Browsing by Author "Modesto, Teresa"
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- 16.P15. Branchial osmoregulatory response to salinity challenge in the Lusitanian toadfishPublication . Guerreiro, Pedro M; Laiz-Carrión, R.; Haond, Christophe; Modesto, Teresa; Fuentes, Juan; Mancera, J. M.; Canario, Adelino V. M.Halobatrachus didactylus, a marine teleost found in coastal lagoons and river estuaries is often exposed to important salinity changes. Despite its aglomerular kidney, it is able to survive in hypo-osmotic environments, likely via compensatory actions from gills and intestine. We aimed at evaluating the response of the branchial tissue of H. didactylus to salinity changes.
- 17a,20ß,21-trihydroxy-4-pregnen-3-one: the probable maturation-inducing steroid of the lusitanian toadfishPublication . Modesto, Teresa; Canario, Adelino V. M.17,20 ,21-Trihydroxy-4-pregnen-3-one (17,20 ,21-P) was identified as the major metabolite of incubations of Lusitanian toadfish Halobatrachus didactylus ovarian follicles with [3H]-17hydroxyprogesterone. The potency of several steroids in inducing germinal vesicle breakdown of follicle-enclosed oocytes of Lusitanian toadfish was systematically examined by using an in vitro germinal vesicle breakdown (GVBD) bioassay. 17,20 -Dihydroxy-4-pregnen-3-one (17,20 -P) and 17,20 ,21-P, two confirmed maturation-inducing steroids (MIS) in teleosts, were the most potent in inducing GVBD with ED50s ranging between 9 and 271 nM. Structure-activity relationships followed similar patterns to what has been observed in similar bioassays, i.e. a vital requirement for 17- and 20 -hydroxyl groups in C21 steroids and a reduction in activity of 14 and 5–6%, respectively, for 5-pregnene and 5 -pregnanes compared to 4-pregnenes. Corticosteroids, testosterone and 17 -oestradiol were ineffective. Folliculated oocytes stimulated by pituitary homogenate produced 17,20 ,21-P from endogenous substrates in amounts one order of magnitude higher than 17,20 -P. These results strongly support the hypothesis that 17,20 ,21-P is the likely MIS in this species.
- Alloparental behavior in the highly vocal Lusitanian toadfishPublication . Ramos, Andreia; Fonseca, Paulo; Modesto, Teresa; Almada, V. C.; Amorim, Maria ClaraIn many fish species in which males guard nests with their eggs, parental care directed to genetically unrelated offspring may arise for example from nest takeovers or cuckoldry. Lusitanian toadfish (Halobatrachus didactylus) has exclusive male parental care and face intensive nest competition during the breeding season that may lead to care of foster eggs. Males of this species use visual displays and sounds when defending their nests frequently resulting in expulsion of the intruder without escalated confrontation. In this study we intended to investigate the existence of alloparental care in Lusitanian toadfish, a behavior whose adaptive significance is still poorly understood. Fish were randomly assigned to three different treatments: parental males in nests with their eggs, parental males with foster eggs and parental males without eggs. Nests with eggs with no nest holder or with females were used as controls. We performed three territorial intrusions over periods of 15 days and observed the acoustic and visual behaviors of residents and intruders. Egg survival was tallied from nests' photographs in all groups. Circulating steroid levels were measured in the three test groups and in another set of non-manipulated males. There were no differences in acoustic and visual territorial defense behaviors among treatments. Egg survival was similar between males (parental and alloparental) and significantly higher than in nests with no nest-tender. Females presented intermediate egg survival. All groups presented similar levels of testosterone and alloparental males showed higher 11-ketotestosterone levels but within the range of levels observed in non-manipulated males. Cortisol levels were similar in all male groups suggesting that experiments did not increase fish stress. The present results suggest the existence of alloparental care in this species.
- Anatomy of the olfactory system and potential role for chemical communication in the sound‐producing lusitanian toadfish, halobatrachus didactylusPublication . Modesto, Teresa; Gregório, Beatriz Neves; Marcelino, Gonçalo; Marquet, Nathalie; Costa, Rita; Guerreiro da Costa Guerreiro, Pedro Miguel; Velez, Zélia; Hubbard, PeterThe current study investigated the structure and function of the olfactory system of the Lusitanian toadfish, Halobatrachus didactylus, using histology and electrophysiology (electro-olfactogram [EOG]), respectively. The olfactory system consists of a digitated anterior peduncle, of unknown function, containing the inhalant nostril. This then leads to a U-shaped olfactory chamber with the olfactory epithelium-identified by G(alpha olf)-immunoreactivity-on the ventral surface. A large lacrimal sac is connected to this tube and is likely involved in generating water movement through the olfactory chamber (this species is largely sedentary). The exhalent nostril lies by the eye and is preceded by a bicuspid valve to ensure one-way flow of water. As do other teleosts, H. didactylus had olfactory sensitivity to amino acids and bile acids. Large-amplitude EOG responses were evoked by fluid from the anterior and posterior testicular accessory glands, and bile and intestinal fluids. Anterior gland and intestinal fluids from reproductive males were significantly more potent than those from non-reproductive males. Male urine and skin mucus proved to be the least potent body fluids tested. These results suggest that chemical communication-as well as acoustic communication-may be important in the reproduction of this species and that this may be mediated by the accessory glands and intestinal fluid.
- Boat noise impacts Lusitanian toadfish breeding males and reproductive outcomePublication . Amorim, M. Clara P.; Vieira, Manuel; Meireles, Gabriela; Novais, Sara C.; Lemos, Marco F. L.; Modesto, Teresa; Alves, Daniel; Zuazu, Ana; Lopes, Ana F.; Matos, André B.; Fonseca, Paulo J.Anthropogenic noise is a growing threat to marine organisms, including fish. Yet very few studies have addressed the impact of anthropogenic noise on fish reproduction, especially in situ. In this study, we investigated the impacts of boat noise exposure in the reproductive success of wild Lusitanian toadfish (Halobatrachus didactylus), a species that relies on advertisement calls for mate attraction, using behavioural, physiological and reproductive endpoints. Two sets of artificial nests were deployed in the Tagus estuary and exposed to either ambient sound or boat noise during their breeding season. Toadfish males spontaneously used these nests to breed. We inspected nests for occupation and the presence of eggs in six spring low tides (in two years) and assessed male vocal activity and stress responses. Boat noise did not affect nest occupation by males but impacted reproductive success by decreasing the likelihood of receiv-ing eggs, decreasing the number of live eggs and increasing the number of dead eggs, compared to control males. Treat-ment males also showed depressed vocal activity and slightly higher cortisol levels. The assessment of oxidative stress and energy metabolism-related biomarkers revealed no oxidative damage in noise exposed males despite having lower antioxidant responses and pointed towards a decrease in the activity levels of energy metabolism-related biomarkers. These results suggest that males exposed to boat noise depressed their metabolism and their activity (such as parental care and mate attraction) to cope with an acoustic stressor, consistent with a freezing defensive response/behaviour. Together, our study demonstrates that boat noise has severe impacts on reproductive fitness in Lusitanian toadfish. We argue that, at least fishes that cannot easily avoid noise sources due to their dependence on specific spawning sites, may incur in significant direct fitness costs due to chronic noise exposure.
- Effect of dietary l-glutamine supplementation on the intestinal physiology and growth during Solea senegalensis larval developmentPublication . Matias, Ana Catarina; Viegas, Ana Rita; Couto, Ana; Lourenço-Marques, Cátia; Aragão, Cláudia; Castanho, Sara; Gamboa, Margarida; Candeias-Mendes, Ana; Soares, Florbela; Modesto, Teresa; Pousão-Ferreira, Pedro; Ribeiro, LauraThe maturation of the intestinal digestive and absorptive functions might limit the amount of absorbed nutrients to fulfil the high requirements of the fast-growing marine fish larva. Glutamine (Gln) has been described to improve intestinal epithelium functions, due to its involvement in energy metabolism and protein synthesis. The purpose of this study was to evaluate dietary 0.2% Gln supplementation on aspects of intestinal physiology, protein metabolism and growth -related genes expression in Senegalese sole larvae. Experiment was carried out between 12 and 33 days post hatching (DPH) and fish were divided into two experimental groups, one fed Artemia spp. (CTRL) and the other fed Artemia spp. supplemented with Gln (GLN). GLN diet had two times more Gln than the CTRL diet. Samples were collected at 15, 19, 26 and 33 DPH for biometry, histology, and digestive enzymes activity, and at 33 DPH for gene expression, protein metabolism and AA content determination. Growth was significantly higher for Senegalese sole fed GLN diet, supported by differences on protein metabolism and growth -related gene expression. Slight differences were observed between treatments regarding the intestinal physiology. Overall, GLN diet seems to be directed to enhance protein metabolism leading to higher larval growth.
- Hormonal control of swimbladder sonic muscle dimorphism in the Lusitanian toadfish, Halobatrachus didactylusPublication . Modesto, Teresa; Canario, Adelino V. M.The swimbladder and associated sonic muscle of the Lusitanian toadfish Halobatrachus didactylus increase in size throughout life and are, respectively, 25% and 30% larger in type I (nest-holder) males than females, which may generate sexual differences in sound production. Sexual dimorphism in swimbladder is also evident in the morphological features of sonic muscle fibers. During the breeding season, type I males have smaller myofibril contracting zones surrounded by larger sarcoplasm areas compared with females, possibly an adaptation to speed and fatigue resistance for the production of long mating calls. Type II (floater) males show characteristics that are intermediate, but statistically not significantly different, between type I males and females. Six weeks after castration and androgen (testosterone and 11- ketotestosterone) replacement in type I and type II males there were no alterations either in swimbladder mass or fiber morphology. However, 17b-estradiol induced a significant decrease in swimbladder mass and sarcoplasm area/myofibril area ratio. Six months after castration there was a clear reduction in the seasonal swimbladder hypertrophy in males and induction of sonic fiber morphological characteristics that resemble those occurring in females (low sarcoplasm area/myofibril area ratio). These results suggest that testicular factors are required to initiate sonic muscle hypertrophy and type I sonic fiber phenotype in H. didactylus, but a specific involvement of androgens has not been completely clarified.
- Molecular characterization and expression pattern of zona pellucida proteins in gilthead seabream (Sparus aurata)Publication . Modig, Carina; Modesto, Teresa; Canario, Adelino V. M.; Cerda, J.; Hofsten, Jonas von; Olsson, P. E.The developing oocyte is surrounded by an acellular envelope that is composed of 2–4 isoforms of zona pellucida (ZP) proteins. The ZP proteins comprise the ZP1, ZP2, ZP3, and ZPX isoforms. While ZP1 (ZPB) and ZP3 (ZPC) are present in all species, ZP2 (ZPA) is not found in teleost fish and ZPX is not found in mammals. In the present study, we identify and characterize the ZP1, ZP3 and ZPX isoforms of gilthead seabream. Furthermore, by analyzing the conserved domains, which include the external hydrophobic patch and the internal hydrophobic patch, we show that ZP2 and ZPX are closely related isoforms. ZP proteins are synthesized in either the liver or ovary of most teleosts. Only in rainbow trout has it been shown that zp3 has dual transcription sites. In gilthead seabream, all four mRNA isoforms are transcribed in both the liver and ovary, with zp1a, zp1b, and zp3 being highly expressed in the liver, and zpx being primarily expressed in the ovary. However, determination of the ZP proteins in plasma showed high levels of ZP1b, ZP3, and ZPX, with low or non-detectable levels of ZP1a. In similarity to other teleost ZPs, the hepatic transcription of all four ZP isoforms is under estrogenic control. Previously, we have shown that cortisol can potentiate estrogen-induced ZP synthesis in salmonids, and now we show that this is not the case in the gilthead seabream. The present study shows for the first time the endocrine regulation of a teleost ZPX isoform, and demonstrates the dual-organ transcriptional activities of all the ZP proteins in one species.
- Morphometric changes and sex steroid levels during the annual reproductive cycle of the Lusitanian toadfish, Halobatrachus didactylusPublication . Modesto, Teresa; Canario, Adelino V. M.The Lusitanian toadfish has group synchronous oocytes, which grow from November until June–July when they are released probably as a single batch. Blood plasma levels of estradiol-17b (E2) and testosterone (T) increase during vitellogenesis and drop rapidly during final maturation and ovulation, when 17,20b, 21-trihydroxy-4-pregnen-3-one (17,20b,21-P) levels increase. The male reproductive apparatus is composed of paired testes and multichambered accessory glands, which secrete mucosubstances and are connected to the spermatic duct. Changes in the gonadosomatic index of males paralleled the females but started to drop slightly earlier. The swimbladder and accessory glands also underwent important seasonal changes in weight reaching a maximum at spawning. T, 11-ketotestosterone (11-KT) and 17,20a-dihydroxy-4-pregnen-3-one (17,20a-P) were generally low except for a sharp peak in June. 17,20b,21-P also peaked in June and then declined slowly. 17,20b-dihydroxy-4-pregnen-3-one (17,20a-P) was undetectable in males and females. As with other species of the family two types of males were identified: type I males with smaller testes (ca. 7-fold) and larger accessory glands (ca. 3-fold) and swimbladders than type II. Type I males also had significantly higher (ca. 6- fold) 11-KT levels than type II males. This suggests a role for 11-KT in the development of structures important for reproductive behaviour.
- A PTH/PTHrP receptor antagonist blocks the hypercalcemic response to estradiol-17bPublication . Fuentes, J.; Guerreiro, P. M.; Modesto, Teresa; Rotllant, J.; Canario, Adelino V. M.; Power, DeborahEstradiol (E2) increases circulating calcium and phosphate levels in fish, thus acting as a hypercalcemic and hyperphosphatemic factor during periods of high calcium requirements, such as during vitellogenesis. Since parathyroid hormone (PTH)-related protein (PTHrP) has been shown to be calciotropic in fish, we hypothesized that the two hormones could be mediating the same process. Sea bream (Sparus auratus) juveniles receiving a single intraperitoneal injection of piscine PTHrP(1-34) showed an elevation in calcium plasma levels within 24 h. In contrast, injections of the PTH/PTHrP receptor antagonist PTHrP(7-34) decreased circulating levels of calcium in the same period. Intraperitoneal implants of estradiol-17 (E2; 10 g/g) evoked significant increases of circulating plasma levels of calcium and phosphorus and a sustained increases of circulating plasma levels of PTHrP. However, a combined treatment of E2 and PTHrP(7-34) evoked a markedly lower calcium response compared with E2 alone. We conclude that PTHrP or a related peptide that binds the PTH/PTHrP receptor mediates, at least in part, the hypercalcemic effect of E2 in calcium and phosphate balance in fish.