165 results on '"Peñaranda, D.S."'
Search Results
2. Rabbit morula vitrification reduces early foetal growth and increases losses throughout gestation
- Author
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Vicente, J.S., Saenz-de-Juano, M.D., Jiménez-Trigos, E., Viudes-de-Castro, M.P., Peñaranda, D.S., and Marco-Jiménez, F.
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- 2013
- Full Text
- View/download PDF
3. Standardization of European eel (Anguilla anguilla) sperm motility evaluation by CASA software
- Author
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Gallego, V., Carneiro, P.C.F., Mazzeo, I., Vílchez, M.C., Peñaranda, D.S., Soler, C., Pérez, L., and Asturiano, J.F.
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- 2013
- Full Text
- View/download PDF
4. Comparison of two techniques for the morphometry study on gilthead seabream (Sparus aurata) spermatozoa and evaluation of changes induced by cryopreservation
- Author
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Gallego, V., Peñaranda, D.S., Marco-Jiménez, F., Mazzeo, I., Pérez, L., and Asturiano, J.F.
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- 2012
- Full Text
- View/download PDF
5. Successful inclusion of high vegetable protein sources in feed for rainbow trout without decrement in intestinal health
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Vélez-Calabria, Glenda, Peñaranda, D.S., Jover Cerda, Miguel, Martínez-Llorens, Silvia, and Tomas-Vidal, A.
- Subjects
General Veterinary ,Fishmeal substitution ,Veterinary medicine ,Interleukins ,food and beverages ,PRODUCCION ANIMAL ,Article ,intestinal track ,fishmeal substitution ,amino acid supplement ,interleukins ,immune system ,inflammation response ,Immune system ,QL1-991 ,SF600-1100 ,Amino acid supplement ,Animal Science and Zoology ,Intestinal track ,Inflammation response ,Zoology - Abstract
Simple Summary A reduction in fishmeal in diets is essential to achieve the aim of sustainable production. In the current work, using a plant protein blend of wheat gluten, wheat and soybean meal supplemented with Tau, Val, Lys and Met, a 10% higher fishmeal substitution without affecting growth and health parameters has been accomplished. Abstract The aquaculture of carnivorous fish is in continuous expansion, which leads to the need to reduce the dependence on fishmeal (FM). Plant proteins (PP) represent a suitable protein alternative to FM and are increasingly used in fish feed. However, PP may lead to stunted growth and enteritis. In the current study, the effect of high FM substitution by PP sources on the growth, mortality and intestinal health of rainbow trout (Oncorhynchus mykiss) was evaluated in terms of the histological intestine parameters and expression of genes related to inflammation (IL-1β, IL-8 and TGF-β) and immune responses (Transferrin, IgT and IFN-γ). The results show that a total substitution registered lower growth and survival rates, probably due to a disruption to the animal’s health. Confirming this hypothesis, fish fed FM0 showed histological changes in the intestine and gene changes related to inflammatory responses, which in the long-term could have triggered an immunosuppression. The FM10 diet presented not only a similar expression to FM20 (control diet), but also similar growth and survival. Therefore, 90% of FM substitution was demonstrated as being feasible in this species using a PP blend of wheat gluten (WG) and soybean meal (SBM) as a protein source.
- Published
- 2021
6. Supplementary Figure S1
- Author
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Peñaranda, D.S.
- Abstract
Gene expression of IgT, Transferrin (Transf), IFN-γ, IL-8, IL-8, IL-1β and TGF-β in anterior and posterior intestine section
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- 2021
- Full Text
- View/download PDF
7. Improvement of European eel sperm cryopreservation method by preventing spermatozoa movement activation caused by cryoprotectants
- Author
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Peñaranda, D.S., Pérez, L., Gallego, V., Jover, M., and Asturiano, J.F.
- Published
- 2009
- Full Text
- View/download PDF
8. Estimation of phosphorous and nitrogen waste in rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) diets including different inorganic phosphorous sources
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, GLOBAL FEED, S.L.U., European Regional Development Fund, Centro para el Desarrollo Tecnológico Industrial, Milián-Sorribes, María Consolación, Tomas-Vidal, A., Peñaranda, D.S., Carpintero, Laura, Mesa, Juan S., Dupuy, Javier, Donadeu, Andrés, Macias-Vidal, Judith, Martínez-Llorens, Silvia, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, GLOBAL FEED, S.L.U., European Regional Development Fund, Centro para el Desarrollo Tecnológico Industrial, Milián-Sorribes, María Consolación, Tomas-Vidal, A., Peñaranda, D.S., Carpintero, Laura, Mesa, Juan S., Dupuy, Javier, Donadeu, Andrés, Macias-Vidal, Judith, and Martínez-Llorens, Silvia
- Abstract
[EN] Aquaculture effluents with high levels of phosphorus (P) and nitrogen (N) contribute to eutrophication in the aquatic ecosystem. The environmental impact of phosphorus and N aquaculture waste may be diminished by modifying diet ingredients that improve phosphorous (P) digestibility, and therefore, reduce the P in metabolic waste. The content of P in fishmeal is high (30 g/kg), but the inclusion of fishmeal in the diet is reducing due to its high costs and limited accessibility; therefore, the addition of an inorganic P source is necessary to ensure a satisfactory level of available P in fish diets. Consequently, the present study aimed to evaluate the effect of four different inorganic P sources on P digestibility and excretion in rainbow trout (Oncorhynchus mykiss), as one of the most relevant aquaculture species. Monosodium/monocalcium phosphate with 2% of sodium source presented a P digestibility similar to monoammonium phosphate, but with lower nitrogen and phosphorus excretion into the environment, which is advantageous from a nutritional, environmental and industrial point of view (biofilters and recirculation systems in fish farms). This study was conducted to evaluate the apparent availability and P and N excretion in rainbow trout (Oncorhynchus mykiss) using different inorganic phosphorus sources. With this goal, fish (153 +/- 14.1 g) fed four inorganic P sources were assayed: monoammonium phosphate (MAP, NH4H2PO4), monosodium/monocalcium phosphate (SCP-2%, AQphos+, NaH2PO4/Ca(H2PO4)(2)center dot H2O in proportion 12/88), monosodium/monocalcium phosphate (SCP-5%, NaH2PO4/Ca(H2PO4)(2)center dot H2O in proportion 30/70) and monocalcium phosphate (MCP, Ca(H2PO4)(2)center dot H2O). Phosphorus (P) digestibility, in diets that included MAP and SCP-2% as inorganic phosphorus sources, were significantly higher than for SCP-5% and MCP sources. In relation to the P excretion pattern, independent of the diet, a peak at 6 h after feeding was registered, but at dif
- Published
- 2021
9. Successful inclusion of high vegetable protein sources in feed for rainbow trout without decrement in intestinal health
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Vélez-Calabria, Glenda, Peñaranda, D.S., Jover Cerda, Miguel, Martínez-Llorens, Silvia, Tomas-Vidal, A., Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Vélez-Calabria, Glenda, Peñaranda, D.S., Jover Cerda, Miguel, Martínez-Llorens, Silvia, and Tomas-Vidal, A.
- Abstract
[EN] A reduction in fishmeal in diets is essential to achieve the aim of sustainable production. In the current work, using a plant protein blend of wheat gluten, wheat and soybean meal supplemented with Tau, Val, Lys and Met, a 10% higher fishmeal substitution without affecting growth and health parameters has been accomplished. The aquaculture of carnivorous fish is in continuous expansion, which leads to the need to reduce the dependence on fishmeal (FM). Plant proteins (PP) represent a suitable protein alternative to FM and are increasingly used in fish feed. However, PP may lead to stunted growth and enteritis. In the current study, the effect of high FM substitution by PP sources on the growth, mortality and intestinal health of rainbow trout (Oncorhynchus mykiss) was evaluated in terms of the histological intestine parameters and expression of genes related to inflammation (IL-1 beta, IL-8 and TGF-beta) and immune responses (Transferrin, IgT and IFN-gamma). The results show that a total substitution registered lower growth and survival rates, probably due to a disruption to the animal's health. Confirming this hypothesis, fish fed FM0 showed histological changes in the intestine and gene changes related to inflammatory responses, which in the long-term could have triggered an immunosuppression. The FM10 diet presented not only a similar expression to FM20 (control diet), but also similar growth and survival. Therefore, 90% of FM substitution was demonstrated as being feasible in this species using a PP blend of wheat gluten (WG) and soybean meal (SBM) as a protein source.
- Published
- 2021
10. Effect of embryo vitrification on the steroid biosynthesis of liver tissue in rabbit offspring
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Economía y Competitividad, Marco-Jiménez, Francisco, Garcia-Dominguez, X, Domínguez-Martínez, Marta, Viudes-de-Castro, María Pilar, Diretto, Gianfranco, Peñaranda, D.S., Vicente Antón, José Salvador, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Economía y Competitividad, Marco-Jiménez, Francisco, Garcia-Dominguez, X, Domínguez-Martínez, Marta, Viudes-de-Castro, María Pilar, Diretto, Gianfranco, Peñaranda, D.S., and Vicente Antón, José Salvador
- Abstract
[EN] Preimplantation embryo manipulations during standard assisted reproductive technologies (ART) have significant repercussions on offspring. However, few studies to date have investigated the potential long-term outcomes associated with the vitrification procedure. Here, we performed an experiment to unravel the particular effects related to stress induced by embryo transfer and vitrification techniques on offspring phenotype from the foetal period through to prepuberal age, using a rabbit model. In addition, the focus was extended to the liver function at prepuberal age. We showed that, compared to naturally conceived animals (NC), offspring derived after embryo exposure to the transfer procedure (FT) or cryopreservation-transfer procedure (VT) exhibited variation in growth and body weight from foetal life to prepuberal age. Strikingly, we found a nonlinear relationship between FT and VT stressors, most of which were already present in the FT animals. Furthermore, we displayed evidence of variation in liver function at prepuberal age, most of which occurred in both FT and VT animals. The present major novel finding includes a significant alteration of the steroid biosynthesis profile. In summary, here we provide that embryonic manipulation during the vitrification process is linked with embryo phenotypic adaptation detected from foetal life to prepuberal age and suggests that this phenotypic variation may be associated, to a great extent, with the effect of embryo transfer.
- Published
- 2020
11. Long-Term Phenotypic and Proteomic Changes Following Vitrified Embryo Transfer in the Rabbit Model
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Agencia Estatal de Investigación, Ministerio de Economía y Empresa, Garcia-Dominguez, X, Marco-Jiménez, Francisco, Peñaranda, D.S., Vicente Antón, José Salvador, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Agencia Estatal de Investigación, Ministerio de Economía y Empresa, Garcia-Dominguez, X, Marco-Jiménez, Francisco, Peñaranda, D.S., and Vicente Antón, José Salvador
- Abstract
[EN] This study was conducted to demonstrate how a vitrified embryo transfer procedure incurs phenotypic and molecular changes throughout life. This study reports the first evidence describing that embryonic manipulation during a vitrified embryo transfer cycle induced molecular modifications, concerning oxidative phosphorylation and dysregulations in zinc and lipid metabolism in liver tissue, which has been reported as responsible for postnatal variations of the phenotype. Nowadays, assisted reproductive technologies (ARTs) are considered valuable contributors to our past, but a future without their use is inconceivable. However, in recent years, several studies have evidenced a potential impact of ART on long-term development in mammal species. To date, the long-term follow-up data are still limited. So far, studies have mainly focused on in vitro fertilization or in vitro culture, with information from gametes/embryos cryopreservation field being practically missing. Herein, we report an approach to determine whether a vitrified embryo transfer procedure would have long-term consequences on the offspring. Using the rabbit as a model, we compared animals derived from vitrified-transferred embryos versus those naturally conceived, studying the growth performance, plus the weight throughout life, and the internal organs/tissues phenotype. The healthy status was assessed over the hematological and biochemical parameters in peripheral blood. Additionally, a comparative proteomic analysis was conducted in the liver tissue to investigate molecular cues related to vitrified embryo transfer in an adult tissue. After vitrified embryo transfer, birth weight was increased, and the growth performance was diminished in a sex-specific manner. In addition, vitrified-transferred animals showed significantly lower body, liver and heart weights in adulthood. Molecular analyses revealed that vitrified embryo transfer triggers reprogramming of the liver proteome. Functional analysis
- Published
- 2020
12. Impact of high dietary plant protein with or without marine ingredients in gut mucosa proteome of gilthead seabream (Sparus aurata, L.)
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Educación, Cultura y Deporte, Estruch, Guillem, Martínez-Llorens, Silvia, Tomas-Vidal, A., Monge-Ortiz, Raquel, Jover Cerda, Miguel, Brown, Paul B., Peñaranda, D.S., Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Educación, Cultura y Deporte, Estruch, Guillem, Martínez-Llorens, Silvia, Tomas-Vidal, A., Monge-Ortiz, Raquel, Jover Cerda, Miguel, Brown, Paul B., and Peñaranda, D.S.
- Abstract
[EN] The digestive tract, particularly the intestine, represents one of the main sites of interactions with the environment, playing the gut mucosa a crucial role in the digestion and absorption of nutrients, and in the immune defence. Previous researches have proven that the fishmeal replacement by plant sources could have an impact on the intestinal status at both digestive and immune level, compromising relevant productive parameters, such as feed efficiency, growth or survival. In order to evaluate the long-term impact of total fishmeal replacement on intestinal mucosa, the gut mucosa proteome was analysed in fish fed with a fishmeal-based diet, against plant protein-based diets with or without alternative marine sources inclusion. Total fishmeal replacement without marine ingredients inclusion, reported a negative impact in growth and biometric parameters, further an altered gut mucosa proteome. However, the inclusion of a low percentage of marine ingredients in plant protein-based diets was able to maintain the growth, biometrics parameters and gut mucosa proteome with similar values to FM group. A total fishmeal replacement induced a big set of underrepresented proteins in relation to several biological processes such as intracellular transport, assembly of cellular macrocomplex, protein localization and protein catabolism, as well as several molecular functions, mainly related with binding to different molecules and the maintenance of the cytoskeleton structure. The set of downregulated proteins also included molecules which have a crucial role in the maintenance of the normal function of the enterocytes, and therefore, of the epithelium, including permeability, immune and inflammatory response regulation and nutritional absorption. Possibly, the amino acid imbalance presented in VM diet, in a long-term feeding, may be the main reason of these alterations, which can be prevented by the inclusion of 15% of alternative marine sources. Significance: Long-term f
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- 2020
13. Intestinal Explant Cultures from Gilthead Seabream (Sparus aurata, L.) Allowed the Determination of Mucosal Sensitivity to Bacterial Pathogens and the Impact of a Plant Protein Diet
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Educación, Ministerio de Economía y Competitividad, Peñaranda, D.S., Bäuerl, Christine, Tomas-Vidal, A., Jover Cerda, Miguel, Estruch, Guillem, Pérez Martínez, Gaspar, Martínez-Llorens, Silvia, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Educación, Ministerio de Economía y Competitividad, Peñaranda, D.S., Bäuerl, Christine, Tomas-Vidal, A., Jover Cerda, Miguel, Estruch, Guillem, Pérez Martínez, Gaspar, and Martínez-Llorens, Silvia
- Abstract
[EN] The interaction between diet and intestinal health has been widely discussed, although in vivo approaches have reported limitations. The intestine explant culture system developed provides an advantage since it reduces the number of experimental fish and increases the time of incubation compared to similar methods, becoming a valuable tool in the study of the interactions between pathogenic bacteria, rearing conditions, or dietary components and fish gut immune response. The objective of this study was to determine the influence of the total substitution of fish meal by plants on the immune intestinal status of seabream using an ex vivo bacterial challenge. For this aim, two growth stages of fish were assayed (12 g): phase I (90 days), up to 68 g, and phase II (305 days), up to 250 g. Additionally, in phase II, the effects of long term and short term exposure (15 days) to a plant protein (PP) diet were determined. PP diet altered the mucosal immune homeostasis, the younger fish being more sensitive, and the intestine from fish fed short-term plant diets showed a higher immune response than with long-term feeding. Vibrio alginolyticus (V. alginolyticus) triggered the highest immune and inflammatory response, while COX-2 expression was significantly induced by Photobacterium damselae subsp. Piscicida (P. damselae subsp. Piscicida), showing a positive high correlation between the pro-inflammatory genes encoding interleukin 1 beta (IL1-beta), interleukin 6 (IL-6) and cyclooxygenase 2(COX-2).
- Published
- 2020
14. Long-term and transgenerational phenotypic, transcriptional and metabolic effects in rabbit males born following vitrified embryo transfer
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, GENERALITAT VALENCIANA, MINISTERIO DE ECONOMIA Y EMPRESA, Garcia-Dominguez, X, Marco-Jiménez, Francisco, Peñaranda, D.S., Diretto, Gianfranco, García-Carpintero, Víctor, Cañizares Sales, Joaquín, Vicente Antón, José Salvador, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, GENERALITAT VALENCIANA, MINISTERIO DE ECONOMIA Y EMPRESA, Garcia-Dominguez, X, Marco-Jiménez, Francisco, Peñaranda, D.S., Diretto, Gianfranco, García-Carpintero, Víctor, Cañizares Sales, Joaquín, and Vicente Antón, José Salvador
- Abstract
[EN] The advent of assisted reproductive technologies (ART) in mammals involved an extraordinary change in the environment where the beginning of a new organism takes place. Under in vitro conditions, in which ART is currently being performed, it likely fails to mimic optimal in vivo conditions. This suboptimal environment could mediate in the natural developmental trajectory of the embryo, inducing lasting effects until later life stages that may be inherited by subsequent generations (transgenerational effects). Therefore, we evaluated the potential transgenerational effects of embryo exposure to the cryopreservation-transfer procedure in a rabbit model on the offspring phenotype, molecular physiology of the liver (transcriptome and metabolome) and reproductive performance during three generations (F1, F2 and F3). The results showed that, compared to naturally-conceived animals (NC group), progeny generated after embryo exposure to the cryopreservation-transfer procedure (VT group) exhibited lower body growth, which incurred lower adult body weight in the F1 (direct effects), F2 (intergenerational effects) and F3 (transgenerational effects) generations. Furthermore, VT animals showed intergenerational effects on heart weight and transgenerational effects on liver weight. The RNA-seq data of liver tissue revealed 642 differentially expressed transcripts (DETs) in VT animals from the F1 generation. Of those, 133 were inherited from the F2 and 120 from the F3 generation. Accordingly, 151, 190 and 159 differentially accumulated metabolites (DAMs) were detected from the F1, F2 and F3, respectively. Moreover, targeted metabolomics analysis demonstrated that transgenerational effects were mostly presented in the non-polar fraction. Functional analysis of molecular data suggests weakened zinc and fatty acid metabolism across the generations, associated with alterations in a complex molecular network affecting global hepatic metabolism that could be associated with the phe
- Published
- 2020
15. Effects of hCG as spermiation inducer on European eel semen quality
- Author
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Asturiano, J.F., Marco-Jiménez, F., Pérez, L., Balasch, S., Garzón, D.L., Peñaranda, D.S., Vicente, J.S., Viudes-de-Castro, M.P., and Jover, M.
- Published
- 2006
- Full Text
- View/download PDF
16. Morphometry characterisation of European eel spermatozoa with computer-assisted spermatozoa analysis and scanning electron microscopy
- Author
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Marco-Jiménez, F., Pérez, L., Castro, M.P. Viudes de, Garzón, D.L., Peñaranda, D.S., Vicente, J.S., Jover, M., and Asturiano, J.F.
- Published
- 2006
- Full Text
- View/download PDF
17. Cryopreservation of European eel ( Anguilla anguilla) spermatozoa: Effect of dilution ratio, foetal bovine serum supplementation, and cryoprotectants
- Author
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Marco-Jiménez, F., Garzón, D.L., Peñaranda, D.S., Pérez, L., Viudes-de-Castro, M.P., Vicente, J.S., Jover, M., and Asturiano, J.F.
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- 2006
- Full Text
- View/download PDF
18. Cold seawater induces early maturational stages in the BPG axis of European eel males
- Author
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Rozenfeld, Christoffer, García-Carpintero-Burgos, Víctor, Pérez Igualada, Luz María, Gallego Albiach, Victor, Herranz-Jusdado, Juan Germán, Tveiten, Helge, Johnsen, Helge K., Fontaine, Romain, Weltzien, F.A., Cañizares Sales, Joaquín, Asturiano, Juan F., and Peñaranda, D.S.
- Subjects
GENETICA ,Histology ,Anguilla anguilla ,Immunofluorescence ,RNA-sequencing ,Temperature ,Spermatogonial proliferation ,Radioimmunoassay ,Epigenetics ,PRODUCCION ANIMAL ,Migration - Abstract
[EN] BackgroundThe impossibility of closing the life cycle of the European eel (Anguilla anguilla) in captivity troubles the future of this critically endangered species. In addition, the European eel is a highly valued and demanded resource, thus the successful closing of its life cycle would have a substantial economic and ecological impact. With the aim of obtaining the highest gamete quality, the study of the effects of environmental factors, such as temperature, on reproductive performance may prove valuable. This is especially true for the exposure to cold water, which has been reported to improve sexual development in multiple other Actinopterygii species.ResultsEuropean eel males treated with cold seawater (10 degrees C, T10) for 2weeks showed an increase in the proliferation and differentiation of spermatogonial cells until the differentiated spermatogonial type A cell stage, and elevated testosterone and 11-ketotestosterone plasma levels. Transcriptomes from the tissues of the brain-pituitary-gonad (BPG) axis of T10 samples revealed a differential gene expression profile compared to the other experimental groups, with clustering in a principal component analysis and in heat maps of all differentially expressed genes. Furthermore, a functional analysis of differentially expressed genes revealed enriched gene ontology terms involved in the regulation of circadian rhythm, histone modification, meiotic nuclear division, and others.ConclusionsCold seawater treatment had a clear effect on the activity of the BPG-axis of European eel males. In particular, our cold seawater treatment induces the synchronization and increased proliferation and differentiation of specific spermatogonial cells. In the transcriptomic results, genes related to thermoception were observed. This thermoception may have caused the observed effects through epigenetic mechanisms, since all analysed tissues further revealed differentially expressed genes involved in histone modification. The presented results support our hypothesis that a low temperature seawater treatment induces an early sexual developmental stage in European eels. This hypothesis is logical given that the average temperature experienced by eels in the early stages of their oceanic reproductive migration is highly similar to that of this cold seawater treatment. Further studies are needed to test whether a cold seawater treatment can improve the response of European eels to artificial hormonal treatment, as the results suggest., This work was subsidized by the Spanish Ministry of Science and Innovation (REPRO-TEMP project, AGL2013-41646-R), the Spanish Ministry of Innovation, Science and Universities (EELGONIA project, RTI2018-096413-B-I00), and by the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 642893 (IMPRESS), including the CR and JGHJ predoctoral contracts. JGHJ was granted a Short Term Scientific Mission by the COST Office (COST Action AQUAGAMETE: Assessing and improving the quality of aquatic animal gametes to enhance aquatic resources. The need to harmonize and standardize evolving methodologies, and improve transfer from academia to industry). The funding bodies had no role in the study, analysis, interpretation of data, or in writing the manuscript.
- Published
- 2019
19. Cold seawater induces early maturational stages in the BPG axis of European eel males
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, Agencia Estatal de Investigación, Ministerio de Economía y Empresa, European Commission, European Cooperation in Science and Technology, Rozenfeld, Christoffer, García-Carpintero-Burgos, Víctor, Pérez Igualada, Luz María, Gallego Albiach, Victor, Herranz-Jusdado, Juan Germán, Tveiten, Helge, Johnsen, Helge K., Fontaine, Romain, Weltzien, F.A., Cañizares Sales, Joaquín, Asturiano, Juan F., Peñaranda, D.S., Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, Agencia Estatal de Investigación, Ministerio de Economía y Empresa, European Commission, European Cooperation in Science and Technology, Rozenfeld, Christoffer, García-Carpintero-Burgos, Víctor, Pérez Igualada, Luz María, Gallego Albiach, Victor, Herranz-Jusdado, Juan Germán, Tveiten, Helge, Johnsen, Helge K., Fontaine, Romain, Weltzien, F.A., Cañizares Sales, Joaquín, Asturiano, Juan F., and Peñaranda, D.S.
- Abstract
[EN] BackgroundThe impossibility of closing the life cycle of the European eel (Anguilla anguilla) in captivity troubles the future of this critically endangered species. In addition, the European eel is a highly valued and demanded resource, thus the successful closing of its life cycle would have a substantial economic and ecological impact. With the aim of obtaining the highest gamete quality, the study of the effects of environmental factors, such as temperature, on reproductive performance may prove valuable. This is especially true for the exposure to cold water, which has been reported to improve sexual development in multiple other Actinopterygii species.ResultsEuropean eel males treated with cold seawater (10 degrees C, T10) for 2weeks showed an increase in the proliferation and differentiation of spermatogonial cells until the differentiated spermatogonial type A cell stage, and elevated testosterone and 11-ketotestosterone plasma levels. Transcriptomes from the tissues of the brain-pituitary-gonad (BPG) axis of T10 samples revealed a differential gene expression profile compared to the other experimental groups, with clustering in a principal component analysis and in heat maps of all differentially expressed genes. Furthermore, a functional analysis of differentially expressed genes revealed enriched gene ontology terms involved in the regulation of circadian rhythm, histone modification, meiotic nuclear division, and others.ConclusionsCold seawater treatment had a clear effect on the activity of the BPG-axis of European eel males. In particular, our cold seawater treatment induces the synchronization and increased proliferation and differentiation of specific spermatogonial cells. In the transcriptomic results, genes related to thermoception were observed. This thermoception may have caused the observed effects through epigenetic mechanisms, since all analysed tissues further revealed differentially expressed genes involved in histone modification. The
- Published
- 2019
20. De novo European eel transcriptome provides insights into the evolutionary history of duplicated genes in teleost lineages
- Author
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Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Economía y Empresa, European Commission, Rozenfeld, Christoffer, Blanca Postigo, José Miguel, Gallego Albiach, Victor, García-Carpintero, Víctor, Herranz-Jusdado, Juan Germán, Pérez Igualada, Luz María, Asturiano, Juan F., Cañizares Sales, Joaquín, Peñaranda, D.S., Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes, Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Economía y Empresa, European Commission, Rozenfeld, Christoffer, Blanca Postigo, José Miguel, Gallego Albiach, Victor, García-Carpintero, Víctor, Herranz-Jusdado, Juan Germán, Pérez Igualada, Luz María, Asturiano, Juan F., Cañizares Sales, Joaquín, and Peñaranda, D.S.
- Abstract
[EN] Paralogues pairs are more frequently observed in eels (Anguilla sp.) than in other teleosts. The paralogues often show low phylogenetic distances; however, they have been assigned to the third round of whole genome duplication (WGD), shared by all teleosts (3R), due to their conserved synteny. The apparent contradiction of low phylogenetic difference and 3R conserved synteny led us to study the duplicated gene complement of the freshwater eels. With this aim, we assembled de novo transcriptomes of two highly relevant freshwater eel species: The European (Anguilla anguilla) and the Japanese eel (Anguilla japonica). The duplicated gene complement was analysed in these transcriptomes, and in the genomes and transcriptomes of other Actinopterygii species. The study included an assessment of neutral genetic divergence (4dTv), synteny, and the phylogenetic origins and relationships of the duplicated gene complements. The analyses indicated a high accumulation of duplications (1217 paralogue pairs) among freshwater eel genes, which may have originated in a WGD event after the Elopomorpha lineage diverged from the remaining teleosts, and thus not at the 3R. However, very similar results were observed in the basal Osteoglossomorpha and Clupeocephala branches, indicating that the specific genomic regions of these paralogues may still have been under tetrasomic inheritance at the split of the teleost lineages. Therefore, two potential hypotheses may explain the results: i) The freshwater eel lineage experienced an additional WGD to 3R, and ii) Some duplicated genomic regions experienced lineage specific rediploidization after 3R in the ancestor to freshwater eels. The supporting/opposing evidence for both hypotheses is discussed.
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- 2019
21. Handling and treatment of male European eels (Anguilla anguilla) for hormonal maturation and sperm cryopreservation
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Herranz-Jusdado, Juan Germán, Kása, É, Kollár, Timea, Gallego Albiach, Victor, Peñaranda, D.S., Rozenfeld, Christoffer, Pérez Igualada, Luz María, Horváth, Ákos, and Asturiano Nemesio, Juan Francisco
- Subjects
Cryopreservation ,Male ,endocrine system ,Eels ,BIOLOGIA ANIMAL ,urogenital system ,Animals ,PRODUCCION ANIMAL ,Spermatozoa ,reproductive and urinary physiology ,Semen Preservation ,Developmental Biology - Abstract
[EN] During the last years, several research groups have been working on the development and improvement of new protocols for the European eel handling and maturation. As of yet, weekly injections of human chorionic gonadotropin (hCG) have proved to maturate males after just 5-6 weeks of treatment, producing high volumes of high-quality sperm during several weeks. In addition, sperm cryopreservation protocols using different extenders, cryoprotectants and cooling and thawing times have been previously described for European eel. Here, we show that Tanaka¿s extender solution can be directly used for fertilization or for cryopreservation, making unnecessary the usage of different types of solutions and dilutions. Furthermore, the use of methanol as a cryoprotectant makes this protocol easy to use as methanol has low toxicity and does not activate the sperm. The sperm does not need to be cryopreserved immediately after the addition of the cryoprotectant, and it can be used long after being thawed. Moreover, sperm motility is still high after thawing although it is lower than that of fresh sperm. The aim of this work is to show the best available protocol for European eel handling, maturation, and sperm cryopreservation., This publication was funded by the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 642893 (IMPRESS), the COST Office (COST Action FA 1205, AQUAGAMETE), and the Research Centre of Excellence -1476-4/2016/FEKUT
- Published
- 2018
22. Using specific recombinant gonadotropins to induce spermatogenesis and spermiation in the European eel (Anguilla anguilla)
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, European Commission, Ministerio de Economía, Industria y Competitividad, Peñaranda, D.S., Gallego Albiach, Victor, Rozenfeld, Christoffer, Herranz-Jusdado, Juan Germán, Pérez Igualada, Luz María, Gómez, A., Giménez, I., Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, European Commission, Ministerio de Economía, Industria y Competitividad, Peñaranda, D.S., Gallego Albiach, Victor, Rozenfeld, Christoffer, Herranz-Jusdado, Juan Germán, Pérez Igualada, Luz María, Gómez, A., Giménez, I., and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] New specific European eel (Anguilla anguilla) recombinant gonadotropins (aarGths) produced in the ovarian cells of Chinese hamsters (CHO) were used to induce maturation in captive male eels. In the first experiment, five different hormonal treatments were assayed: one group was given a constant dose of recombinant European eel follicle-stimulating hormone (aarFsh; 4 ¿g/fish) for 9 weeks, and the second group received a constant dose of recombinant European eel luteinizing hormone (aarLh; 2 ¿g/fish) also for 9 weeks. The other three groups were injected with different combinations of both aarGths (some doses constant, some variable). All five treatments stimulated androgen synthesis, but the increase was more pronounced in the fish treated with a combination of both aarGths. Unlike aarLh, aarFsh alone was able to induce spermiation, the best results were achieved in the fish that were treated with a constant dose of aarFSH and an increasing dose of aarLH, with spermiation being induced (20% motile cells) despite the fact that these fish were immature at the start of the experiment. In order to improve sperm quality, a second experiment was performed. Immature males received three constant doses of aarFsh (2.8, 1.4 or 0.7 ¿g/fish) and increasing doses of aarLh (every 3 weeks; 1, 2, 6 ¿g/fish). All the treatments induced spermiation, however the best sperm quality (with ¿50% motile cells) was observed in the males treated with the highest dose of aarFsh. In conclusion, these specific recombinant gonadotropins have demonstrated their capacity to induce spermatogenesis and spermiation in vivo in a teleost fish, the European eel.
- Published
- 2018
23. Handling and treatment of male European eels (Anguilla anguilla) for hormonal maturation and sperm cryopreservation
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, European Cooperation in Science and Technology, Herranz-Jusdado, Juan Germán, Kása, É, Kollár, Timea, Gallego Albiach, Victor, Peñaranda, D.S., Rozenfeld, Christoffer, Pérez Igualada, Luz María, Horváth, Ákos, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, European Cooperation in Science and Technology, Herranz-Jusdado, Juan Germán, Kása, É, Kollár, Timea, Gallego Albiach, Victor, Peñaranda, D.S., Rozenfeld, Christoffer, Pérez Igualada, Luz María, Horváth, Ákos, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] During the last years, several research groups have been working on the development and improvement of new protocols for the European eel handling and maturation. As of yet, weekly injections of human chorionic gonadotropin (hCG) have proved to maturate males after just 5-6 weeks of treatment, producing high volumes of high-quality sperm during several weeks. In addition, sperm cryopreservation protocols using different extenders, cryoprotectants and cooling and thawing times have been previously described for European eel. Here, we show that Tanaka¿s extender solution can be directly used for fertilization or for cryopreservation, making unnecessary the usage of different types of solutions and dilutions. Furthermore, the use of methanol as a cryoprotectant makes this protocol easy to use as methanol has low toxicity and does not activate the sperm. The sperm does not need to be cryopreserved immediately after the addition of the cryoprotectant, and it can be used long after being thawed. Moreover, sperm motility is still high after thawing although it is lower than that of fresh sperm. The aim of this work is to show the best available protocol for European eel handling, maturation, and sperm cryopreservation.
- Published
- 2018
24. Testing cryopreserved European eel sperm for hybridization (A. japonica × A. anguilla)
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministry of Education, Culture, Sports, Science and Technology, Japón, Tokyo University of Agriculture and Technology, Mohamed bin Zayed Species Conservation Fund, National Research, Development and Innovation Office, Hungría, European Commission, European Social Fund, Japan Society for the Promotion of Science, Müller, T., Matsubara, H., Kubara, Y., Horváth, Ákos, Kolics, B., Taller, J., Stéger, V., Kovács, B., Horváth, L., Asturiano Nemesio, Juan Francisco, Peñaranda, D.S., Urbanyi, B., Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministry of Education, Culture, Sports, Science and Technology, Japón, Tokyo University of Agriculture and Technology, Mohamed bin Zayed Species Conservation Fund, National Research, Development and Innovation Office, Hungría, European Commission, European Social Fund, Japan Society for the Promotion of Science, Müller, T., Matsubara, H., Kubara, Y., Horváth, Ákos, Kolics, B., Taller, J., Stéger, V., Kovács, B., Horváth, L., Asturiano Nemesio, Juan Francisco, Peñaranda, D.S., and Urbanyi, B.
- Abstract
[EN] The objective of this study was to assess impact of cryopreserved European eel sperm and Japanese eel native sperm on early fertilization, hatch, survival, and malformation rates of larvae, as well as develop molecular techniques to distinguish different eel species. Eggs from Japanese eel females (Anguilla japonica) were artificially fertilized with sperm of Japanese eel males and cryopreserved sperm from European eel (A. anguilla, extender was modified Tanaka solution and methanol as cryoprotectant). There were no statistical differences (p¿>¿0.05) among the measured parameters such as fertilization, hatch and survival after 10 days post-hatch rates due to large individual differences. The malformation rate of larvae compared to the hatching rate was higher in cryopreserved groups than in the control indicating that the methodology needs further refinement. Genetic analyses (PCR-RFLP, PCR-HRM) proved a clear result in the detection of paternal contribution in hybridization between the Japanese and the European eel and applied PCR-HRM method is a quick and cost effective tool to identify illegally imported A. anguilla at the glass eel stage, which can be transported from Europe to Asia.
- Published
- 2018
25. Long -term feeding with high plant protein based diets in gilthead seabream (Sparus aurata, L.) leads to changes in the inflammatory and immune related gene expression at intestinal level
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Educación, Universitat Politècnica de València, Estruch-Cucarella, Guillem, Collado, Mª Carmen, Monge-Ortiz, Raquel, Tomas-Vidal, A., Jover Cerdá, Miguel, Peñaranda, D.S., Perez Martinez, Gaspar, Martínez-Llorens, Silvia, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Educación, Universitat Politècnica de València, Estruch-Cucarella, Guillem, Collado, Mª Carmen, Monge-Ortiz, Raquel, Tomas-Vidal, A., Jover Cerdá, Miguel, Peñaranda, D.S., Perez Martinez, Gaspar, and Martínez-Llorens, Silvia
- Abstract
[EN] Background: In order to ensure sustainability of aquaculture production of carnivourous fish species such as the gilthead seabream (Sparus aurata, L.), the impact of the inclusion of alternative protein sources to fishmeal, including plants, has been assessed. With the aim of evaluating long-term effects of vegetable diets on growth and intestinal status of the on-growing gilthead seabream (initial weight = 129 g), three experimental diets were tested: a strict plant protein-based diet (VM), a fishmeal based diet (FM) and a plant protein-based diet with 15% of marine ingredients (squid and krill meal) alternative to fishmeal (VM+). Intestines were sampled after 154 days. Besides studying growth parameters and survival, the gene expression related to inflammatory response, immune system, epithelia integrity and digestive process was analysed in the foregut and hindgut sections, as well as different histological parameters in the foregut. Results: There were no differences in growth performance (p = 0.2703) and feed utilization (p = 0.1536), although a greater fish mortality was recorded in the VM group (p = 0.0141). In addition, this group reported a lower expression in genes related to pro-inflammatory response, as Interleukine-1 beta (il1 beta, p = 0.0415), Interleukine-6 (il6, p = 0.0347) and cyclooxigenase-2 (cox2, p = 0.0014), immune-related genes as immunoglobulin M (igm, p = 0.0002) or bacterial defence genes as alkaline phosphatase (alp, p = 0.0069). In contrast, the VM+ group yielded similar survival rate to FM (p = 0.0141) and the gene expression patterns indicated a greater induction of the inflammatory and immune markers (il1 beta, cox2 and igm). However, major histological changes in gut were not detected. Conclusions: Using plants as the unique source of protein on a long term basis, replacing fishmeal in aqua feeds for gilthead seabream, may have been the reason of a decrease in the level of different pro-inflammatory mediators (il1 beta, il6 and
- Published
- 2018
26. Using specific recombinant gonadotropins to induce spermatogenesis and spermiation in the European eel (Anguilla anguilla)
- Author
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Peñaranda, D.S., primary, Gallego, V., additional, Rozenfeld, C., additional, Herranz-Jusdado, J.G., additional, Pérez, L., additional, Gómez, A., additional, Giménez, I., additional, and Asturiano, J.F., additional
- Published
- 2018
- Full Text
- View/download PDF
27. Role of potassium and pH on the initiation of sperm motility in the European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, Ministerio de Economía, Industria y Competitividad, Generalitat Valenciana, Ministerio de Ciencia e Innovación, European Commission, Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Gallego Albiach, Victor, Asturiano Nemesio, Juan Francisco, Pérez Igualada, Luz María, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, Ministerio de Economía, Industria y Competitividad, Generalitat Valenciana, Ministerio de Ciencia e Innovación, European Commission, Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Gallego Albiach, Victor, Asturiano Nemesio, Juan Francisco, and Pérez Igualada, Luz María
- Abstract
[EN] The role of potassium from the seminal plasma and/or the activation media was examined by selectively removing from this media, and by testing the use of channel inhibitors and a K-ionophore. Sperm motility was measured using a CASA system, intracellular K+ and pH were measured by flow cytometry, and sperm head area was measured by ASMA: Automated Sperm Morphometry Analyses. Sperm motility was notably inhibited by the removal of K+ from the seminal plasma and by treatment with the K+ ionophore valinomycin. This therefore indicates that a reduction of K+ levels in the quiescent stage inhibits further motility. The normal decrease in sperm head area induced by seawater activation was altered by the removal of K+ from the seminal plasma, and an increase in the pH; in the quiescent stage was also induced. Intracellular pH (pH;) was quantitatively measured for the first time in European eel spermatozoa, being 7.2 in the quiescent stage and 7.1 post-activation. Intracellular and external pH levels influenced sperm motility both in the quiescent stage and at activation. The alkalinization of the pH; (by NH4Cl) inhibited sperm motility activation, while acidification (by Na-acetate) did not have any effect. Our results indicate that a pH gradient between the sperm cell and the seminal plasma is necessary for sperm motility activation. The presence of the ion K+ in the seminal plasma (or in the extender medium) is necessary in order to maintain sperm volume, intracellular pH and sperm motility.
- Published
- 2017
28. Feed restriction regime in a rabbit line selected for growth rate alters oocyte maturation manifested by alteration in msy2 gene expression
- Author
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Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Economía y Empresa, Ministerio de Economía, Industria y Competitividad, Naturil Alfonso, Carmen, Peñaranda, D.S., Vicente Antón, José Salvador, Marco-Jiménez, Francisco, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Generalitat Valenciana, Ministerio de Economía y Empresa, Ministerio de Economía, Industria y Competitividad, Naturil Alfonso, Carmen, Peñaranda, D.S., Vicente Antón, José Salvador, and Marco-Jiménez, Francisco
- Abstract
[EN] Young rabbit females selected for growth rate may have nutritional needs, which may not be met with the common practice of feed restriction during rearing in commercial rabbit production. The aim of this study was to analyse whether two different feeding programmes: ad libitum or restricted (130 g/day) feeding, applied in young rabbit females for 1 month at the end of rearing, could modulate the origin of ovulation process and the quality of the oocytes. At 16 weeks of age, 34 females were randomly assigned to restricted or ad libitum feeding, maintaining these conditions for a month. Then, in an initial experiment, transcriptional profiling of hypothalamus-hypophysis tissue was performed to assess failure to ovulate. In the second experiment, the gene expression analysis of some candidate genes related to oocytes quality was performed. Our results demonstrated that neither of the two feeding programmes modified the transcription of hypothalamus-hypophysis tissue, while the only differences in MSYR expression were found in in vivo mature oocytes ready for successful fertilization. Specifically, MSYR was over-expressed in oocytes from females fed ad libitum. MSYR is one of the most abundant proteins in the oocyte and has proven to be a key regulator of maternal RNA transcription and translation. This finding suggests that MSYR gene is a promising gene in our understanding of the relationship between high growth rate and reproductive performance decline.
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- 2017
29. The expression of nuclear and membrane estrogen receptors in the European eel throughout spermatogenesis
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Economía, Industria y Competitividad, Universitat Politècnica de València, Ministerio de Ciencia e Innovación, Generalitat Valenciana, MORINI, MARINA, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Tveiten, Helge, Lafont, Anne-Gaëlle, Dufour, Sylvie, Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Economía, Industria y Competitividad, Universitat Politècnica de València, Ministerio de Ciencia e Innovación, Generalitat Valenciana, MORINI, MARINA, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Tveiten, Helge, Lafont, Anne-Gaëlle, Dufour, Sylvie, Pérez Igualada, Luz María, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] Estradiol (E-2) can bind to nuclear estrogen receptors (ESR) or membrane estrogen receptors (GPER). While mammals possess two nuclear ESRs and one membrane GPER, the European eel, like most other teleosts, has three nuclear ESRs and two membrane GPERs, as the result of a teleost specific genome duplication. In the current study, the expression of the three nuclear ESRs (ESR1, ESR2a and ESR2b) and the two membrane GPERs (GPERa and GPERb) in the brain-pituitary-gonad (BPG) axis of the European eel was measured, throughout spermatogenesis. The eels were first transferred from freshwater (FW) to seawater (SW), inducing parallel increases in E2 plasma levels and the expression of ESRs. This indicates that salinity has a stimulatory effect on the E-2 signalling pathway along the BPG axis. Stimulation of sexual maturation by weekly injections of human chorionic gonadotropin (hCG) induced a progressive decrease in E-2 plasma levels, and different patterns of expression of ESRs and GPERs in the BPG axis. The expression of nuclear ESRs increased in some parts of the brain, suggesting a possible upregulation due to a local production of E-2. In the testis, the highest expression levels of the nuclear ESRs were observed at the beginning of spermatogenesis, possibly mediating the role of E2 as spermatogonia renewal factor, followed by a sharply decrease in the expression of ESRs. Conversely, there was a marked increase observed in the expression of both membrane GPERs throughout spermatogenesis, suggesting they play a major role in the final stages of spermatogenesis.
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- 2017
30. Nuclear and membrane progestin receptors in the European eel: characterization and expression in vivo through spermatogenesis
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, European Commission, Generalitat Valenciana, European Cooperation in Science and Technology, Ministerio de Economía, Industria y Competitividad, MORINI, MARINA, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Nourizadeh-Lillabadi, Rasoul, Lafont, Angael, Dufour, Sylvie, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, Pérez Igualada, Luz María, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, European Commission, Generalitat Valenciana, European Cooperation in Science and Technology, Ministerio de Economía, Industria y Competitividad, MORINI, MARINA, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Nourizadeh-Lillabadi, Rasoul, Lafont, Angael, Dufour, Sylvie, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, and Pérez Igualada, Luz María
- Abstract
[EN] Characterization of all the progestin receptor genes (PRs) found in the European eel has been performed. There were five membrane PRs (mPRs): mPR alpha (alpha), mPRAL1 (alpha-likel), mPRAL2 (alpha-like2), mPRy (gamma), mPR delta (delta) and two nuclear PRs (nPRs or PGRs): pgr1 and pgr2. In silico studies showed that the C and E(F) domains of Pgr are well conserved among vertebrates whereas the A/B domain is not. Phylogeny and synteny analyses suggest that eel duplicated pgr (pgr1 and pgr2) originated from the teleost-specific third whole genome duplication (3R). mPR phylogeny placed three eel mPRs together with the mPRce Glade, being termed mPRet, mPRAL1 and mPRAL2, while the other two eel mPRs clustered with mPRy and mPRS clades, respectively. The in vivo study showed differential expression patterns along the brain-pituitary-gonad axis. An increase in nPR transcripts was observed in brain (in pgrl) and pituitary (in pgrl and pgr2) through the spermatogenesis, from the spermatogonia B/spermatocyte stage to the spermiation stage. In the testis, mPRy, mPRS and pgr2 transcripts showed the highest levels in testis with A spermatogonia as dominant germ cell, while the highest mPRce, mPRAL1 and mPRAL2 transcripts were observed in testis from spermiating males, where the dominant germ cell were spermatozoa. Further studies should elucidate the role of both nuclear and membrane progestin receptors on eel spermatogenesis.
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- 2017
31. Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla)
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, National Research, Development and Innovation Office, Hungría, European Cooperation in Science and Technology, Ministry of Human Capacities, Hungría, Kása, E., Bernáth, G., Kollár, T., Zarski, D., Lujic, J., Marinovic, Z., Bokor, Z., Hegyi, Á., Urbányi, B., Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Horváth, Ákos, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, National Research, Development and Innovation Office, Hungría, European Cooperation in Science and Technology, Ministry of Human Capacities, Hungría, Kása, E., Bernáth, G., Kollár, T., Zarski, D., Lujic, J., Marinovic, Z., Bokor, Z., Hegyi, Á., Urbányi, B., Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, and Horváth, Ákos
- Abstract
[EN] Vitrification was successfully applied to the sperm of two fish species, the freshwater Eurasian perch (Perca fluviatilis) and marine European eel (Anguilla anguilla). Sperm was collected, diluted in species specific non-activating media and cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility of fresh and vitrified-thawed sperm was evaluated with computer-assisted sperm analysis (CASA). Additional sperm quality parameters such as sperm head morphometry parameters (in case of European eel) and fertilizing capacity (in case of Eurasian perch) were carried out to test the effectiveness of vitrification. The vitrification method for Eurasian perch sperm resulting the highest post-thaw motility (14 +/- 1.6%) was as follows: 1:5 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol + 15% propylene-glycol), cooling device: Cryotop, 2 mu l droplets, and for European eel sperm: dilution ratio 1:1, with 40% cryoprotectant (20% MeOH and 20% PG), and 10% FBS, cooling device: Cryotop, with 2 mu l of sperm suspension. Viable embryos were produced by fertilization with vitrified Eurasian perch sperm (neurulation: 2.54 +/- 1.67%). According to the ASMA analysis, no significant decrease in head area and perimeter of vitrified European eel spermatozoa were found when compared to fresh spermatozoa.
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- 2017
32. Transcript levels of the soluble sperm factor protein phospholipase C zeta 1 (PLCZ1) increase through induced spermatogenesis in European eel
- Author
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Morini, Marina Ange Marie, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Nourizadeh-Lillabadi, Rasoul, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, and Pérez Igualada, Luz María
- Subjects
BIOLOGIA ANIMAL ,Anguilla anguilla ,Teleost ,Reproduction ,Fertilization ,PRODUCCION ANIMAL ,Spermatozoa - Abstract
[EN] Activation at fertilization of the vertebrate egg is triggered by Ca2+ waves. Recent studies suggest the phospholipase C zeta (PLC zeta), a sperm-specific protein, triggers egg activation by an 1P3-mediated Ca2+ release and allow Ca2+ waves at fertilization. In the present study we cloned, characterized, and phylogenetically positioned the European eel PLC zeta (PLC zeta 1). It is 1521bp long, with 10 exons encoding an open reading frame of 506 amino acids. The amino acid sequence contains an EF-hand domain, X and Y catalytic domains, and a carboxy-terminal C2 domain, all typical of other PLC zeta orthologous. The tissue distribution was studied, and the gene expression was determined in testis during induced sexual maturation at three different thermal regimes. Also, brain and pituitary expression was studied through sex maturation at constant temperature. plc zeta was expressed in brain of male and female, in testis but not in ovaries. By first time in vertebrates, it is reported plc zeta 1 expression in the pituitary gland. Testis plc zeta 1 expression increased through spermatogenesis under all the thermal regimes, but being significantly elevated at lower temperatures. It was very low when testis contained only spermatogonia or spermatocytes, while maximum expression was found during spermiogenesis. These results support the hypothesis for an eel sperm-specific PLC zeta inducing egg activation, similarly to mammals and some teleosts, but different from some other teleost species, which express this protein in ovaries, but not in testes. (C) 2015 Elsevier Inc. All rights reserved., Funded by the SPERMOT project (Spanish Ministry of Science and Innovation, MICINN; AGL2010-16009). M.C. Vílchez has a predoctoral grant from UPV PAID Programme (2011-S2-02-6521), Marina Morini has a predoctoral grant from Generalitat Valenciana (Programa Grisolía, GRISOLIA/2012/006), Victor Gallego has a postdoctoral grant (UPV; PAID-10-14), and David S. Peñaranda was a contract cofinanced by MICINN and UPV (PTA2011-4948-I). Grants to attend meetings from COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE).
- Published
- 2015
33. The subpopulation pattern of eel sperm is affected by post-activation time, hormonal treatment and thermal regime
- Author
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Gallego Albiach, Victor, Vilchez Olivencia, Maria Carmen, Peñaranda, D.S., Pérez Igualada, Luz María, Herraez, María Paz, Asturiano Nemesio, Juan Francisco, and Martinez-Pastor, Felipe
- Subjects
BIOLOGIA ANIMAL ,European eel ,computer-aided sperm analysis ,PRODUCCION ANIMAL ,motility activation - Abstract
[EN] There has been a marked reduction in natural stocks of eels (genus Anguilla) over the past 60 years, and the culture of eels is still based on the capture of very large quantities of juveniles. It is necessary to close the life cycle in captivity in order to ease the pressure on wild populations. The aims of the present study were to evaluate sperm subpopulations (through cluster analysis of computer-aided sperm analysis data) in the European eel (Anguilla anguilla) and to assess the effects of motility acquisition time after activation (i.e. at 30, 60 and 90 s), the thermal regimen (i.e. 10 degrees C (T10) or 15 degrees C (T15) and up to 20 degrees C, or constant at 20 degrees C (T20)) and hormonal treatments (i.e. human chorionic gonadotropin (hCG), recombinant (r) hCG or pregnant mare serum gonadotropin (PMSG)) on these subpopulations. In all cases, we obtained three subpopulations of spermatozoa: low velocity and linear (S1); high velocity with low linearity (S2); and high velocity and linear (S3; considered high quality). Total motility and S1 were affected by acquisition time; thus, 30 s is recommended as the standard time for motility acquisition. When eels were kept at 20 degrees C (T20), motility data fitted quadratic models, with the highest motility and proportion of S3 between Weeks 8 and 12 after the first injection. Lower temperatures (T10, T15) delayed spermiation and the obtaining of high-quality spermatozoa (S3), but did not seem to alter the spermiation process (similar subpopulation pattern). Conversely, the hormonal treatments altered both the dynamics of the subpopulation pattern and the onset of spermiation (with PMSG delaying it). Total motility and the yield of S3 with the widely used hCG treatment varied throughout the spermiation period. However, using rhCG allowed us to obtain high-quality and constant motility for most of the study (Weeks 7-20), and the S3 yield was also higher overall (61.8 +/- 1.3%; mean +/- s.e.m.) and more stable over time than the other hormonal treatments (averaging 53.0 +/- 1.4%). Using T20 and rhCG would be more economical and practical, allowing us to obtain a higher number of S3 spermatozoa over an extended time., This study was funded by the European Community's 7th Framework Program under the Theme 2 'Food, Agriculture and Fisheries, and Bio-technology', grant agreement no. 245257 (PRO-EEL) and Generalitat Valenciana (ACOMP/2012/086). VG and MCV have predoctoral grants from the Spanish Ministry of Economy and Competitiveness (AGL2010-16009) and Universitat Politecnica de Valencia (UPV) PAID Program (2011-S2-02-6521), respectively. DSP was supported by a contract cofinanced by Ministry of Science and Innovation (MICINN) and UPV (PTA2011-4948-I). FM-P was supported by the Ramon y Cajal program (MICINN, RYC-2008-02560).
- Published
- 2015
34. Does vitrification alter the methylation pattern of OCT4 promoter in rabbit late blastocyst?
- Author
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Saenz-de-Juano, M.D., Peñaranda, D.S., Marco-Jiménez, F., and Vicente, J.S.
- Published
- 2014
- Full Text
- View/download PDF
35. Temperature modulates testis steroidogenesis in European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, European Commission, Ministerio de Economía y Competitividad, Generalitat Valenciana, Universitat Politècnica de València, Peñaranda, D.S., MORINI, MARINA, Tveiten, Helge, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Dirks, Ron, van den Thillart, Guido E.E.J.M., Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, European Commission, Ministerio de Economía y Competitividad, Generalitat Valenciana, Universitat Politècnica de València, Peñaranda, D.S., MORINI, MARINA, Tveiten, Helge, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Dirks, Ron, van den Thillart, Guido E.E.J.M., Pérez Igualada, Luz María, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] This study evaluates the effects of temperature on hCG-induced spermatogenesis in European eel (Anguilla anguilla), subjected to three thermal regimes: T10: 10 degrees C (first 4 weeks), 15 degrees C (next 3 weeks) and 20 degrees C (last 6 weeks); T15: 15 degrees C (first 4 weeks) and 20 degrees C (last 9 weeks); and T20: constant 20 degrees C for the duration of the experiment. At 10 degrees C, maturation stopped in the A spermatogonial stage (SPG1), and no further maturation was observed until the temperature was >= 15 degrees C. With the aim of explaining these results, the influence of temperature on steroidogenic enzyme gene expression and steroid synthesis was tested. The initial synthesis of androgens (T and 11-KT) increased at SPG1, and was not influenced by temperature. Likewise, the gene expression of the steroidogenic enzymes linked to androgen synthesis (aacyp11a1, aacyp17-I and aa11 beta HSD) also increased at SPG1. In contrast, no correlation was seen between the increase in E2 and the aacyp19a1 gene expression peak in the testes, with E2 increasing as a consequence of the seawater acclimation carried out before hormonal treatment, and peaking the aacyp19a1 gene expression at B spermatogonial stage (SPG2).Aacyp21 gene expression was also higher at SPG2, and this stage was only reached when the rearing temperature was >= 15 degrees C. In conclusion, androgen synthesis is not dependent on temperature, but further maturation requires higher temperatures in order to induce a change in the steroidogenic pathway towards estrogen and progestin synthesis. This study demonstrates that temperature plays a crucial role in European eel maturation, even perhaps controlling gonad development during the reproductive migration. (C) 2016 Elsevier Inc. All rights reserved.
- Published
- 2016
36. Sodium affects the sperm motility in the European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, Generalitat Valenciana, European Cooperation in Science and Technology, Universitat Politècnica de València, Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Gallego Albiach, Victor, Asturiano Nemesio, Juan Francisco, Pérez Igualada, Luz María, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, Generalitat Valenciana, European Cooperation in Science and Technology, Universitat Politècnica de València, Vilchez Olivencia, Maria Carmen, MORINI, MARINA, Peñaranda, D.S., Gallego Albiach, Victor, Asturiano Nemesio, Juan Francisco, and Pérez Igualada, Luz María
- Abstract
[EN] The role of seminal plasma sodium and activation media sodium on sperm motility was examined by selectively removing the element from these two media, in European eel sperm. Sperm size (sperm head area) was also measured using an ASMA (Automated Sperm Morphometry Analyses) system, in the different conditions. Intracellular sodium [Na+](i) was quantitatively analyzed by first time in the spermatozoa from a marine fish species. Measurement of [Na+](i) was done before and after motility activation, by Flow Cytometry, using CoroNa Green AM as a dye. Sperm motility activation induced an increase in [Na+](i) from 96.72 mM in quiescent stage to 152.21 mM post-activation in seawater. A significant decrease in sperm head area was observed post activation in seawater. There was a notable reduction in sperm motility when sodium was removed from the seminal plasma, but not when it was removed from the activation media. Sodium removal was also linked to a significant reduction in sperm head area in comparison to the controls. Our results indicate that the presence of the ion Na+ in the seminal plasma (or in the extender medium) is necessary for the preservation of sperm motility in European eel, probably because it plays a role in maintaining an appropriate sperm cell volume in the quiescent stage of the spermatozoa. (C) 2016 Elsevier Inc All rights reserved.
- Published
- 2016
37. Role of calcium on the initiation of sperm motility in the European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, Ministerio de Ciencia e Innovación, Pérez Igualada, Luz María, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, MORINI, MARINA, Peñaranda, D.S., Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, Ministerio de Ciencia e Innovación, Pérez Igualada, Luz María, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, MORINI, MARINA, Peñaranda, D.S., and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] Sperm from European eel males treated with hCG(rec), was washed in a calcium free extender, and sperm motility was activated both in the presence (seawater, SW) and in the absence of calcium (NaCI + EDTA), and treated with calcium inhibitors or modulators. The sperm motility parameters were evaluated by a computer-assisted sperm analysis (CASA) system, and changes in the [Ca2+](i) fluorescence (and in [Na+](i) in some cases) were evaluated by flow cytometry. After sperm motility was activated in a medium containing Ca2+ (seawater, SW) the intracellular fluorescence emitted by Ca2+ increased 4-6-fold compared to the levels in quiescent sperm. However, while sperm activation in a Ca-free media (NaCI + EDTA) resulted in a percentage of motility similar to seawater, the [Ca2+](i) levels did not increase at all. This result strongly suggests that increasing [Ca2+](i) is not a pre-requisite for the induction of sperm motility in European eel sperm. Several sperm velocities (VCL, VSL, VAP) decreased when sperm was activated in the Ca-free activator, thus supporting the theory that Ca2+ has a modulatory effect on sperm motility. The results indicate that a calcium/sodium exchanger (NCX) which is inhibited by bepridil and a calcium calmodulin kinase (inhibited by W-7), are involved in the sperm motility of the European eel. Our results indicate that the increase in [Ca2+](i) concentrations during sperm activation is due to an influx from the external medium, but, unlike in most other species, it does not appear to be necessary for the activation of motility in European eel sperm. (c) 2015 Elsevier Inc All rights reserved.
- Published
- 2016
38. Identification of the major proteins present in the seminal plasma of European eel, and how hormonal treatment affects their evolution. Correlation with sperm quality
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, European Cooperation in Science and Technology, Ministerio de Economía y Competitividad, Universitat Politècnica de València, Vilchez Olivencia, Maria Carmen, D. Pla, Gallego Albiach, Victor, Sanz, Libia, Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Calvete, Juan J., Peñaranda, D.S., Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural - Escola Tècnica Superior d'Enginyeria Agronòmica i del Medi Natural, European Cooperation in Science and Technology, Ministerio de Economía y Competitividad, Universitat Politècnica de València, Vilchez Olivencia, Maria Carmen, D. Pla, Gallego Albiach, Victor, Sanz, Libia, Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Calvete, Juan J., and Peñaranda, D.S.
- Abstract
[EN] By first time, 2DE protein profile of European eel seminal plasma has been determined. 14 different proteins corresponding to 9 major families were identified in seminal plasma, through hormonal treatment. Some of them play a part in sperm maturation, including carbonic anhydrase which is responsible for modulating the pH of seminal plasma, and warm temperature acclimation protein, which may play an important role in the final maturation of this species, due to the warm temperature of their spawning ground (in the Sargasso Sea). Sperm samples were classified into three motility categories depending on the percentage of motile cells, I: 0-25%, II: 25-50% and 111: >50%. Different protein profiles were observed depending on the sperm motility categories, specifically, with the apolipoproteins and complement C3. Higher numbers of proteins from the apolipoprotein family were registered at lower motilities; whereas the complement C3-like family was higher in the samples with the highest percentage of motile cells. These results suggest that the proteins linked to the transportation of lipids (apolipoprotein) and to the immune system (complement C3) may carry out their functions at different stages of spermatogenesis. Using SDS-PAGE analysis, 13 bands were identified, most of which migrated between 20 to 60 kDa. In the last weeks of treatment significant increases were observed in the percentage of motile spermatozoa, curvilinear velocity and beat cross frequency. This improvement in sperm quality coincided with a higher amount of proteins located at 19 KDa, therefore, this protein could be involved in sperm motility of the European eel. (C) 2016 Published by Elsevier Inc.
- Published
- 2016
39. Temperature modulates the vitellogenesis progression in European eel
- Author
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Mazzeo, Ilaria, Peñaranda, D.S., Gallego Albiach, Victor, Baloche, Sylvie, Nourizadeh-Lillabadi, Rasoul, Tveiten, H., Dufour, Sylvie, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, and Pérez Igualada, Luz María
- Subjects
endocrine system ,BIOLOGIA ANIMAL ,Anguilla anguilla ,Sex steroids ,Sex maturation ,PRODUCCION ANIMAL ,Thermal regime ,cyp19a1 - Abstract
[EN] Wild female European eels were matured with CPE (carp pituitary extract) under three thermal regimes, two of which were variable (T10–15 and T15–18, moving from 10 to 15 °C and from 15 to 18 °C, respectively) and one constant, at 18 °C (T18). Before and during hormonal treatment, the eels were sampled and biometric measurements were taken. Immunoassays of sex steroids and vitellogenin were performed, as well as qPCR analyses of gene expression (ovarian cyp19a1) and ovarian histology. Prior to the hormonal treatment, the silver eels which had been maintained at 18 °C showed lower 11-KT and E2 plasma levels compared to those maintained at 10 °C. In addition, in the early vitellogenic stage, the androgen and cyp19a1 levels were lower at 18 °C than at 10 °C. Both these results and the positive correlations found between GSI and 11-KT (at the PV stage) and between oocyte diameter and cyp19a1 levels (in the EV stage) suggest that early ovarian development is facilitated at low temperatures. Vitellogenesis was induced by CPE in all the thermal groups, but progression to the mid-vitellogenic stage was only observed after an accumulation of 900–1200 °D, at 15 or 18 °C, and progression to the late vitellogenic stage was only observed after an accumulation higher than 1300 °D, at 18 °C. Although temperature increased the rate of CPE-induced ovarian development, our results clearly indicate that this increase is not linear, but exponential, with acceleration in the increase of GSI at 18 °C from the mid-vitellogenic stage, or after an accumulation of 1300 °D. For the first time, a down-regulation of ovarian cyp19a1 caused by high temperatures in CPE-treated eels was observed. These results demonstrate that temperature can modulate eel ovarian development both before and after exogenous hormonal stimulations, and this knowledge could be used to manipulate the timing of vitellogenesis progression under laboratory conditions, This study was funded by the European Community's 7th Framework Programme under the Theme 2 "Food, Agriculture and Fisheries, and Biotechnology", grant agreement no.245257 (PRO-EEL). Ilaria Mazzeo and Victor Gallego had predoctoral grants from Generalitat Valenciana and Spanish Ministry of Science and Innovation (MICINN), respectively. David S. Penaranda has a postdoc grant from UPV (CEI-01-10), mobility grants from UPV (PAID-00-11) and the Research Council of Norway (EJ/hsm IS-SIP, 2009) and also has been supported by a contract co-financed by MICINN and UPV (PTA2011-4948-I). F.-A. Weltzien received funding from the Norwegian University of Life Sciences.
- Published
- 2014
40. Relationship between sperm quality parameters and fatty acid composition of the muscle, liver and testis of European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Ciencia e Innovación, Generalitat Valenciana, Universitat Politècnica de València, European Cooperation in Science and Technology, Baeza Ariño, Rosa, Mazzeo, Ilaria, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Peñaranda, D.S., Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Ciencia e Innovación, Generalitat Valenciana, Universitat Politècnica de València, European Cooperation in Science and Technology, Baeza Ariño, Rosa, Mazzeo, Ilaria, Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Peñaranda, D.S., Pérez Igualada, Luz María, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] This study looks at the correlations that fatty acids have with different tissues in the European eel (Anguilla anguilla L.) during hormonally-induced sexual maturation, with different sperm quality parameters. In order to evaluate the different dynamics of the use of fatty acids, a categorization of the results from each sperm quality parameter (volume, concentration, motility and velocity) was performed. Low and moderate correlations were observed between muscle tissue and some sperm quality parameters but no high correlations were found. Eicosapentaenoic add (20:5n3, EPA) in the liver seems to have a role in determining the volume of sperm produced. This can be explained by the fact that EPA is a major requirement in the early phases of sperm production (probably as a component of the spermatozoal membrane). In addition, the levels of alpha-linolenic acid (18:3-n3, ALA) and linoleic acid (18:2-n6, LA) in the liver decreased when sperm motility increased. In all the tissues, a negative correlation was observed between arachidonic acid (20:4n-6, ARA) and the different sperm velocity parameters. The fact that an increase in the consumption of ARA coincides with an increase in the speed of spermatozoa, highlights the important role that this fatty acid plays not only in sperm production, but also in sperm velocity. All this information could prove useful in the development of suitable broodstock diets to improve sperm quality and subsequently, the larval development of this species. (C) 2014 Elsevier Inc All rights reserved.
- Published
- 2015
41. The subpopulation pattern of eel sperm is affected by post-activation time, hormonal treatment and thermal regime
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Generalitat Valenciana, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Gallego Albiach, Victor, Vilchez Olivencia, Maria Carmen, Peñaranda, D.S., Pérez Igualada, Luz María, Herraez, María Paz, Asturiano Nemesio, Juan Francisco, Martinez-Pastor, Felipe, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Generalitat Valenciana, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Gallego Albiach, Victor, Vilchez Olivencia, Maria Carmen, Peñaranda, D.S., Pérez Igualada, Luz María, Herraez, María Paz, Asturiano Nemesio, Juan Francisco, and Martinez-Pastor, Felipe
- Abstract
[EN] There has been a marked reduction in natural stocks of eels (genus Anguilla) over the past 60 years, and the culture of eels is still based on the capture of very large quantities of juveniles. It is necessary to close the life cycle in captivity in order to ease the pressure on wild populations. The aims of the present study were to evaluate sperm subpopulations (through cluster analysis of computer-aided sperm analysis data) in the European eel (Anguilla anguilla) and to assess the effects of motility acquisition time after activation (i.e. at 30, 60 and 90 s), the thermal regimen (i.e. 10 degrees C (T10) or 15 degrees C (T15) and up to 20 degrees C, or constant at 20 degrees C (T20)) and hormonal treatments (i.e. human chorionic gonadotropin (hCG), recombinant (r) hCG or pregnant mare serum gonadotropin (PMSG)) on these subpopulations. In all cases, we obtained three subpopulations of spermatozoa: low velocity and linear (S1); high velocity with low linearity (S2); and high velocity and linear (S3; considered high quality). Total motility and S1 were affected by acquisition time; thus, 30 s is recommended as the standard time for motility acquisition. When eels were kept at 20 degrees C (T20), motility data fitted quadratic models, with the highest motility and proportion of S3 between Weeks 8 and 12 after the first injection. Lower temperatures (T10, T15) delayed spermiation and the obtaining of high-quality spermatozoa (S3), but did not seem to alter the spermiation process (similar subpopulation pattern). Conversely, the hormonal treatments altered both the dynamics of the subpopulation pattern and the onset of spermiation (with PMSG delaying it). Total motility and the yield of S3 with the widely used hCG treatment varied throughout the spermiation period. However, using rhCG allowed us to obtain high-quality and constant motility for most of the study (Weeks 7-20), and the S3 yield was also higher overall (61.8 +/- 1.3%; mean +/- s.e.m.) and more stable
- Published
- 2015
42. Exploring correlations between sex steroids and fatty acids and their potential roles in the induced maturation of the male European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Baeza Ariño, Rosa, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Tveiten, Helge, Pérez Igualada, Luz María, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Baeza Ariño, Rosa, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Tveiten, Helge, Pérez Igualada, Luz María, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] The present study was undertaken to evaluate the correlations between the fatty acids in the liver and testis and the plasma levels of the hormonal steroids used during eel spermatogenesis, in order to clarify the physiological roles fatty acids play in the spermatogenetic process. The stages of testis development (S1-S5) were assessed by histological observations in order to classify the different phases of hormonally-induced spermatogenesis and evaluate the possible relationships between the hormones and fatty acids in each stage. The highest plasma levels of 17 beta-estradiol (E2), testosterone (T) and 11-ketotestosterone (11KT) were found in S1, when spermatogonial proliferation occurs. A correlation was found between 17 alpha-20 beta-dihydroxy-4-pregnen-3- one (DHP) levels and some fatty acids during the proliferation and growing phases (S1-2), suggesting that DHP might modulate lipid metabolism in the liver during early spermatogenesis. The DHP levels increased significantly during the growing phase (S2) and remained at high levels throughout the subsequent development stages (S3-S5). Similar to results found in mammals, our results show that in the eel there are regulatory mechanisms, including eicosapentaenoic acid (20:5-n3, EPA) and docosahexaenoic acid (22:6-n3, DHA), which act as modulators in the synthesis of androgens, particularly during the final phase of sperm maturation. Our results suggest that the fact that EPA, ARA and DHA concentrations in the eel testis remain constant/stable during spermiation could be related to the subsequent union of the spermatozoa and the egg. The findings from this research provide new insights for further studies about the possible effect of steroids on desaturase activity and highlight the importance of the effect of lipid metabolism during male eel spermatogenesis. (C) 2014 Elsevier B.V. All rights reserved.
- Published
- 2015
43. Effect of the probiotic Lactobacillus rhamnosus on the expression of genes involved in European eel spermatogenesis
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Universitat Politècnica de València, European Cooperation in Science and Technology, Ministerio de Ciencia e Innovación, Vilchez Olivencia, Maria Carmen, Santangeli, Stefania, Maradonna, Francesca, Gioacchini, Giorgia, Verdenelli, Cristina, Gallego Albiach, Victor, Peñaranda, D.S., Tveiten, Helge, Pérez Igualada, Luz María, Carnevali, Oliana, Asturiano Nemesio, Juan Francisco, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, European Commission, Universitat Politècnica de València, European Cooperation in Science and Technology, Ministerio de Ciencia e Innovación, Vilchez Olivencia, Maria Carmen, Santangeli, Stefania, Maradonna, Francesca, Gioacchini, Giorgia, Verdenelli, Cristina, Gallego Albiach, Victor, Peñaranda, D.S., Tveiten, Helge, Pérez Igualada, Luz María, Carnevali, Oliana, and Asturiano Nemesio, Juan Francisco
- Abstract
[EN] Positive effects of probiotics on fish reproduction have been reported in several species. In the present study, 40 male European eels were weekly treated with recombinant hCG for 9 weeks and with three different concentrations (10(3), 10(5), and 10(6) CFU/mL) of probiotic Lactobacillus rhamnosus IMC 501 (Sinbyotec, Italy). The probiotics were daily added to the water from the sixth week of the hCG treatment. Males from the treated and control groups were sacrificed after 1, 2, and 3 weeks of probiotic treatment (seventh ninth weeks of hCG treatment); at this point, sperm and testis samples were also collected. Sperm volume was estimated, and motility was analyzed by computer-assisted sperm analysis software. Alternations in transcription of specific genes involved in reproductive process such as activin, androgen receptors alpha and beta (ar alpha and ar beta), progesterone receptor 1 (pr1), bone morphogenetic protein 15 (bmp15), and FSH receptor (fshr) were analyzed in the testis. After 2 weeks of probiotic treatment, sperm production and sperm motility parameters (percentage of motile cells and percentage of straight-swimming spermatozoa) were increased in the European eel treated with 105 CFU/mL compared to controls or to the other probiotic doses. These changes were associated with increases in messenger RNA expression of activin, ar alpha, ar beta, pr1, and fshr. Conversely, after 3 weeks, activin and pr1 expression decreased. No significant changes were observed on bmp15 expression throughout the duration of the treatment with 10(5) CFU/mL dose. The lowest and highest probiotic dose (10(3) and 10(6) CFU/mL, respectively) inhibited the transcription of all genes along all the experiment, except for ar alpha and ar beta after 1 week of probiotic treatment when compared to controls. The changes observed by transcriptomic analysis and the sperm parameters suggest that a treatment with L rhamnosus at 10(5) CFU/mL for 2 weeks could improve spermatogenesis proc
- Published
- 2015
44. Transcript levels of the soluble sperm factor protein phospholipase C zeta 1 (PLCZ1) increase through induced spermatogenesis in European eel
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Generalitat Valenciana, European Commission, European Cooperation in Science and Technology, Morini, Marina Ange Marie, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Nourizadeh-Lillabadi, Rasoul, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, Pérez Igualada, Luz María, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Ministerio de Ciencia e Innovación, Universitat Politècnica de València, Generalitat Valenciana, European Commission, European Cooperation in Science and Technology, Morini, Marina Ange Marie, Peñaranda, D.S., Vilchez Olivencia, Maria Carmen, Gallego Albiach, Victor, Nourizadeh-Lillabadi, Rasoul, Asturiano Nemesio, Juan Francisco, Weltzien, Finn-Arne, and Pérez Igualada, Luz María
- Abstract
[EN] Activation at fertilization of the vertebrate egg is triggered by Ca2+ waves. Recent studies suggest the phospholipase C zeta (PLC zeta), a sperm-specific protein, triggers egg activation by an 1P3-mediated Ca2+ release and allow Ca2+ waves at fertilization. In the present study we cloned, characterized, and phylogenetically positioned the European eel PLC zeta (PLC zeta 1). It is 1521bp long, with 10 exons encoding an open reading frame of 506 amino acids. The amino acid sequence contains an EF-hand domain, X and Y catalytic domains, and a carboxy-terminal C2 domain, all typical of other PLC zeta orthologous. The tissue distribution was studied, and the gene expression was determined in testis during induced sexual maturation at three different thermal regimes. Also, brain and pituitary expression was studied through sex maturation at constant temperature. plc zeta was expressed in brain of male and female, in testis but not in ovaries. By first time in vertebrates, it is reported plc zeta 1 expression in the pituitary gland. Testis plc zeta 1 expression increased through spermatogenesis under all the thermal regimes, but being significantly elevated at lower temperatures. It was very low when testis contained only spermatogonia or spermatocytes, while maximum expression was found during spermiogenesis. These results support the hypothesis for an eel sperm-specific PLC zeta inducing egg activation, similarly to mammals and some teleosts, but different from some other teleost species, which express this protein in ovaries, but not in testes. (C) 2015 Elsevier Inc. All rights reserved.
- Published
- 2015
45. Impact of Fishmeal Replacement in Diets for Gilthead Sea Bream (Sparus aurata) on the Gastrointestinal Microbiota Determined by Pyrosequencing the 16S rRNA Gene
- Author
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Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, Estruch, Guillem, Collado Amores, María Carmen, Peñaranda, D.S., Tomas-Vidal, A., Jover Cerdá, Miguel, Pérez-Martínez, G., Martínez-Llorens, Silvia, Universitat Politècnica de València. Instituto de Ciencia y Tecnología Animal - Institut de Ciència i Tecnologia Animal, Universitat Politècnica de València. Departamento de Ciencia Animal - Departament de Ciència Animal, Universitat Politècnica de València, Estruch, Guillem, Collado Amores, María Carmen, Peñaranda, D.S., Tomas-Vidal, A., Jover Cerdá, Miguel, Pérez-Martínez, G., and Martínez-Llorens, Silvia
- Abstract
[EN] Recent studies have demonstrated the impact of diet on microbiota composition, but the essential need for the optimization of production rates and costs forces farms and aquaculture production to carry out continuous dietary tests. In order to understand the effect of total fishmeal replacement by vegetable-based feed in the sea bream (Sparus aurata), the microbial composition of the stomach, foregut, midgut and hindgut was analysed using high-throughput 16S rDNA sequencing, also considering parameters of growth, survival and nutrient utilisation indices.A total of 91,539 16S rRNA filtered-sequences were analysed, with an average number of 3661.56 taxonomically assigned, high-quality sequences per sample. The dominant phyla throughout the whole gastrointestinal tract were Actinobacteria, Protebacteria and Firmicutes. A lower diversity in the stomach in comparison to the other intestinal sections was observed. The microbial composition of the Recirculating Aquaculture System was totally different to that of the sea bream gastrointestinal tract. Total fishmeal replacement had an important impact on microbial profiles but not on diversity. Streptococcus (p-value: 0.043) and Photobacterium (p-value: 0.025) were highly represented in fish fed with fishmeal and vegetable-meal diets, respectively. In the stomach samples with the vegetable diet, reads of chloroplasts and mitochondria from vegetable dietary ingredients were rather abundant. Principal Coordinate Analysis showed a clear differentiation between diets in the microbiota present in the gut, supporting the presence of specific bacterial consortia associated with the diet.Although differences in growth and nutritive parameters were not observed, a negative effect of the vegetable diet on the survival rate was determined. Further studies are required to shed more light on the relationship between the immune system and sea bream gastrointestinal tract microbiota and should consider the modulation of the microbiot
- Published
- 2015
46. Up-regulation of insulin-like growth factor I and uteroglobin in in vivo-developed parthenogenetic embryos
- Author
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Naturil Alfonso, Carmen, Vicente Antón, José Salvador, Peñaranda, D.S., and Marco Jiménez, Francisco
- Subjects
animal structures ,Mouse ,Parthenogenesis ,Activation ,Gene Expression Regulation, Developmental ,PRODUCCION ANIMAL ,Vitro ,Developmental competence ,Rabbit oocytes ,Induction ,Up-Regulation ,BIOLOGIA ANIMAL ,Pluripotent stem cells ,Differentiation ,embryonic structures ,Bovine blastocysts ,Oocytes ,Animals ,Uteroglobin ,Female ,Gene-expression ,RNA, Messenger ,Rabbits ,Insulin-Like Growth Factor I - Abstract
[EN] Contents Parthenote embryos are being considered as an alternative source of embryonic stem cells. However, as there is still a dearth of knowledge of this kind of embryos, a better understanding of their biology is needed for their application. In this work, we studied the differences and similarities between parthenotes and normal embryos at the blastocyst stage in vivo developed. We analysed the expression of factor OCT-4, vascular endothelial growth factor (VEGF), insulin-like growth factor I (IGF-I) and uteroglobin (UG) by real-time PCR. To do so, oocytes were recovered and after activation procedure were transferred by ventral middle laparoscopy to receptive does to undergo completely in vivo development. Does were slaughtered 6 days post-ovulation induction, and parthenote and normal embryos were recovered for mRNA expression analysis. Our results reported that parthenotes and normal embryos showed similar mRNA expression for OCT-4 and VEGF. However, IGF-I and UG showed to be over-expressed in parthenote embryos. Thus, our study highlights that despite the in vivo development of parthenotes, they still seem to have an altered expression and, therefore, to be different to normal embryos. The altered expression pattern of parthenote embryos suggests that these embryos should be studied carefully before future application., This work was supported by Generalitat Valenciana research programme (Prometeo 2009/125). The authors thank Neil Macowan Language Services for revising the English version of the manuscript.
- Published
- 2012
47. mRNA Expression in Rabbit Blastocyst and Endometrial Tissue of Candidate Gene Involved in Gestational Losses
- Author
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Llobat, L., Marco Jiménez, Francisco, Peñaranda, D.S., Thieme, R., Navarrete, A., and Vicente Antón, José Salvador
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Abortion, Veterinary ,PRODUCCION ANIMAL ,Weight Gain ,Receptor, IGF Type 2 ,Endometrium ,Blastocyst ,BIOLOGIA ANIMAL ,Gene Expression Regulation ,Pregnancy ,Animals ,Female ,Genetic Predisposition to Disease ,RNA, Messenger ,Rabbits ,Gestation ,Biomarkers ,Signal Transduction - Abstract
[EN] Gestation is a complex process that involves different growth factors, cytokines and adhesion proteins related with embryo development, cellular differentiation and proliferation, embryo-endometrium interaction, angiogenesis, maternal-embryonic recognition and growth development of placenta and embryos. In this study, we examine pre-implantational (at 6days of gestation) and gestational (at 12days and total from ovulation to birth) losses in two rabbit lines selected by different criteria (post-weaning daily gain and litter size) and the pattern of a set of candidate transcripts, at 6days of gestation, related with embryo development and implantation process, such as Oct-4, epidermal growth factor receptor 3 (erbB3), Transforming Growth Factor ß2, Vascular Endothelial Growth Factor and Interferon ¿ and related with insulin-like growth factors signalling as insulin growth factors I and II and their receptors in rabbit blastocysts and endometrial tissue. Similar pre-implantational losses were obtained in both lines. However, the gestational losses of the line selected by post-weaning daily gain clearly mirrored an increase in losses by 50% at 12days and at birth (22.4 vs 9.5 and 50.2 vs 25.4, respectively, between line selected by post-weaning daily gain and line selected by litter size). In blastocysts and endometrial tissue at 6days of gestation qRT-PCR assays indicated that the mean insulin-like growth factor (IGF)-IIR mRNA expression was down-regulated in line selected by post-weaning daily gain. Dysregulation of the IGF-IIR could be potential reasons for induced gestational losses. We conclude that IGF-IIR gene expression in blastocyst and endometrial tissue at 6th day of gestation tends to decline in line selected by post-weaning daily gain. The functional significance related with gestational losses is uncertain. © 2011 Blackwell Verlag GmbH., We are grateful to the Researcher Personal Training Programme from Valencia Polytechnic University. This work was supported by the Spanish Research Project (CICYT AGL2008-03274). The authors thank Neil Macowan Language Services for revising the English version of the manuscript.
- Published
- 2012
48. effect of embryonic genotype on reference gene selection for RT-qPCR normalization
- Author
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Llobat, L., Marco Jiménez, Francisco, Peñaranda, D.S., Saenz de Juano Ribes, María de los Desamparados, and Vicente Antón, José Salvador
- Subjects
Vascular Endothelial Growth Factor A ,Genotype ,Receptor, ErbB-3 ,Gene Expression ,Expression ,Relative quantification ,PRODUCCION ANIMAL ,Real-Time Polymerase Chain Reaction ,Histones ,Interferon-gamma ,Transforming Growth Factor beta2 ,In vitro ,Validation ,Animals ,RNA, Messenger ,Time quantitative PCR ,Housekeeping genes ,Gene Expression Profiling ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Embryo, Mammalian ,Bovine preimplantation embryos ,Transcription PCR data ,Real time ,BIOLOGIA ANIMAL ,Blastocyst ,Oocytes ,Rabbits ,Octamer Transcription Factor-3 - Abstract
[EN] Contents To obtain more reliable results with biological significance, it requires data normalization using an appropriate internal control gene. In rabbits, there are classic stable reference genes that have been identified for normalization in oocytes and pre-implantation stage embryos. However, effects of embryonic genotype on reference gene selection have not been elucidated. The aim of this study was to test (i) the stability of mRNA transcription level for histone (H2afz) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) genes in rabbit blastocysts from two lines selected by different criteria (litter size and post-weaning daily weight gain) and (ii) its influence on biological significance examined by means of a set of embryonic transcripts, such as POU5F1 (Oct-4), epidermal growth factor receptor (erbB3), transforming growth factor-beta2, vascular endothelial growth factor and gamma interferon (Ifn-gamma). The geNorm, NormFinder and BestKeeper programs showed similar results, pointing out that H2afz and GAPDH were the most stable reference genes in rabbits selected on litter size at weaning. Moreover, our study revealed that embryonic genotype affected target gene expression when a single reference gene was used to analyse mRNA expression in blastocysts. Results showed that GAPDH gene is better than H2afz for gene expression studies of both embryo genotypes. A normalization factor derived from H2afz and GAPDH is likely to be appropriate when RT-qPCR was performed in rabbit embryos with different genotypes., We are grateful to the Researcher Personal Training Programme from Valencia Polytechnic University. This work was supported by the Spanish Research Project (CICYT AGL2008-03274) and Generalitat Valenciana Research Programme (Prometeo 2009/125). The authors thank Neil Macowan Language Services for revising the English version of the manuscript.
- Published
- 2011
49. Relationship between sperm quality parameters and the fatty acid composition of the muscle, liver and testis of European eel
- Author
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Baeza, R., primary, Mazzeo, I., additional, Vílchez, M.C., additional, Gallego, V., additional, Peñaranda, D.S., additional, Pérez, L., additional, and Asturiano, J.F., additional
- Published
- 2015
- Full Text
- View/download PDF
50. Exploring correlations between sex steroids and fatty acids and their potential roles in the induced maturation of the male European eel
- Author
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Baeza, R., primary, Peñaranda, D.S., additional, Vílchez, M.C., additional, Tveiten, H., additional, Pérez, L., additional, and Asturiano, J.F., additional
- Published
- 2015
- Full Text
- View/download PDF
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