Loading...
15 results
Search Results
Now showing 1 - 10 of 15
- Vertebrate SLRP family evolution and the subfunctionalization of osteoglycin gene duplicates in teleost fishPublication . Costa, Rita; Brazona, Rute Sofia Tavares Martins; Capilla, E.; Anjos, Liliana; Power, DeborahBackground Osteoglycin (OGN, a.k.a. mimecan) belongs to cluster III of the small leucine-rich proteoglycans (SLRP) of the extracellular matrix (ECM). In vertebrates OGN is a characteristic ECM protein of bone. In the present study we explore the evolution of SLRP III and OGN in teleosts that have a skeleton adapted to an aquatic environment. Results The SLRP gene family has been conserved since the separation of chondrichthyes and osteichthyes. Few gene duplicates of the SLRP III family exist even in the teleosts that experienced a specific whole genome duplication. One exception is ogn for which duplicate copies were identified in fish genomes. The ogn promoter sequence and in vitro mesenchymal stem cell (MSC) cultures suggest the duplicate ogn genes acquired divergent functions. In gilthead sea bream (Sparus aurata) ogn1 was up-regulated during osteoblast and myocyte differentiation in vitro, while ogn2 was severely down-regulated during bone-derived MSCs differentiation into adipocytes in vitro. Conclusions Overall, the phylogenetic analysis indicates that the SLRP III family in vertebrates has been under conservative evolutionary pressure. The retention of the ogn gene duplicates in teleosts was linked with the acquisition of different functions. The acquisition by OGN of functions other than that of a bone ECM protein occurred early in the vertebrate lineage.
- Thermal imprinting during embryogenesis modifies skin repair in juvenile European sea bass (Dicentrarchus labrax)Publication . Mateus, Ana; Costa, Rita; Sadoul, Bastien; Bégout, Marie-Laure; Cousin, Xavier; Canario, Adelino; Power, DeborahFish skin is a multifunctional tissue that develops during embryogenesis, a developmental stage highly suscep-tible to epigenetic marks. In this study, the impact of egg incubation temperature on the regeneration of a cutaneous wound caused by scale removal in juvenile European sea bass was evaluated. Sea bass eggs were incubated at 11, 13.5 and 16 degrees C until hatching and then were reared at a common temperature until 9 months when the skin was damaged and sampled at 0, 1 and 3 days after scale removal and compared to the intact skin from the other flank. Skin damage elicited an immediate significant (p < 0.001) up-regulation of pcna in fish from eggs incubated at higher temperatures. In fish from eggs incubated at 11 C there was a significant (p < 0.001) up-regulation of krt2 compared to fish from higher thermal backgrounds 1 day after skin damage. Damaged epidermis was regenerated after 3 days in all fish irrespective of the thermal background, but in fish from eggs incubated at 11 C the epidermis was significantly (p < 0.01) thinner compared to other groups, had less goblet cells and less melanomacrophages. The thickness of the dermis increased during regeneration of wounded skin irrespective of the thermal background and by 3 days was significantly (p < 0.01) thicker than the dermis from the intact flank. The expression of genes for ECM remodelling (mmp9, colX alpha, col1 alpha 1, sparc, and angptl2b) and innate immunity (lyg1, lalba, sod1, csf-1r and ppar gamma) changed during regeneration but were not affected by egg thermal regime. Overall, the results indicate that thermal imprinting of eggs modifies the damage-repair response in juvenile sea bass skin.
- GABA receptors in the olfactory epithelium of the gilthead seabream (Sparus aurata)Publication . Costa, Rita; Velez, Zélia; Hubbard, PeterExposure to high PCO2/low pH seawater induces behavioural alterations in fish; a possible explanation for this is a reversal of Cl-/HCO3- currents through GABAA receptors (the GABAA receptor theory). However, the main evidence for this is that gabazine, a GABAA receptor antagonist, reverses these effects when applied to the water, assuming that exposure to systems other than the CNS would be without effect. Here, we show the expression of both metabotropic and ionotropic GABA receptors, and the presence of GABAA receptor protein, in the olfactory epithelium of gilthead seabream. Furthermore, exposure of the olfactory epithelium to muscimol (a specific GABAA receptor agonist) increases or decreases the apparent olfactory sensitivity to some odorants. Thus, although the exact function of GABAA receptors in the olfactory epithelium is not yet clear, this may complicate the interpretation of studies wherein water-borne gabazine is used to reverse the effects of high CO2 levels on olfactory-driven behaviour in fish.
- Independent effects of seawater pH and high P-CO2 on olfactory sensitivity in fish: possible role of carbonic anhydrasePublication . Velez, Zélia; Costa, Rita; Wang, Wenjing; Hubbard, PeterOcean acidificationmay alter olfactory-driven behaviour in fish by direct effects on the peripheral olfactory system; olfactory sensitivity is reduced in CO2-acidified seawater. The current study tested whether this is due to elevated P-CO2 or the consequent reduction in seawater pH and, if the former, the possible involvement of carbonic anhydrase, the enzyme responsible for the hydration of CO2 and production of carbonic acid. Olfactory sensitivity to amino acids was assessed by extracellularmulti-unit recording from the olfactory nerve of the gilthead seabream (Sparus aurata L.) in normal seawater (pH similar to 8.2), and after acute exposure to acidified seawater (pH similar to 7.7) but normal P-CO2 (similar to 340 mu atm) or to high P-CO2 seawater (similar to 1400 mu atm) at normal pH (similar to 8.2). Reduced pH in the absence of elevated P-CO2 caused a reduction in olfactory sensitivity to L-serine, L-leucine, L-arginine and L-glutamine, but not L-glutamic acid. Increased P-CO2 in the absence of changes in pH caused reduced olfactory sensitivity to L-serine, L-leucine and L-arginine, including increases in their threshold of detection, but had no effect on sensitivity to L-glutamine and L-glutamic acid. Inclusion of 1 mmol l(-1) acetazolamide (a membrane-permeant inhibitor of carbonic anhydrase) in the seawater reversed the inhibition of olfactory sensitivity to L-serine caused by high P-CO2. Ocean acidification may reduce olfactory sensitivity by reductions in seawater pH and intracellular pH (of olfactory receptor neurones); the former by reducing odorant-receptor affinity, and the latter by reducing the efficiency of olfactory transduction. The physiological role of carbonic anhydrase in the olfactory receptor neurones remains to be explored.
- Persistence of the ABCC6 genes and the emergence of the bony skeleton in vertebratesPublication . Parreira, Bruna; Cardoso, João; Costa, Rita; Couto, Ana Rita; Bruges-Armas, Jacome; Power, Deborah M.The ATP-binding cassette transporter 6 (ABCC6) gene encodes a cellular transmembrane protein transporter (MRP6) that is involved in the regulation of tissue calcification in mammals. Mutations in ABCC6 are associated with human ectopic calcification disorders. To gain insight into its evolution and involvement in tissue calcification we conducted a comparative analysis of the ABCC6 gene and the related gene ABCC1 from invertebrates to vertebrates where a bony endoskeleton first evolved. Taking into consideration the role of ABCC6 in ectopic calcification of human skin we analysed the involvement of both genes in the regeneration of scales, mineralized structures that develop in fish skin. The ABCC6 gene was only found in bony vertebrate genomes and was absent from Elasmobranchs, Agnatha and from invertebrates. In teleost fish the abcc6 gene duplicated but the two genes persisted only in some teleost genomes. Six disease causing amino acid mutations in human MRP6 are a normal feature of abcc6 in fish, suggesting they do not have a deleterious effect on the protein. After scale removal the abcc6 (5 and 10 days) and abcc1 (10 days) gene expression was up-regulated relative to the intact control skin and this coincided with a time of intense scale mineralization.
- Cartilage acidic protein a novel therapeutic factor to improve skin damage repair?Publication . Félix, Rute; Anjos, Liliana; Costa, Rita; Letsiou, Sophia; Power, Deborah MaryFish skin has been gaining attention due to its efficacy as a human-wound-treatment product and to identify factors promoting its enhanced action. Skin fibroblasts have a central role in maintaining skin integrity and secrete extra cellular matrix (ECM) proteins, growth factors and cytokines to rapidly repair lesions and prevent further damage or infection. The effects on scratch repair of the ubiquitous but poorly characterized ECM protein, cartilage acidic protein 1 (CRTAC1), from piscine and human sources were compared using a zebrafish SJD.1 primary fibroblast cell line. A classic in vitro cell scratch assay, immunofluorescence, biosensor and gene expression analysis were used. Our results demonstrated that the duplicate sea bass Crtac1a and Crtac1b proteins and human CRTAC-1A all promoted SJD.1 primary fibroblast migration in a classic scratch assay and in an electric cell impedance sensing assay. The immunofluorescence analysis revealed that CRTAC1 enhanced cell migration was most likely caused by actin-driven cytoskeletal changes and the cellular transcriptional response was most affected in the early stage (6 h) of scratch repair. In summary, our results suggest that CRTAC1 may be an important factor in fish skin promoting damage repair.
- Skin and scale regeneration after mechanical damage in a teleostPublication . Costa, Rita; Power, DeborahSkin wound healing has been widely studied in mammalian models but the information on teleost cutaneous healing is sparse and frequently considered in the context of viral or bacterial infections or parasitic infestations in aquaculture. In the present study a detailed time course (0 h, 6 h, 1, 2, 3 and 4 days) coupled to morphology and gene expression analysis revealed rapid regeneration of skin without scarring in a marine teleost after a superficial wound caused by the loss of a large area of scales. The integrity of the integument, as assessed by quantification of extracellular matrix (ECM) gene transcripts (fn1a, colIα1, colVα2, colXα1, ogn1, ogn2, crtac1a, cyr61, pcna, krt2 and mmp9) was restored within 2 days. Epithelial-mesenchyme interactions assessed by expression of edar and shh were associated with epidermal closure, the re-establishment of the basement membrane and also scale eruption. Histological observations suggested tissue re-epithelialization was independent of inflammation and that transcripts representing the humoral and cellular elements of the immune response (mpo, cyba and csf1r, cd48 and cd200) were modulated in the early stages of sea bream (Sparus aurata) skin repair after injury. Overall, the results indicate that after superficial skin damage tissue reconstitution started immediately with re-epithelialization, followed by ECM deposition and finally tissue maturation, indicating that in the skin regenerative process, reconstitution of the physical barrier was the priority over other integument functions, including immune surveillance.
- Ocean acidification affects the expression of neuroplasticity and neuromodulation markers in seabreamPublication . A Costa, Rita; Olvera, Aurora; Power, Deborah Mary; Velez, ZéliaA possible explanation for acidification-induced changes in fish behaviour is that acidification interferes with neurogenesis and modifies the plasticity of neuronal circuitry in the brain. We tested the effects on the olfactory system and brain of gilthead seabream (Sparus aurata) to 4 weeks' exposure to ocean acidification (OA). Olfactory epithelium (OE) morphology changed shortly after OA exposure and persisted over the 4 weeks. Expression of genes related to olfactory transduction, neuronal excitability, synaptic plasticity, GABAergic innervation, and cell proliferation were unchanged in the OE and olfactory bulb (OB) after 4 weeks' exposure. Short-term changes in the ionic content of plasma and extradural fluid (EDF) returned to control levels after 4 weeks' exposure, except for [Cl-], which remained elevated. This suggests that, in general, there is an early physiological response to OA and by 4 weeks a new homeostatic status is achieved. However, expression of genes involved in proliferation, differentiation and survival of undifferentiated neurons were modified in the brain. In the same brain areas, expression of thyroid hormone signalling genes was altered suggesting modifications in the thyroid-system may be linked to the changes in neuroplasticity and neurogenesis. Overall, the results of the current study are consistent with and effect of OA on neuroplasticity.
- Temperature responsiveness of gilthead sea bream bone; an in vitro and in vivo approachPublication . Riera-Heredia, Natalia; Martins, Rute; Patricia Mateus, Ana; Costa, Rita; Gisbert, Enric; Navarro, Isabel; Gutierrez, Joaquim; Power, Deborah M.; Capilla, EncarnacionThis study aimed to characterize the molecules involved in osteogenesis in seabream and establish using in vitro/in vivo approaches the responsiveness of selected key genes to temperature. The impact of a temperature drop from 23 to 13 degrees C was evaluated in juvenile fish thermally imprinted during embryogenesis. Both, in vitro/in vivo, Fib1a, appeared important in the first stages of bone formation, and Col1A1, ON and OP, in regulating matrix production and mineralization. OCN mRNA levels were up-regulated in the final larval stages when mineralization was more intense. Moreover, temperature-dependent differential gene expression was observed, with lower transcript levels in the larvae at 18 degrees C relative to those at 22 degrees C, suggesting bone formation was enhanced in the latter group. Results revealed that thermal imprinting affected the long-term regulation of osteogenesis. Specifically, juveniles under the low and low-to-high-temperature regimes had reduced levels of OCN when challenged, indicative of impaired bone development. In contrast, gene expression in fish from the high and high-to-low-temperature treatments was unchanged, suggesting imprinting may have a protective effect. Overall, the present study revealed that thermal imprinting modulates bone development in seabream larvae, and demonstrated the utility of the in vitro MSC culture as a reliable tool to investigate fish osteogenesis.
- The impact of egg thermal regimes on the response to food deprivation and refeeding in juvenile European sea bass (Dicentrarchus labrax)Publication . Mateus, Ana; Costa, Rita; Jiménez, Javier; Sadoul, Bastien; Bégout, Marie Laure; Cousin, Xavier; Canario, Adelino; Power, DeborahFish are ectotherms and this means they are highly vulnerable to changes in ambient temperature, particularly during early developmental stages when temperature can induce persistent effects on phenotypic traits. In this study, the effect of egg incubation temperature on the response of juvenile European sea bass (Dicentrarchus labrax) to food deprivation and recovery after refeeding was assessed. Eggs were incubated at 11, 13.5 and 16 & DEG;C until hatching and then were reared at a common temperature until 9 months when fish were deprived of food for one week. The recovery from food deprivation was evaluated at 10 h and 2 days post-refeeding. Food deprivation in fish from eggs incubated at the highest temperature (16 & DEG;C) compared to 11 and 13.5 & DEG;C exhibited the most morphological and metabolic changes in the liver and foregut. Liver metabolism was changed as revealed by the significant reduction in lipid area and the increased number of hepatocyte nuclei. Foregut atrophy was coupled to a significant up-regulation of transcripts associated with gluconeogenesis (pck1) and peptide absorption (pept1). A modified metabolic response to the fast-refeed regime was revealed by the significantly decreased levels of plasma lactate, which may result from up-regulation of transcripts of the thyroid axis, deiodinase 2 (dio2) in the foregut. Fish incubated as eggs at a lower temperature (11 & DEG;C) exhibited less changes following the fast-refeed regime. Food deprivation did not significantly modify the morphology of the foregut and the liver parenchyma recovered sooner in fish from the 11 & DEG;C egg thermal regime compared to fish from the other thermal regimes following refeeding. The latter group of fish had a temporary stimulation of the GH-IGF axis with significant up-regulation of liver insulin-like growth factor I and II (igf-1 and igf-2) after fasting. The liver parenchyma of fish from the 13.5 & DEG;C egg thermal regime (the standard temperature of the hatchery stage) did not recover by the end of the experiment and transcripts of catalase (cat), encoding an antioxidant enzyme, were significantly downregulated compared to fish from the other egg thermal regimes. Our results suggest that thermal imprinting at the egg stage in European sea bass modified the juvenile metabolic response to food deprivation and the recovery response when feeding was resumed.