61 research outputs found
Role of estrogens in fish immunity with special emphasis on GPER1
It is well accepted that estrogens, the primary female sex hormones, play a key role in modulating different aspects of the immune response. Moreover, estrogens have been linked with the sexual dimorphism observed in some immune disorders, such as chronic inflammatory and autoimmune diseases. Nevertheless, their effects are often controversial and depend on several factors, such as the pool of estrogen receptors (ERs) involved in the response. Their classical mode of action is through nuclear ERs, which act as transcription factors, promoting the regulation of target genes. However, it has long been noted that some of the estrogen-mediated effects cannot be explained by these classical receptors, since they are rapid and mediated by non-genomic signaling pathways. Hence, the interest in membrane ERs, especially in G protein-coupled estrogen receptor 1 (GPER1), has grown in recent years. Although the presence of nuclear ERs, and ER signaling, in immune cells in mammals and fish has been well documented, information on membrane ERs is much scarcer. In this context, the present manuscript aims to review our knowledge concerning the effect of estrogens on fish immunity, with special emphasis on GPER1. For example, the numerous tools developed over recent years allowed us to report for the first time that the regulation of fish granulocyte functions by estrogens through GPER1 predates the split of fish and tetrapods more than 450 million years ago, pointing to the relevance of estrogens as modulators of the immune responses, and the pivotal role of GPER1 in immunity.Versión del editor3,26
Tamoxifen disrupts the reproductive process in gilthead seabream males and modulates the effects promoted by 17alpha-ethynylestradiol
17α-ethynylestradiol (EE2), which is used in oral contraceptives and hormone replacement therapy, is a well documented estrogenic endocrine disruptor and an aquatic contaminant. In the present study, adult male specimens of the marine hermaphrodite teleost gilthead (Sparus aurata L.) were fed a diet containing tamoxifen (Tmx), an estrogen receptor ligand used in cancer therapy, alone or combined with EE2, for 25 days and then fed a commercial diet for a further 25 days (recovery period). The effects of short (5 days) and long (25 days) treatments on several reproductive and gonad immune parameters and the reversibility of the disruptive effects after the recovery period were examined. Our data showed that Tmx acted as an estrogenic endocrine disruptor as revealed by the increase in the hepatic transcription of the vitellogenin gene in males, the serum levels of 17β-estradiol and the gonad expression levels of the estrogen receptor α and G protein-coupled estrogen receptor genes, and the recruitment of leukocytes into the gonad, a well known estrogenic-dependent process in gilthead seabream males. On the other hand, Tmx also increased sperm concentration and motility as well as the serum levels of androgens and the expression levels of genes that codify for androgenic enzymes, while decreasing the expression levels of the gene that code for gonadal aromatase. When applied simultaneously, Tmx and EE2 could act in synergy or counteract, each other, depending on the parameter measured. The disruptive effect of EE2 and/or Tmx was not reversible after a 25 day recovery period.Postprin
Vaccination of Gilthead Seabream After Continuous Xenoestrogen Oral Exposure Enhances the Gut Endobolome and Immune Status via GPER1
In fish culture settings, the exogenous input of steroids is a matter of concern. Recently, we unveiled that in the gilthead seabream (Sparus aurata), the G protein-coupled estrogen receptor agonist G-1 (G1) and the endocrine disruptor 17α-ethinylestradiol (EE2) are potent modulators in polyreactive antibody production. However, the integral role of the microbiota upon immunity and antibody processing in response to the effect of EE2 remains largely unexplored. Here, juvenile seabreams continuously exposed for 84 days to oral G1 or EE2 mixed in the fish food were intraperitoneally (i.p.) immune primed on day 42 with the model antigen keyhole limpet hemocyanin (KLH). A critical panel of systemic and mucosal immune markers, serum VTG, and humoral, enzymatic, and bacteriolytic activities were recorded and correlated with gut bacterial metagenomic analysis 1 day post-priming (dpp). Besides, at 15 dpp, animals received a boost to investigate the possible generation of specific anti-KLH antibodies at the systemic and mucosal interphases by the end of the trial. On day 43, EE2 but not G1 induced a significant shift in the serum VTG level of naive fish. Simultaneously, significant changes in some immune enzymatic activities in the serum and gut mucus of the EE2-treated group were recorded. In comparison, the vaccine priming immunization resulted in an attenuated profile of most enzymatic activities in the same group. The gut genes qPCR analysis exhibited a related pattern, only emphasized by a significant shift in the EE2 group’s il1b expression. The gut bacterial microbiome status underwent 16S rRNA dynamic changes in alpha diversity indices, only with the exposure to oral G1, supporting functional alterations on cellular processes, signaling, and lipid metabolism in the microbiota. By the same token, the immunization elevated the relative abundance of Fusobacteria only in the control group, while this phylum was depleted in both the treated groups. Remarkably, the immunization also promoted changes in the bacterial class Betaproteobacteria and the estrogen-associated genus Novosphingobium. Furthermore, systemic and mucosal KLH-specific immunoglobulin (Ig)M and IgT levels in the fully vaccinated fish showed only slight changes 84 days post-estrogenic oral administration. In summary, our results highlight the intrinsic relationship among estrogens, their associated receptors, and immunization in the ubiquitous fish immune regulation and the subtle but significant crosstalk with the gut endobolome.Versión del edito
17α‑ethynylestradiol prevents the natural male‑to‑female sex change in gilthead seabream (Sparus aurata L.)
Exposure to 17α-ethynylestradiol (EE2, 5 μg/g food) impairs some reproductive events in the protandrous gilthead seabream and a short recovery period does not allow full recovery. In this study, spermiating seabream males in the second reproductive cycle (RC) were fed a diet containing 5 or 2.5 μg EE2/g food for 28 days and then a commercial diet without EE2 for the remaining RC. Individuals were sampled at the end of the EE2 treatment and then at the end of the RC and at the beginning of the third RC, 146 and 333 days after the cessation of treatment, respectively. Increased hepatic transcript levels of the gene coding for vitellogenin (vtg) and plasma levels of Vtg indicated both concentrations of EE2 caused endocrine disruption. Modifications in the histological organization of the testis, germ cell proliferation, plasma levels of the sex steroids and pituitary expression levels of the genes coding for the gonadotropin β-subunits, fshβ and lhβ were detected. The plasma levels of Vtg and most of the reproductive parameters were restored 146 days after treatments. However, although 50% of the control fish underwent sex reversal as expected at the third RC, male-to female sex change was prevented by both EE2
concentrations.Ministerio de Ciencia e Innovación and FEDER (AGL2014-53167-C3-1R, -2-R; AGL2017-85978-C2-1-R; RTI2018-096625-B-C33) and the Fundación Séneca (CARM) (19883/GERM/15).Versión del editor2,92
Endocrine disrupter chemicals affect the humoral antimicrobial activities of gilthead seabream males even upon the cease of the exposure
17α-ethynilestradiol (EE2) and tamoxifen (Tmx) are pollutants world-wide distributed in aquatic environments. Gilthead seabream, Sparus aurata L., is highlighted as a species model of intensively culture in anthropogenic disturbed environments. The effects of these pollutants on gilthead seabream reproduction and some immune responses have been described but, the humoral innate antimicrobial activities have never received attention. In this work we analysed the latest in the plasma of gilthead seabream males of different ages and reproductive stages treated with 0, 2.5, 5 or 50 μg EE2 or 100 μg Tmx g−1 food during different times of exposure and of reverting to commercial diet (recovery). The peroxidase and protease activities decreased as the spermatogenesis of the first reproductive cycle (RC) proceeded in control fish. However, only protease and antiprotease activities showed different level at different stages of the second RC in control fish, but showed scarce disruption in fish treated with EE2 or Tmx. Peroxidase and bactericide activities are more sensitive to EE2, than to Tmx. The effects induced by EE2 varied depending on the activity analyzed, the dose and the time of exposure and the reproductive stage and the age of the specimens.Versión del editor2,92
Nanoencapsulated Clove Oil Applied as an Anesthetic at Slaughtering Decreases Stress, Extends the Freshness, and Lengthens Shelf Life of Cultured Fish
In the aquaculture industry, fish are stunned using a wide range of methods, but all of them trigger stress responses and affect the fish flesh quality. Chilled water is considered one of the most efficient methods, but even this is not a stress‐free experience for the fish. Anesthetics included in the ice slurry or in water could decrease this stress and delay the loss of flesh quality. In this work, we analyze the effect of clove oil (CO) nanoencapsulated in β‐cyclodextrins (β‐CD) (CO + β‐CD), incorporated in the stunning bath, on the stress response and the organoleptic attributes of fresh marine and freshwater fish from four economically important fish species: Atlantic salmon, European seabass, Nile tilapia, and Rainbow trout. CO + β‐CD reduces the time required to induce anesthesia, independently of water salinity, habitat or water temperature. The plasmatic glucose and cortisol levels decreased in all four species, although the concentrations of CO varied between species. Moreover, plasmatic lactate level differed between the marine and freshwater fish. The use of CO + β‐CD extended the shelf life of fish from all the species studied (by 3–7 days). In conclusion, using CO encapsulated in β‐CD for anesthetizing fish can be regarded as an improved fish‐stunning technique that reduces the anesthesia‐induction time, decreases the stress response, and extends the shelf life of fresh fish.Versión del edito
The Effect of 17α-Ethynilestradiol and GPER1 Activation on Body and Muscle Growth, Muscle Composition and Growth-Related Gene Expression of Gilthead Seabream, Sparus aurata L
Endocrine-disrupting chemicals include natural and synthetic estrogens, such as 17α- ethynilestradiol (EE2
), which can affect reproduction, growth and immunity. Estrogen signalling is
mediated by nuclear or membrane estrogen receptors, such as the new G-protein-coupled estrogen receptor 1 (GPER1). The present work studies the effect of EE2
and G1 (an agonist of GPER1) on
body and muscle parameters and growth-related genes of 54 two-year-old seabreams. The fish were fed a diet containing EE2
(EE2 group) and G1 (G1 group) for 45 days and then a diet without EE2 or
G1 for 122 days. An untreated control group was also studied. At 45 days, the shortest body length was observed in the G1 group, while 79 and 122 days after the cessation of treatments, the shortest body growth was observed in the EE2
group. Hypertrophy of white fibers was higher in the EE2 and
G1 groups than it was in the control group, whereas the opposite was the case with respect to hy- perplasia. Textural hardness showed a negative correlation with the size of white fibers. At the end of the experiment, all fish analyzed in the EE2
group showed a predominance of the gonadal ovarian
area. In addition, the highest expression of the mafbx gene (upregulated in catabolic signals) and mstn2 (myogenesis negative regulator) was found in EE2
-exposed fish.Versión del editor2,46
Nodavirus colonizes and replicates in the testis of gilthead seabream and European sea bass modulating its immune and reproductive functions
Viruses are threatening pathogens for fish aquaculture. Some of them are transmitted
through gonad fluids or gametes as occurs with nervous necrosis virus (NNV). In order to
be transmitted through the gonad, the virus should colonize and replicate inside some cell
types of this tissue and avoid the subsequent immune response locally. However, whether
NNV colonizes the gonad, the cell types that are infected, and how the immune response in
the gonad is regulated has never been studied. We have demonstrated for the first time the
presence and localization of NNV into the testis after an experimental infection in the European
sea bass (Dicentrarchus labrax), and in the gilthead seabream (Sparus aurata), a very
susceptible and an asymptomatic host fish species, respectively. Thus, we localized in the
testis viral RNA in both species using in situ PCR and viral proteins in gilthead seabream by
immunohistochemistry, suggesting that males might also transmit the virus. In addition, we
were able to isolate infective particles from the testis of both species demonstrating that
NNV colonizes and replicates into the testis of both species. Blood contamination of the tissues
sampled was discarded by completely fish bleeding, furthermore the in situ PCR and
immunocytochemistry techniques never showed staining in blood vessels or cells. Moreover,
we also determined how the immune and reproductive functions are affected comparing
the effects in the testis with those found in the brain, the main target tissue of the virus.
Interestingly, NNV triggered the immune response in the European sea bass but not in the
gilthead seabream testis. Regarding reproductive functions, NNV infection alters 17β-estradiol
and 11-ketotestosterone production and the potential sensitivity of brain and testis to
these hormones, whereas there is no disruption of testicular functions according to several
reproductive parameters. Moreover, we have also studied the NNV infection of the testis in
vitro to assess local responses. Our in vitro results show that the changes observed on the expression of immune and reproductive genes in the testis of both species are different to
those observed upon in vivo infections in most of the casesMINECO and FEDER (AGL2010-20801-C02-01; AGL2010-20801-C02-02; AGL2013-43588-P); Fundación Séneca (04538/GERM/06)Versión del editor4,411
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