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Sanchez-Herrero, E.

Publications and source records attributed to Sanchez-Herrero, E..

6 recordsLinked to original sources

Drosophila male genitalia rotation depends on permissive remodeling of the posterior abdomen

One of the most characteristic morphogenetic processes in Drosophila is the 360{degrees} rotation of the male pupal genital disc. This movement is driven by the myosin Myo1D, whose expression in the genital disc is controlled by the Hox gene Abdominal-B. The rotation takes place in contact and relative to the posterior abdomen, yet the contribution of abdominal tissues has remained unclear. Here we show that normal genital disc circumrotation requires active remodeling of posterior abdominal larval epidermal cells that contact the rotating terminalia. Preventing apoptosis in these cells, or increasing EGFR signaling, delays their extrusion and results in incomplete rotation without altering rotational chirality. In parallel, elimination of Extracellular Matrix by Metalloproteinase 1 in these cells, although without leading to their extrusion, is also strictly required for genital disc circumrotation. Inhibition of this metalloproteinase activity leads to persistence of collagen IV and incomplete rotation, revealing an independent requirement for Extracellular Matrix clearance at the disc-abdomen interface. By contrast, genetic conditions that prevent formation or elimination of the male A7 segment do not necessarily impair genital disc rotation, demonstrating that A7 suppression and circumrotation are separable processes. These findings identify posterior abdominal tissue remodeling as an essential extrinsic requirement that enables genital disc circumrotation.

developmental biology↗

Regeneration can take place across Drosophila compartments or segments with different Hox gene expression

Regeneration in Drosophila has primarily been studied in imaginal discs, which are organized into distinct compartments defined by strict lineage boundaries. While cells typically do not cross these borders, regeneration can bridge them to restore the original organ specification, which is determined by Hox genes. Although differences in Hox gene expression are known to segregate cell populations, the role of such differences as a potential barrier to regeneration remains unclear. We investigated this by analyzing two experimental settings: the analia (derived from the Hox-compound genital disc) and haltere discs with mutations in Ultrabithorax regulatory regions (bithorax or postbithorax). Our findings demonstrate that Hox gene differences are not an absolute impediment to regeneration across segments or compartments. However, we observed occasional regenerative limits, which were non-specifically enhanced in the postbithorax background.

developmental biology↗

The Homeodomain-interacting protein kinase Hipk promotes apoptosis by stabilizing the active form of Dronc

Members of the evolutionarily conserved homeodomain-interacting protein kinase (Hipk) family play a critical role in regulating essential signalling pathways involved in growth, differentiation, and apoptosis. While vertebrates have multiple hipk genes, Drosophila contains a single hipk ortholog, what facilitates functional analysis. We find that hipk is necessary for the stabilization of the initiator caspase Dronc, thus enhancing the two Dronc activities in apoptotic scenarios: the induction of the caspase cascade, and the reinforcement of JNK signalling pathway. Conversely, our data suggest that Dronc also raises the expression levels of Hipk, thereby reinforcing the apoptotic response. These findings significantly enhance our understanding of caspase regulation and position Hipk as a promising target for modulating caspase activity in a variety of biological contexts.

developmental biology↗

Mechanical coupling between dorsal and ventral surfaces shapes the Drosophila haltere

The extracellular matrix is an essential determinant of animal form, enabling organization of cells and tissues into organs with complex 3D shapes. In contrast with the dorso-ventrally flat Drosophila wing, its serial homolog the haltere adopts a globular shape thought to arise from a lack of matrix-mediated adhesion between its dorsal and ventral surfaces. Contradicting this model, however, matrix manipulations are known to deform halteres. To understand haltere morphogenesis, we characterized matrix behavior and monitored metamorphic development of the haltere. We found that, similar to the wing, correct haltere morphogenesis requires Collagen IV degradation, which we show is mediated by ecdysone-controlled expression of Matrix metalloprotease 2 in both wing and haltere. After Collagen IV is degraded, similar again to the wing, dorsal and ventral haltere surfaces establish Laminin-mediated contact through long cytoskeletal projections. Furthermore, time-lapse analysis of shape changes in wild type and mutant halteres indicates that these projections couple the two surfaces through a central tensor, ensuring load distribution across the whole organ to create a globular shape against tissue-wide deforming forces. Our findings reveal an unexpected role for matrix-mediated adhesion in haltere morphogenesis and describe a novel type of matrix-based tensor structure building 3D shape from 2D epithelia.

developmental biology↗

Ambivalent partnership of the Drosophila posterior class Hox protein Abdominal-B with the Extradenticle and Homothorax cofactors

Hox proteins, a sub-group of the homeodomain (HD) transcription factor family, provide positional information for axial patterning in development and evolution. Hox protein functional specificity is reached, at least in part, through Pbc (Extradenticle (Exd) in Drosophila) and Meis/Prep (Homothorax (Hth) in Drosophila) cofactor interactions. Most of our current knowledge of Hox protein specificity stems from the study of anterior and central Hox proteins, identifying the molecular and structural bases for Hox/Pbc/Meis-Prep cooperative action. Posterior Hox class proteins, Abd-B in Drosophila and Hox9-13 in vertebrates, have been comparatively less studied. They strongly diverge from anterior and central class Hox proteins, with a low degree of HD sequence conservation and the absence of a core canonical Pbc interaction motif. Here we explore how Abd-B function interface with that of Exd/Hth using several developmental contexts, studying mutual expression control, functional dependency and intrinsic protein requirements. Results identify cross regulatory interactions setting relative expression and activity levels required for proper development. They also reveal organ-specific requirement and a binary functional interplay with Exd and Hth, either synergistic or antagonistic. This highlights context specific use of Exd/Hth cofactors, and a similar context specific use of Abd-B protein intrinsic protein requirements.

developmental biology↗

A simple immunoassay for extracellular vesicle liquid biopsy in microliters of non-processed plasma

Extracellular vesicles (EVs), released by most cell types, provide an excellent source of biomarkers in biological fluids. Here we describe a method that, using just a few microliters of patients plasma, identifies tumour markers exposed on EVs. Studying physico-chemical properties of EVs in solution, we demonstrate that they behave as stable colloidal suspensions and therefore, in immunocapture assays, many of them are unable to interact with a stationary functionalised surface. Using flocculation methods, like those used to destabilize colloids, we demonstrate that cationic polymers increase EV {zeta}-potential, diameter, and sedimentation coefficient and thus, allow a more efficient capture on antibody-coated surfaces by both ELISA and bead-assisted flow cytometry. These findings led to optimization of a protocol in microtiter plates allowing effective immunocapture of EVs, directly in plasma without previous ultracentrifugation or other EV enrichment. The method, easily adaptable to any laboratory, has been validated using plasma from lung cancer patients in which the epithelial cell marker EpCAM has been detected on EVs. This high throughput, easy to automate, technology allows screening of large numbers of patients to phenotype tumour markers in circulating EVs, breaking barriers for the validation of proposed EV biomarkers and the discovery of new ones.

biochemistry↗