Browsing by Author "Cerda, J."
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- Evidence for involvement of chemical communication in reproduction of the eel Anguilla anguillaPublication . Huertas, Mar; Hubbard, Peter; Scott, A. P.; Canario, Adelino V. M.; Cerda, J.The European eel (Anguilla anguilla) is characterised by long transoceanic migration to spawning sites during which the fish become sexually mature. A previous study demonstrated that mature eels may induce physiological and behavioural changes in immature conspecifics. This raises the possibility that chemical communication may be involved in this process. The aim of the current study was, therefore, to assess the olfactory sensitivity of eels to conspecific-derived odours and to establish whether these odours differ according to sex and/or state of maturity.
- Isolation of a novel aquaglyceroporin from a marine teleost (Sparus auratus) Function and tissue distributionPublication . Santos, C. R. A.; Estêvão, Dulce; Fuentes, J.; Cardoso, João CR; Fabra, Mercedes; Passos, A. L.; Detmers, F. J.; Deen, P. M. T.; Cerda, J.; Power, DeborahThe aquaporins (formerly called the major intrinsic protein family) are transmembrane channel proteins. The family includes the CHIP group, which are functionally characterised as water channels and the GLP group, which are specialised for glycerol transport. The present study reports the identification and characterisation of a novel GLP family member in a teleost fish, the sea bream Sparus auratus. A sea bream aquaporin (sbAQP) cDNA of 1047·bp and encoding a protein of 298·amino acids was isolated from a kidney cDNA library. Functional characterization of the sbAQP using a Xenopus oocyte assay revealed that the isolated cDNA stimulated osmotic water permeability in a mercury-sensitive manner and also stimulated urea and glycerol uptake. Northern blotting demonstrated that sbAQP was expressed at high levels in the posterior region of the gut, where two transcripts were identified (1.6·kb and 2·kb), and in kidney, where a single transcript was present (2·kb). In situ hybridisation studies with a sbAQP riboprobe revealed its presence in the lamina propria and smooth muscle layer of the posterior region of the gut and in epithelial cells of some kidney tubules. sbAQP was also present in putative chloride cells of the gill. Phylogenetic analysis of sbAQP, including putative GLP genes from Fugu rubripes, revealed that it did not group with any of the previously isolated vertebrate GLPs and instead formed a separate group, suggesting that it may be a novel GLP member.
- Marine fish egg hydration is aquaporin-mediatedPublication . Fabra, Mercedes; Raldúa, Demetrio; Power, Deborah; Deen, P. M. T.; Cerda, J.The oocytes of marine pelagic teleosts swell abruptly by hydration during the reinitiation of meiosis (oocyte maturation) before ovulation. This process renders the eggs and early embryos buoyant in sea water, allowing their survival and dispersal in the ocean (1, 2). The major osmotic effectors for water uptake are free amino acids derived from the proteolysis of the oocyte major yolk proteins (3), but the membrane-associated mechanisms involved remain unknown.
- 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.
- Olfactory sensitivity to bile fluid and bile salts in the European eel (Anguilla anguilla), goldfish (Carassius auratus) and Mozambique tilapia (Oreochromis mossambicus) suggests a 'broad range' sensitivity not confined to those produced by conspecifics alonePublication . Huertas, Mar; Hagey, L.; Hofmann, A. F.; Cerda, J.; Canario, Adelino V. M.; Hubbard, PeterTeleosts have high olfactory sensitivity to bile salts. To assess whether this phenomenon is involved in intra-specific chemical communication alone, or is part of a more ‘broad range’ sensitivity to bile salts produced by heterospecifics, we investigated possible differences in the odour of bile between the sexes and among different species – the eel (Anguilla anguilla), goldfish (Carassius auratus) and Mozambique tilapia (Oreochromis mossambicus) – using the electro-olfactogram (EOG). We also identified the main bile constituents by liquid chromatography and mass spectrometry. There were marked differences in olfactory response of the eel to thin-layer chromatography fractions of bile from both sexes, and mature and immature conspecifics. Smaller differences were seen in the potency of fractions of bile from male and female goldfish and tilapia. Eels, goldfish and tilapia demonstrated similar olfactory sensitivity to bile from a range of different species, with no apparent correlation between the olfactory potency of bile and a phylogenetic closeness and/or similarity of diet of the donor to the receiver. The three species were able to detect odorants in thin-layer chromatography fractions of heterospecific bile even in the absence of activity in conspecific bile. Eels, goldfish and tilapia responded to both sulphated C27 bile salts (5-scymnol-sulphate and 5-cyprinol sulphate) and to taurine-conjugated C24 bile salts (taurochenodeoxycholic acid, taurolithocholic acid and taurocholic acid), irrespective of whether these bile salts were present in conspecific bile. Together, these results suggest that teleosts have a broad-range olfactory sensitivity to bile salts, with potential roles in both intra-specific chemical communication and in inter-specific interactions.
- Olfactory sensitivity to conspecific bile fluid and skin mucus in the European eel Anguilla anguilla (L.)Publication . Huertas, Mar; Hubbard, Peter; Canario, Adelino V. M.; Cerda, J.The present study assessed the olfactory potency of conspecific bile fluid and skin mucus in the European eel Anguilla anguilla by the electro-olfactogram. Immature males showed high olfactory sensitivity to conspecific bile, giving large amplitude responses in a concentrationdependent manner with estimated thresholds of detection of <1:107 (n = 6). Mucus also proved to contain highly potent odorants with thresholds of detection of c. 1:106 (n = 6). Crude solidphase extraction of bile fluid (C-18 and C-2/ENV+ cartridges) showed that the majority of olfactory activity in bile fluid was contained in the eluate of C-18 cartridges (n = 6). There were quantitative differences, however, between the sexes; female bile fluid had a higher proportion of activity in this fraction. Similar solid-phase extraction of mucus showed that it contains a higher proportion of odorants in the C-18 filtrate than bile fluid. Mucus from mature eels, however, had a higher proportion of olfactory activity in the eluate than immature fish (n = 6). Cross-adaptation experiments suggest that there are qualitative differences in the odorants contained in bile and mucus depending on both the sex and state of sexual maturation of the donor (n = 6). These results are consistent with a role for chemical communication in the reproduction of the European eel and suggest that both bile and mucus are potential sources of the odorants involved.
- Sexually mature European eels (Anguilla anguilla L.) stimulate gonadal development of neighbouring males: possible involvement of chemical communicationPublication . Huertas, Mar; Scott, A. P.; Hubbard, Peter; Canario, Adelino V. M.; Cerda, J.This study was aimed to investigate whether sexual maturation of immature male eels could be stimulated indirectly by placing them in contact with either male (Minj) or female (Finj) eels in which sexual maturation had been stimulated directly by weekly injections of human chorionic gonadotropin (hCG) or salmon pituitary extract (SPE), respectively. Untreated males were placed either in the same tank or in a separate tank that was linked to the injected Wsh via a recirculation system. The hormonal treatments stimulated spermatogenesis and spermiation in Minj, and ovulation in Finj as well as an increase of the ocular (Io) and gonadosomatic (GSI) indices in both sexes. Plasma levels of testosterone (T) and 11-ketotestosterone (11-KT) increased in Minj and T and 17 -estradiol (E2) in Finj. A small peak of plasma 17,20 -dihydroxypregn-4-en-3-one (17,20 P) occurred during ovulation, while the plasma levels of 17 -hydroxypregn-4- ene-3,20-dione (17P) were undetectable in both males and females. The water conditioned by Minj and Finj induced signiWcant, though relatively minor, increases in Io and GSI in uninjected males. In addition, uninjected Wsh showed small changes in plasma T and 11-KT levels, apparently related to the timing of spermiation and ovulation of Minj and Finj, respectively, as well as an activation of spermatogenesis (but not spermiation). Injected Wsh released free and conjugated T, 11-KT and E2 into the water, although immature eels were unable to smell (by electro-olfactogram) any of these steroids or prostaglandin F2 . However, immature males were highly sensitive to water extracts conditioned by spermiating Minj and pre-ovulatory and ovulated Finj. These preliminary results suggest the existence of chemical communication between maturing eels and immature males that stimulates gonad development, although the putative pheromone(s) involved has/have not yet been identiWed.