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Duplication of Dio3 genes in teleost fish and their divergent expression in skin during flatfish metamorphosis

Publication . Alves, Ricardo; Cardoso, João CR; Harboe, T.; Martins, Rute; Manchado, M.; Norberg, B.; Power, Louise

Deiodinase 3 (Dio3) plays an essential role during early development in vertebrates by controlling tissue thyroid hormone (TH) availability. The Atlantic halibut (Hippoglossus hippoglossus) possesses duplicate dio3 genes (dio3a and dio3b). Expression analysis indicates that dio3b levels change in abocular skin during metamorphosis and this suggests that this enzyme is associated with the divergent development of larval skin to the juvenile phenotype. In larvae exposed to MMI, a chemical that inhibits TH production, expression of dio3b in ocular skin is significantly up-regulated suggesting that THs normally modulate this genes expression during this developmental event. The molecular basis for divergent dio3a and dio3b expression and responsiveness to MMI treatment is explained by the multiple conserved TREs in the proximal promoter region of teleost dio3b and their absence from the promoter of dio3a. We propose that the divergent expression of dio3 in ocular and abocular skin during halibut metamorphosis contributes to the asymmetric pigment development in response to THs. (C) 2017 Elsevier Inc. All rights reserved.

2017-05Journal article

Restricted

Compounded perturbations in coastal areas: contrasting responses to nutrient enrichment and the regime of storm-related disturbance depend on life-history traits

Publication . Bertocci, Iacopo; DOMINGUEZ GODINO, JORGE ANTONIO; Freitas, Cristiano; Incera, Monica; Bio, Ana; Dominguez, Rula

1. Natural systems are exposed to compounded perturbations, whose changes in temporal variance can be as important as those in mean intensity for shaping the structure of assemblages. Specifically, climate-related physical disturbances and nutrient inputs due to natural and/or anthropogenic activities occur concomitantly, but experimental tests of the simultaneous effects of changes in the regime of more than one perturbation are generally lacking. Filling this gap is the key to understand ecological responses of natural assemblages to climate-related change in the intensity and temporal patterning of physical disturbance combined with other global stressors. 2. Responses to factorial manipulations of nutrient enrichment, mean intensity and temporal variability in storm-like mechanical disturbance were examined, using benthic assemblages of tide-pools as model system. 3. Response variables were mean abundance values and temporal variances of taxa with different life-traits. Consistent negative effects of disturbance intensity were observed for the mean cover of long-living taxa (algal canopies and the polychaete Sabellaria alveolata), whose temporal fluctuations were also reduced by more severe mechanical stress. More resilient taxa (ephemeral algae, mostly green of the genus Ulva) increased under enriched conditions, particularly when low-intensity events were irregularly applied over time. Opposite effects of disturbance intensity depending on nutrient availability occurred on filamentous algae (e.g. red of the genus Ceramium). This was probably due to the fact that, although nutrient enrichment stimulated the abundance of both algal groups, when this condition was combined with relatively mild physical disturbance the competitively superior ephemeral green algae tended to become dominant over filamentous red algae. The same did not occur under high intensity of disturbance since it likely damaged large, foliose fronds of Ulva-like forms more than small, filamentous fronds of Ceramium-like forms. Grazers were positively affected by nutrients, likely responding indirectly to more food available. 4. A direct relationship between the mean abundance of most organisms and their temporal fluctuations was documented. However, all organisms persisted throughout the study, even under experimental conditions associated to the largest temporal variation in their abundance, likely due to their ability to resist to/quickly recover from, the applied perturbations. Therefore, in systems with great recovery abilities of dominant organisms (e.g. rocky intertidal, grasslands), effects of traits of the regime of disturbance and nutrient enrichment may modulate the fluctuations of populations not through the elimination and substitution of species, but through changes in relative abundances of the same species. This contrasts with the theory that temporal variation in abundance would be directly related to the risk of local extinction. Present findings enable more accurate predictions of the consequences of climatic and non-climatic scenarios on the biodiversity of marine and terrestrial systems sharing analogous functional traits of organisms. Future more intense physical disturbances are expected to exert negative effects on slow-growing/recovering species (e.g. habitat-formers) irrespectively of the temporal patterning of the same disturbances and nutrient inputs. On the contrary, more resistant species (e.g. encrusting algae on rocky shores or below-ground vegetation in grasslands) are expected to benefit from intense physical disturbance. Species whose abundance is more directly related to the availability of nutrients (e.g. filamentous and ephemeral algae or herbs) are expected to generally increase under enriched conditions, but their ability to eventually become dominant would depend on their ability to grow fast and attain cover large enough to overwhelm any possible control of concomitant disturbance intensity on their abundance. If, such as in the present examined system, virtually all organisms can persist, over the temporal scale of the experiment, under any combination of physical disturbance and nutrient availability, the resulting overall diversity is not predicted to change drastically. Nevertheless, low-intensity events evenly distributed and high-intensity events irregularly distributed appear as the conditions supporting the highest richness of taxa, independently of the availability of nutrients.

2017-05Journal article

Open Access

Early transition to microdiets improves growth, reproductive performance and reduces skeletal anomalies in Zebrafish (Danio rerio)

Publication . Martins, Gil; Diogo, Patricia; Pinto, Wilson; Gavaia, Paulo

Zebrafish is a model species with a high variability of feeding regimes among fish facilities. The use of live feeds for early life stages is a common practice, and few studies have focused early weaning into microdiets. The lack of standardized feeding protocols among research facilities promotes discrepancies in biological performances, and few studies relate dietary regimes to zebrafish development. The objective of this work was to assess the effect of an early transition into microdiets in zebrafish development by evaluating growth, survival, reproductive performance, and skeletal anomalies. These parameters were assessed in one group exclusively fed on Artemia nauplii and two groups fed on microdiets (commercial and experimental). Results showed that an early weaning with the two microdiets significantly improved zebrafish growth and reproductive performance, while a decrease in incidence of vertebral column anomalies was observed. A high survival was also maintained in fish fed microdiets at an early developmental stage when comparing to exclusive Artemia nauplii feeding. In conclusion, early weaning with high quality microdiets is beneficial for zebrafish growth, reproductive performance, and skeletal development, contributing to the standardization of zebrafish husbandry practices.

2019Journal article

Open Access

Prenylated chalcone 2 ccts as an antimitotic agent and enhances the chemosensitivity of tumor cells to paclitaxel

Publication . Fonseca, Joana; Marques, Sandra; Silva, PMA; Patrícia M.A. Silva; Brandão, Pedro; Cidade, Honorina; Pinto, Madalena M.; Bousbaa, Hassan

We previously reported that prenylated chalcone 2 (PC2), the O-prenyl derivative (2) of 2'-hydroxy-3,4,4',5,6'-pentamethoxychalcone (1), induced cytotoxicity of tumor cells via disruption of p53-MDM2 interaction. However, the cellular changes through which PC2 exerts its cytotoxic activity and its antitumor potential, remain to be addressed. In the present work, we aimed to (i) characterize the effect of PC2 on mitotic progression and the underlying mechanism; and to (ii) explore this information to evaluate its ability to sensitize tumor cells to paclitaxel in a combination regimen. PC2 was able to arrest breast adenocarcinoma MCF-7 and non-small cell lung cancer NCI-H460 cells in mitosis. All mitosis-arrested cells showed collapsed mitotic spindles with randomly distributed chromosomes, and activated spindle assembly checkpoint. Live-cell imaging revealed that the compound induced a prolonged delay (up to 14 h) in mitosis, culminating in massive cell death by blebbing. Importantly, PC2 in combination with paclitaxel enhanced the effect on cell growth inhibition as determined by cell viability and proliferation assays. Our findings demonstrate that the cytotoxicity induced by PC2 is mediated through antimitotic activity as a result of mitotic spindle damage. The enhancement effects of PC2 on chemosensitivity of cancer cells to paclitaxel encourage further validation of the clinical potential of this combination.

2016-08Journal article

Open Access

European Sea Bass (Dicentrarchus labrax) immune status and disease resistance are impaired by arginine dietary supplementation

Publication . Azeredo, Rita; Perez-Sanchez, Jaume; Sitja-Bobadilla, Ariadna; Fouz, Belen; Tort, Lluis; Aragao, Claudia; Oliva-Teles, Aires; Costas, Benjamin

Infectious diseases and fish feeds management are probably the major expenses in the aquaculture business. Hence, it is a priority to define sustainable strategies which simultaneously avoid therapeutic procedures and reinforce fish immunity. Currently, one preferred approach is the use of immunostimulants which can be supplemented to the fish diets. Arginine is a versatile amino acid with important mechanisms closely related to the immune response. Aiming at finding out how arginine affects the innate immune status or improve disease resistance of European seabass (Dicentrarchus labrax) against vibriosis, fish were fed two arginine-supplemented diets (1% and 2% arginine supplementation). A third diet meeting arginine requirement level for seabass served as control diet. Following 15 or 29 days of feeding, fish were sampled for blood, spleen and gut to assess cell-mediated immune parameters and immune-related gene expression. At the same time, fish from each dietary group were challenged against Vibrio anguillarum and survival was monitored. Cell-mediated immune parameters such as the extracellular superoxide and nitric oxide decreased in fish fed arginine-supplemented diets. Interleukins and immune-cell marker transcripts were down-regulated by the highest supplementation level. Disease resistance data were in accordance with a generally depressed immune status, with increased susceptibility to vibriosis in fish fed arginine supplemented diets. Altogether, these results suggest a general inhibitory effect of arginine on the immune defences and disease resistance of European seabass. Still, further research will certainly clarify arginine immunomodulation pathways thereby allowing the validation of its potential as a prophylactic strategy.

2015-10Journal article

Open Access