the micro-Ex8 is the first three nt is FL-Ex8. We found that the micro-Ex8 is the ancient form and the 3 + 48 organization of alternatively spliced overlapping exons only emerged in the Schizophora group of Diptera and is absolutely conserved in this group. We then used several strategies to test the in vivo function of the two types of isoforms and found that the micro-Ex8 and FL-Ex8 isoforms have largely overlapping functions but also have non-redundant functions that are tissue-specific, which supports their strong evolutionary conservation. Since the different combinations of protein interaction of Hth with Exd and/or Hox can have different DNA target specificity, our finding of alternatively spliced isoforms adds to the spectrum of structural and functional diversity under developmental regulation.
Asunto(s)
Empalme Alternativo/genética , Proteínas de Drosophila/genética , Proteínas de Drosophila/fisiología , Drosophila/genética , Exones/genética , Regulación del Desarrollo de la Expresión Génica/genética , Genes de Insecto/genética , Proteínas de Homeodominio/genética , Sitios de Empalme de ARN/genética , Animales , Evolución Molecular , Proteínas de Homeodominio/fisiología , Isoformas de ProteínasRESUMEN
Photoreceptor (PR) axons project from the retina to the optic lobe in brain and form a precise retinotopic map in the Drosophila visual system. Yet the role of retinal basal glia in the retinotopic map formation is not previously known. We examined the formation of the retinotopic map by marking single PR pairs and following their axonal projections. In addition to confirming previous studies that the spatial information is preserved from the retina to the optic stalk and then to the optic lamina, we found that the young PR R3/4 axons transiently overshoot and then retract to their final destination, the lamina plexus. We then examined the process of wrapping glia (WG) membrane extension in the eye disc and showed that the WG membrane extensions also follow the retinotopic map. We show that the WG is important for the proper spatial distribution of PR axons in the optic stalk and lamina, suggesting an active role of wrapping glia in the retinotopic map formation.
Asunto(s)
Axones/ultraestructura , Drosophila melanogaster , Neuroglía , Disco Óptico/ultraestructura , Lóbulo Óptico de Animales no Mamíferos/ultraestructura , Células Fotorreceptoras de Invertebrados/ultraestructura , Animales , Drosophila melanogaster/fisiología , Drosophila melanogaster/ultraestructura , Microscopía Confocal/métodos , Microscopía Electrónica de Transmisión/métodos , Neuroglía/fisiología , Neuroglía/ultraestructura , Células Fotorreceptoras de Invertebrados/citologíaRESUMEN
Expansion of poly-glutamine (polyQ) stretches in several proteins has been linked to neurodegenerative diseases. The effects of polyQ-expanded proteins on neurons have been extensively studied, but their effects on glia remain unclear. We found that expression of distinct polyQ proteins exclusively in all glia or specifically in the blood-brain barrier (BBB) and blood-retina barrier (BRB) glia caused cell-autonomous impairment of BBB/BRB integrity, suggesting that BBB/BRB glia are most vulnerable to polyQ-expanded proteins. Furthermore, we also found that BBB/BRB leakage in Drosophila is reflected in reversed waveform polarity on the basis of electroretinography (ERG), making ERG a sensitive method to detect BBB/BRB leakage. The polyQ-expanded protein Atxn3-84Q forms aggregates, induces BBB/BRB leakage, restricts Drosophila lifespan and reduces the level of Repo (a pan-glial transcriptional factor required for glial differentiation). Expression of Repo in BBB/BRB glia can rescue BBB/BRB leakage, suggesting that the reduced expression of Repo is important for the effect of polyQ on BBB/BRB impairment. Coexpression of the chaperon HSP40 and HSP70 effectively rescues the effects of Atxn3-84Q, indicating that polyQ protein aggregation in glia is deleterious. Intriguingly, coexpression of wild-type Atxn3-27Q can also rescue BBB/BRB impairment, suggesting that normal polyQ protein may have a protective function.
Asunto(s)
Ataxina-3/genética , Enfermedades Neurodegenerativas/genética , Neuroglía/metabolismo , Péptidos/genética , Animales , Barrera Hematoencefálica/metabolismo , Barrera Hematoencefálica/patología , Barrera Hematorretinal/metabolismo , Barrera Hematorretinal/patología , Modelos Animales de Enfermedad , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Electrorretinografía , Ojo/ultraestructura , Regulación de la Expresión Génica , Proteínas del Choque Térmico HSP40/genética , Proteínas HSP70 de Choque Térmico/genética , Humanos , Enfermedades Neurodegenerativas/fisiopatología , Neuroglía/patología , Neuronas/metabolismo , Neuronas/patologíaRESUMEN
Compartment boundary formation plays an important role in development by separating adjacent developmental fields. Drosophila imaginal discs have proven valuable for studying the mechanisms of boundary formation. We studied the boundary separating the proximal A1 segment and the distal segments, defined respectively by Lim1 and Dll expression in the eye-antenna disc. Sharp segregation of the Lim1 and Dll expression domains precedes activation of Notch at the Dll/Lim1 interface. By repressing bantam miRNA and elevating the actin regulator Enable, Notch signaling then induces actomyosin-dependent apical constriction and epithelial fold. Disruption of Notch signaling or the actomyosin network reduces apical constriction and epithelial fold, so that Dll and Lim1 cells become intermingled. Our results demonstrate a new mechanism of boundary formation by actomyosin-dependent tissue folding, which provides a physical barrier to prevent mixing of cells from adjacent developmental fields.
Asunto(s)
Antenas de Artrópodos/crecimiento & desarrollo , Proteínas de Drosophila/metabolismo , Drosophila/genética , Proteínas de Homeodominio/metabolismo , Proteínas con Homeodominio LIM/metabolismo , Receptores Notch/metabolismo , Factores de Transcripción/metabolismo , Animales , Clonación Molecular , Drosophila/embriología , Proteínas de Drosophila/genética , Células Epiteliales/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Procesamiento de Imagen Asistido por Computador , Proteínas con Homeodominio LIM/genética , MicroARNs/genética , MicroARNs/metabolismo , Pliegue de Proteína , Receptores Notch/genética , Transducción de Señal , Factores de Transcripción/genética , Alas de Animales/crecimiento & desarrolloRESUMEN
Live imaging provides the ability to continuously track dynamic cellular and developmental processes in real time. Drosophila larval imaginal discs have been used to study many biological processes, such as cell proliferation, differentiation, growth, migration, apoptosis, competition, cell-cell signaling, and compartmental boundary formation. However, methods for the long-term ex vivo culture and live imaging of the imaginal discs have not been satisfactory, despite many efforts. Recently, we developed a method for the long-term ex vivo culture and live imaging of imaginal discs for up to 18 h. In addition to using a high insulin concentration in the culture medium, a low-melting agarose was also used to embed the disc to prevent it from drifting during the imaging period. This report uses the eye-antennal discs as an example. Photoreceptor R3/4-specific mδ0.5-Ga4 expression was followed to demonstrate that photoreceptor differentiation and ommatidial rotation can be observed during a 10 h live imaging period. This is a detailed protocol describing this simple method.
Asunto(s)
Ojo/embriología , Discos Imaginales/citología , Animales , Diferenciación Celular , Medios de Cultivo , Drosophila , Proteínas de Drosophila/metabolismo , Ojo/metabolismo , Discos Imaginales/embriología , Insulina/farmacología , Larva/citología , Larva/metabolismo , SefarosaRESUMEN
5D images of engrailed (en) and eye gone (eyg) gene expressions during the course of the eye-antenna disc primordium (EADP) formation of Drosophila embryos from embryonic stages 13 through 16 were recorded via light sheet microscopy and analyzed to reveal the cell dynamics involved in the development of the EADP. Detailed analysis of the time-lapsed images revealed the process of EADP formation and its invagination trajectory, which involved an inversion of the EADP anterior-posterior axis relative to the body. Furthermore, analysis of the en-expression pattern in the EADP provided strong evidence that the EADP is derived from one of the en-expressing head segments.
Asunto(s)
Drosophila melanogaster/genética , Ojo/metabolismo , Microscopía Fluorescente/métodos , Animales , Biomarcadores , Drosophila melanogaster/metabolismo , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Genes Reporteros , Imagenología TridimensionalRESUMEN
Neurons and glia interact reciprocally. Glia perform many important functions in the development and proper functioning of the nervous system throughout different stages of life. Neurons also affect the development and function of glia. Neurodegenerative diseases are usually late-onset, progressive, and affect specific parts of the nervous system. Many neurodegenerative disorders have been extensively studied. However, the majority of the studies have focused on the events that occur in neurons. The events that occur in glia, and whether and how glia participate in the pathogenesis of these diseases, have not been as well studied. In this review, we will focus on how the fruit fly Drosophila melanogaster has been used as a model to study neuron-glia interactions in neurodegenerative disorders. We discuss how glia are affected in these models of human neurodegenerative disorders and how glia contribute to their pathogenesis. These studies have provided important insight into the mechanisms of diverse neurodegenerative disorders, and have suggested possibilities for early diagnosis.
Asunto(s)
Degeneración Nerviosa/patología , Enfermedades Neurodegenerativas/patología , Neuroglía/patología , Neuronas/patología , Animales , Modelos Animales de Enfermedad , DrosophilaRESUMEN
The late onset of neurodegeneration in humans indicates that the survival and function of cells in the nervous system must be maintained throughout adulthood. In the optic lamina of the adult Drosophila, the photoreceptor axons are surrounded by multiple types of glia. We demonstrated that the adult photoreceptors actively contribute to glia maintenance in their target field within the optic lamina. This effect is dependent on the epidermal growth factor receptor (EGFR) ligands produced by the R1-6 photoreceptors and transported to the optic lamina to act on EGFR in the lamina glia. EGFR signaling is necessary and sufficient to act in a cell-autonomous manner in the lamina glia. Our results suggest that EGFR signaling is required for the trafficking of the autophagosome/endosome to the lysosome. The loss of EGFR signaling results in cell degeneration most likely because of the accumulation of autophagosomes. Our findings provide in vivo evidence for the role of adult neurons in the maintenance of glia and a novel role for EGFR signaling in the autophagic flux.
Asunto(s)
Diferenciación Celular/genética , Receptores ErbB/genética , Neuroglía/metabolismo , Retina/metabolismo , Animales , Axones/metabolismo , Drosophila/crecimiento & desarrollo , Drosophila/metabolismo , Humanos , Neuronas/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Retina/patología , Transducción de Señal , Asta Dorsal de la Médula Espinal/crecimiento & desarrollo , Asta Dorsal de la Médula Espinal/metabolismoRESUMEN
Organ formation requires a delicate balance of positive and negative regulators. In Drosophila eye development, wingless (wg) is expressed at the lateral margins of the eye disc and serves to block retinal development. The T-box gene optomotor-blind (omb) is expressed in a similar pattern and is regulated by Wg. Omb mediates part of Wg activity in blocking eye development. Omb exerts its function primarily by blocking cell proliferation. These effects occur predominantly in the ventral margin. Our results suggest that the primary effect of Omb is the blocking of Jak/STAT signaling by repressing transcription of upd which encodes the Jak receptor ligand Unpaired.
Asunto(s)
Proliferación Celular/fisiología , Proteínas de Drosophila/metabolismo , Ojo/embriología , Quinasas Janus/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Factores de Transcripción STAT/metabolismo , Proteínas de Dominio T Box/metabolismo , Factores de Transcripción/metabolismo , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster , Ojo/citología , Quinasas Janus/genética , Proteínas del Tejido Nervioso/genética , Factores de Transcripción STAT/genética , Proteínas de Dominio T Box/genética , Factores de Transcripción/genéticaRESUMEN
A general question in development is how do adjacent primordia adopt different developmental fates and stably maintain their distinct fates? In Drosophila melanogaster, the adult eye and antenna originate from the embryonic eye-antenna primordium. These cells proliferate in the larval stage to form the eye-antenna disc. The eye or antenna differs at mid second instar with the restricted expression of Cut (Ct), a homeodomain transcriptional repressor, in the antenna disc and Eyeless (Ey), a Pax6 transcriptional activator, in the eye disc. In this study, we show that ey transcription in the antenna disc is repressed by two homeodomain proteins, Ct and Homothorax (Hth). Loss of Ct and Hth in the antenna disc resulted in ectopic eye development in the antenna. Conversely, the Ct and Hth expression in the eye disc was suppressed by the homeodomain transcription factor Sine oculis (So), a direct target of Ey. Loss of So in the eye disc caused ectopic antenna development in the eye. Therefore, the segregation of eye and antenna fates is stably maintained by mutual repression of the other pathway.
Asunto(s)
Antenas de Artrópodos/embriología , Antenas de Artrópodos/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Ojo/embriología , Ojo/metabolismo , Animales , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The Drosophila head vertex is composed of three ocelli, stereotypic bristle patterns and characteristic cuticles. It is derived from the fusion of two eye-antenna discs. The head vertex primordium is located at the anterior-dorsal region of the eye disc. The orthodenticle (otd) homeobox gene is expressed in the primordium and is functionally required for its development and patterning. Here we show that the Pax gene eye gone (eyg) is expressed adjacent to the otd expression domain in the eye disc. otd is required and sufficient to repress eyg transcription, thereby preventing eyg from expressing in the head vertex primordium. In otd mutant, eyg expression is derepressed in the head vertex primordium and is a major negative effector to block head vertex development. Therefore, otd not only needs to induce downstream effector genes to execute the development and patterning of the head vertex development, but also needs to actively repress the negative regulator eyg. In addition, eyg is required for the development of the lateral bristles in the head vertex. So eyg plays both positive and negative roles in head vertex development.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Drosophila melanogaster/genética , Genes de Insecto/genética , Animales , Drosophila melanogaster/citología , Elementos de Facilitación Genéticos/genética , Regulación del Desarrollo de la Expresión Génica , Cabeza/embriología , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Modelos Genéticos , Especificidad de Órganos/genética , Fenotipo , Transducción de Señal/genética , Transcripción Genética , Proteína Wnt1/genética , Proteína Wnt1/metabolismoRESUMEN
We show here that the Drosophila MADF/BESS domain transcription factor Dip3, which is expressed in differentiating photoreceptors, regulates neuronal differentiation in the compound eye. Loss of Dip3 activity in photoreceptors leads to an extra photoreceptor in many ommatidia, while ectopic expression of Dip3 in non-neuronal cells results in photoreceptor loss. These findings are consistent with the idea that Dip3 is required non-cell autonomously to block extra photoreceptor formation. Dip3 may mediate the spatially restricted potentiation of Notch (N) signaling since the Dip3 misexpression phenotype is suppressed by reducing N signaling and misexpression of Dip3 leads to ectopic activity of a N-responsive enhancer. Analysis of mosaic ommatidia suggests that no specific photoreceptor must be mutant to generate the mutant phenotype. Remarkably, however, mosaic pupal ommatidia with three or fewer Dip3(+) photoreceptors always differentiate an extra photoreceptor, while those with four or more Dip3(+) photoreceptors never differentiate an extra photoreceptor. These findings are consistent with the notion that Dip3 in photoreceptors activates a heretofore unsuspected diffusible ligand that may work in conjunction with the N pathway to prevent a subpopulation of undifferentiated cells from choosing a neuronal fate.
Asunto(s)
Proteínas de Drosophila/fisiología , Regulación del Desarrollo de la Expresión Génica , Células Fotorreceptoras de Invertebrados/crecimiento & desarrollo , Factores de Transcripción/fisiología , Animales , Drosophila/embriología , Drosophila/genética , Drosophila/crecimiento & desarrollo , Drosophila/fisiología , Proteínas de Drosophila/genética , Embrión no Mamífero , Proteínas Fluorescentes Verdes/metabolismo , Inmunohistoquímica , Modelos Biológicos , Mutación , Neuronas/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Células Fotorreceptoras de Invertebrados/fisiología , Factores de Transcripción/genéticaRESUMEN
The Pax gene eyg is important for Drosophila eye development. eyg expression in the visual system changes dynamically during development. In this study, we found that the transcriptional regulation of eyg can be separated into four distinct temporal phases (E, L1, L2, and L3) and each is regulated by distinct cis-regulatory elements. Utilizing these enhancers for temporal and spatially specific manipulations, we addressed the regulation and function of eyg at different developmental stages. We found that Notch signaling is required and sufficient for eyg expression and this activity is restricted only to the L2 stage. We further showed that the function of eyg in eye development is required only at the second instar larval stage, while its function for head and antenna development can be provided at any time during embryo and larval development. Thus there is a temporal switch of the regulatory mechanism and function of eyg. We propose that eyg expression at L2 is induced and maintained by N signaling, and is turned off at L3 by a negative feedback loop involving the morphogenetic furrow.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila/embriología , Ojo/embriología , Regulación del Desarrollo de la Expresión Génica/fisiología , Elementos Reguladores de la Transcripción/fisiología , Animales , Inmunohistoquímica , Hibridación in Situ , Receptores Notch/metabolismo , Transducción de Señal/fisiologíaRESUMEN
In eye development the tasks of tissue specification and cell proliferation are regulated, in part, by the Pax6 and Pax6(5a) proteins respectively. In vertebrates, Pax6(5a) is generated as an alternately spliced isoform of Pax6. This stands in contrast to the fruit fly, Drosophila melanogaster, which has two Pax6(5a) homologs that are encoded by the eyegone and twin of eyegone genes. In this report we set out to determine the respective contributions that each gene makes to the development of the fly retina. Here we demonstrate that both eyg and toe encode transcriptional repressors, are expressed in identical patterns but at significantly different levels. We further show, through a molecular dissection of both proteins, that Eyg makes differential use of several domains when compared to Toe and that the number of repressor domains also differs between the two Pax6(5a) homologs. We predict that these results will have implications for elucidating the functional differences between closely related members of other Pax subclasses.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/crecimiento & desarrollo , Drosophila melanogaster/genética , Proteínas del Ojo/genética , Ojo/crecimiento & desarrollo , Genes de Insecto , Proteínas de Homeodominio/genética , Factor de Transcripción PAX5/genética , Factores de Transcripción Paired Box/genética , Proteínas Represoras/genética , Animales , Animales Modificados Genéticamente , Secuencia de Bases , ADN/genética , ADN/metabolismo , Cartilla de ADN/genética , Proteínas de Unión al ADN/química , Proteínas de Drosophila/química , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Ojo/embriología , Ojo/metabolismo , Proteínas del Ojo/química , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/química , Hibridación in Situ , Factor de Transcripción PAX5/química , Factor de Transcripción PAX6 , Factores de Transcripción Paired Box/química , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Represoras/químicaRESUMEN
In Drosophila, the eye and antenna originate from a single epithelium termed the eye-antennal imaginal disc. Illumination of the mechanisms that subdivide this epithelium into eye and antenna would enhance our understanding of the mechanisms that restrict stem cell fate. We show here that Dip3, a transcription factor required for eye development, alters fate determination when misexpressed in the early eye-antennal disc, and have taken advantage of this observation to gain new insight into the mechanisms controlling the eye-antennal switch. Dip3 misexpression yields extra antennae by two distinct mechanisms: the splitting of the antennal field into multiple antennal domains (antennal duplication), and the transformation of the eye disc to an antennal fate. Antennal duplication requires Dip3-induced under proliferation of the eye disc and concurrent over proliferation of the antennal disc. While previous studies have shown that overgrowth of the antennal disc can lead to antennal duplication, our results show that overgrowth is not sufficient for antennal duplication, which may require additional signals perhaps from the eye disc. Eye-to-antennal transformation appears to result from the combination of antennal selector gene activation, eye determination gene repression, and cell cycle perturbation in the eye disc. Both antennal duplication and eye-to-antennal transformation are suppressed by the expression of genes that drive the cell cycle providing support for tight coupling of cell fate determination and cell cycle control. The finding that this transformation occurs only in the eye disc, and not in other imaginal discs, suggests a close developmental and therefore evolutionary relationship between eyes and antennae.
Asunto(s)
Drosophila melanogaster/embriología , Ojo/embriología , Expresión Génica , Órganos de los Sentidos/embriología , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Inmunohistoquímica , Factores de Transcripción/genéticaRESUMEN
The initiation of retinal development in Drosophila begins at the posterior center (PC) of the eye disc margin. The front of the differentiation wave, recognized as a morphogenetic furrow (MF), moves from posterior to anterior. What determines MF initiates from the specific PC site is still unclear. The unpaired (upd) gene is expressed at PC at early third instar, just before the time of MF initiation. Therefore, upd is expressed at the appropriate time and location for a specific role in defining the site of MF initiation. upd encodes a ligand for the Jak/STAT signaling pathway. In this report, we showed that the Upd/Jak/STAT signaling is required and sufficient to determine MF initiation. This is primarily achieved by repressing the transcription of wingless (wg), which is known to block MF initiation.