Percorrer por autor "Silva, Nadia"
A mostrar 1 - 10 de 16
Resultados por página
Opções de ordenação
- Cellular morphology and markers of cartilage and bone in the marine teleost Sparus auratusPublication . Estêvão, M. Dulce; Silva, Nadia; Redruello, Begoña; Costa, Rita; Gregorio, Silvia; Canario, Adelino V. M.; Power, DeborahModifications have been characterised in terms of cellular organisation and the extracellular matrix (ECM) during bone ontogeny in the sea bream (Sparus auratus). During endochondral development, the agglomeration of matrix-secreting cells gives rise to chondrones; these chondrones frequently contain proliferating-cell-nuclearantigen-positive cells, which subsequently become large collagen-II-positive cells with the characteristics of chondrocytes. Moreover, the matrix:cell ratio within the perichondrium increases, accompanied by a modification in ECM composition. Mineralisation of cartilage ECM is marked by a rapid fall in cell number, the switching off of collagen II transcription and the switching on of collagen X transcription, followed by collagen I transcription and bone mineralisation. The formation of dermal structures initiated upon the condensation of mesenchyme cells defines the future location of the dermal bone. Subsequent cellular differentiation gives rise to cells on the bone surface; these cells are positive for collagen I and osteonectin transcripts. The fish skeleton, with the exception of vertebrae, tends to comprise flattened bones that are covered by a monolayer of cells, the periosteum. A third type of tissue, present in gills, consists of chondrocyte-like cells embedded in a mineralised matrix resembling chondroid bone in mammals. The results suggest that the cellular organisation and ontogeny of endochondral and dermal bone in the sea bream are similar to those described in other vertebrates.
- Corrigendum: in a zebrafish biomedical model of human Allan-Herndon-Dudley syndrome impaired MTH signaling leads to decreased neural cell diversityPublication . Silva, Nadia; Campinho, Marco António
- Differentiation of vascular characteristics using contrast-enhanced ultrasound imagingPublication . Butler, Mairead; Perperidis, Antonios; Zahra, Jean-Luc Matteo; Silva, Nadia; Averkiou, Michalakis; Duncan, W. Colin; McNeilly, Alan; Sboros, VassilisUltrasound contrast imaging has been used to assess tumour growth and regression by assessing the flow through the macro- and micro-vasculature. Our aim was to differentiate the blood kinetics of vessels such as veins, arteries and microvasculature within the limits of the spatial resolution of contrast-enhanced ultrasound imaging. The highly vascularised ovine ovary was used as a biological model. Perfusion of the ovary with SonoVue was recorded with a Philips iU22 scanner in contrast imaging mode. One ewe was treated with prostaglandin to induce vascular regression. Time-intensity curves (TIC) for different regions of interest were obtained, a lognormal model was fitted and flow parameters calculated. Parametric maps of the whole imaging plane were generated for 2 × 2 pixel regions of interest. Further analysis of TICs from selected locations helped specify parameters associated with differentiation into four categories of vessels (arteries, veins, medium-sized vessels and micro-vessels). Time-dependent parameters were associated with large veins, whereas intensity-dependent parameters were associated with large arteries. Further development may enable automation of the technique as an efficient way of monitoring vessel distributions in a clinical setting using currently available scanners.
- Effects of dietary phosphorus and calcium level on growth and skeletal development in rainbow trout (Oncorhynchus mykiss) fryPublication . Fontagné, Stéphanie; Silva, Nadia; Bazin, Didier; Ramos, Angela; Aguirre, Peyo; Surget, Anne; Abrantes, António; Kaushik, Sadasivam J.; Power, DeborahThe effects of dietary levels of phosphorus (P) and calcium(Ca) on skeletal development and mineral deposition in rainbow trout(Oncorhynchus mykiss) fry were studied. Six semi-purified diets were formulated with graded levels of P and Ca. The basal diet A contained only P supplied by casein at 0.5% of drymatter. Other diets B, C,D and Ewere supplementedwith 0.4, 0.8, 1.2 and 1.6% P supplied as a 1:1 mixture of NaH2PO4/KH2PO4 resulting in 0.8, 1.2, 1.6 and 2.2% total P, respectively. These five dietswere supplementedwith 1% Ca supplied as CaCO3whereas another diet F, supplementedwith 0.8% P,was Ca-free. Each dietwas distributed to 3 replicate tanks of 600 swimup fry (initialmean weight: 0.1 g) at a water temperature of 17 °C over a 12-week growth trial. Fish were handfed 6 times a day to visual satiety.
- Effects of pristine or contaminated polyethylene microplastics on zebrafish developmentPublication . Tarasco, Marco; Gavaia, Paulo; Bensimon-Brito, Anabela; Cordelières, Fabrice P.; Santos, Tamara; Martins, Gil; De Castro, Daniela; Silva, Nadia; Cabrita, Elsa; Bebianno, Maria; Stainier, Didier Y.R.; Cancela, M. Leonor; Laizé, VincentThe presence of microplastics in the aquatic ecosystem represents a major issue for the environment and human health. The capacity of organic pollutants to adsorb onto microplastic particles raises additional concerns, as it creates a new route for toxic compounds to enter the food web. Current knowledge on the impact of pristine and/or contaminated microplastics on aquatic organisms remains insufficient, and we provide here new insights by evaluating their biological effects in zebrafish (Danio rerio). Zebrafish larvae were raised in ZEB316 stand-alone housing systems and chronically exposed throughout their development to polyethylene particles of 20-27 mu m, pristine (MP) or spiked with benzo[alpha]pyrene (MP-BaP), supplemented at 1% w/w in the fish diet. While they had no effect at 30 days post-fertilization (dpf), MP and MP-BaP affected growth parameters at 90 and 360 dpf. Relative fecundity, egg morphology, and yolk area were also impaired in zebrafish fed MP-BaP. Zebrafish exposed to experimental diets exhibited an increased incidence of skeletal deformities at 30 dpf as well as an impaired development of caudal fin/scales, and a decreased bone quality at 90 dpf. An intergenerational bone formation impairment was also observed in the offspring of parents exposed to MP or MP-BaP through a reduction of the opercular bone in 6 dpf larvae. Beside a clear effect on bone development, histological analysis of the gut revealed a reduced number of goblet cells in zebrafish fed MP-BaP diet, a sign of intestinal inflam-mation. Finally, exposure of larvae to MP-BaP up-regulated the expression of genes associated with the BaP response pathway, while negatively impacting the expression of genes involved in oxidative stress. Altogether, these data suggest that long-term exposure to pristine/contaminated microplastics not only jeopardizes fish growth, reproduction performance, and skeletal health, but also causes intergenerational effects.
- Expression of the myosin light chains 1 and 2 in the developing fast muscle of gilthead sea bream (Sparus aurata)Publication . Moutou, Katerina A.; Silva, Nadia; Mamuris, Z.; Power, DeborahMyosin, the major component of striated muscle, is a complex molecule of heavy and light chains, which undergo continuous replacement to meet developmental and environmental demands. A range of myosin isoforms are expressed in early developmental stages and are of special interest as they offer information about muscle formation and function early in life. In addition, they can act as markers for the study of prenatal events with an effect on postnatal growth performance. In this study, the spatial and temporal expression of embryonic myosin light chains 1 (MLC1) and 2 (MLC2) was studied in sea bream larvae post-hatch by in situ hybridization using riboprobes.
- Expression patterns of MLC isoforms during halibut (Hippoglossus hippoglossus L.) metamorphosisPublication . Silva, Nadia; Baptista, Vitor M.; Power, DeborahAtlantic halibut is an important commercial fish in the countries of the North Atlantic and is emerging as a promising species for marine cold-water aquaculture. The axial musculature of the developing larvae is the largest and most rapidly growing tissue and during the transition from larval to adult muscle fibre types significant changes in fibre morphology and gene transcription occur. In fact the change in myotome height correlates well with different larval halibut stages. In the present study the spatial and temporal expression of myosin light chain 1 (MLC1), 2 (MLC2) and 3 (MLC3) was studied in metamorphosing halibut by in situ hybridization. As a first step to establishing a role for the thyroid axis in halibut muscle development whole body thyroid hormone (TH) concentrations were also determined. In first feeding larvae MLC1, MLC2A and MLC3 transcripts had a similar distribution and were confined to the muscle fibres of the germinal zones. In pre-metamorphic larvae transcripts were highly expressed throughout the epaxial and hypaxial musculature and expression levels reached a maximum in larvae starting metamorphosis, this change coincided with a significant increase in the concentration of thyroid hormones. By the time larvae reached the metamorphic climax, MLC1, MLC2A and MLC3 expression was still high throughout the musculature but expression was confined to fibres adjacent to the myosepts and to small cells scattered in the musculature, possibly satellite cells. MLC2A was also expressed in the red muscle fibres; no transition between larval and adult MLC isoforms was detected.
- Four stanniocalcin genes in teleost fish: Structure, phylogenetic analysis, tissue distribution and expression during hypercalcemic challengePublication . Schein, V; Schein, Vanessa; Cardoso, João CR; Pinto, Patricia IS; Anjos, Liliana; Silva, Nadia; Power, Deborah; Canario, Adelino V. M.Stanniocalcin (STC), first isolated from the corpuscles of Stannius (CS) of teleost fishes and a systemic regulator of mineral metabolism, is present in all vertebrates as two isoforms, STC1 and STC2, encoded by separate genes. Here we show that the genome of Tetraodon nigroviridis, and other teleosts, possess duplicate genes for each STC isoform, designated stc1-a and -b, and stc2-a and -b. Stc1-a was cloned from CS, stc2-a from muscle and the two novel cDNAs, stc1-b and stc2-b, from brain. However, stc2-b was isolated as a conjoined (read-through) transcript with bod1 (bi-orientation defective 1, or FAM44B), and two additional alternative conjoined transcripts were also isolated. The predicted STC products shared the typical vertebrate 10 conserved cysteine residues and N-linked glycosylation motifs, in addition to specific features. Gene structure was generally conserved with four exons and three introns with the exception of stc1-a which gained an extra intron in exon three, originating one extra exon. Gene order and synteny is also maintained across vertebrates and the cpeb4 gene identified in the homologue region of the chordate Ciona was linked to vertebrate stc2 but not stc1. Immunohistochemistry in different species revealed that STC1-A was found only in CS and in a few cells in kidney. STC1-B had a restricted expression and was more prominent in the gills. STC2-A was detected in a variety of tissues, including pituitary, with most abundant immunoreaction in kidney cells and gill rakers and the CS was negative. Expression of stc1-a in CS of Tetraodon was 15-fold (p < 0.05) up-regulated 2 h after transfer from 2.9 mM Ca2+ to 10 mM Ca2+ water and down-regulated after 12 hours to 11-fold lower than 2.9 mM Ca2+ fish (p < 0.05). With the exception of stc1-a in CS, low expression levels and high individual variation were generally found for the expression of stc transcripts in kidney and gills, with no statistically significant changes in response to the hypercalcemic shock. In conclusion, both stc1 and stc2 genes are represented by paralogues in teleosts genomes and the analysis performed suggests that only stc1-a in the CS is involved in extracellular calcium regulation. The widespread distribution of stcs in fish tissues supports pleiotropic roles.
- In a zebrafish biomedical model of human Allan-Herndon-Dudley syndrome impaired MTH signaling leads to decreased neural cell diversityPublication . Silva, Nadia; Campinho, Marco AntónioMaternally derived thyroid hormone (T3) is a fundamental factor for vertebrate neurodevelopment. In humans, mutations on the thyroid hormones (TH) exclusive transporter monocarboxylic acid transporter 8 (MCT8) lead to the Allan-Herndon-Dudley syndrome (AHDS). Patients with AHDS present severe underdevelopment of the central nervous system, with profound cognitive and locomotor consequences. Functional impairment of zebrafish T3 exclusive membrane transporter Mct8 phenocopies many symptoms observed in patients with AHDS, thus providing an outstanding animal model to study this human condition. In addition, it was previously shown in the zebrafish mct8 KD model that maternal T3 (MTH) acts as an integrator of different key developmental pathways during zebrafish development. MethodsUsing a zebrafish Mct8 knockdown model, with consequent inhibition of maternal thyroid hormones (MTH) uptake to the target cells, we analyzed genes modulated by MTH by qPCR in a temporal series from the start of segmentation through hatching. Survival (TUNEL) and proliferation (PH3) of neural progenitor cells (dla, her2) were determined, and the cellular distribution of neural MTH-target genes in the spinal cord during development was characterized. In addition, in-vivo live imaging was performed to access NOTCH overexpression action on cell division in this AHDS model. We determined the developmental time window when MTH is required for appropriate CNS development in the zebrafish; MTH is not involved in neuroectoderm specification but is fundamental in the early stages of neurogenesis by promoting the maintenance of specific neural progenitor populations. MTH signaling is required for developing different neural cell types and maintaining spinal cord cytoarchitecture, and modulation of NOTCH signaling in a non-autonomous cell manner is involved in this process. DiscussionThe findings show that MTH allows the enrichment of neural progenitor pools, regulating the cell diversity output observed by the end of embryogenesis and that Mct8 impairment restricts CNS development. This work contributes to the understanding of the cellular mechanisms underlying human AHDS.
- Molecular, cellular and histological changes in skin from a larval to an adult phenotype during bony fish metamorphosisPublication . Campinho, Marco António; Silva, Nadia; Sweeney, Glen E.; Power, DeborahDevelopmental models for skin exist in terrestrial and amphibious vertebrates but there is a lack of information in aquatic vertebrates. We have analysed skin epidermal development of a bony fish (teleost), the most successful group of extant vertebrates. A specific epidermal type I keratin cDNA (hhKer1), which may be a bony-fishspecific adaptation associated with the divergence of skin development (scale formation) compared with other vertebrates, has been cloned and characterized. The expression of hhKer1 and collagen 1α1 in skin taken together with the presence or absence of keratin bundle-like structures have made it possible to distinguish between larval and adult epidermal cells during skin development. The use of a flatfish with a well-defined larval to juvenile transition as a model of skin development has revealed that epidermal larval basal cells differentiate directly to epidermal adult basal cells at the climax of metamorphosis. Moreover,hhKer1 expression is downregulated at the climax of metamorphosis and is inversely correlated with increasing thyroxin levels. We suggest that, whereas early mechanisms of skin development between aquatic and terrestrial vertebrates are conserved, later mechanisms diverge.