Browsing by Subject "Reproductive fluids"
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- PublicationOpen AccessReproductive fluids reduce the rate of telomer shortening during the first year of life in in vitro derived- pig and cattle(Springer Nature, 2022) Garcia-Cobarro, Claudia; Romar, Raquel; Ramis Vidal, Guillermo; Canovas, Sebastian; Gutiérrez-Adán, Alfonso; Coy, Pilar; FisiologíaThe shortening rate of telomere length in blood cells is a potential predictive tool for health and lifespan. It is unknown whether the differences in telomere shortening observed between individuals are genetically determined or whether early environmental factors produce such differences. In this study, we analyzed the relative length of leukocyte telomeres throughout the first year of life in two animal colonies of the bovine and porcine species. The animals were generated by artificial insemination (AI) (control group) or by in vitro production (IVP) and transfer of embryos, either using conventional culture media (C-IVP, with bovine serum albumin as the only protein source) or enriched culture media (ET-IVP, with reproductive fluids as an extra source of proteins). Blood samples obtained 3 days, one month (day 28-30), 4 months (day 120), and one year (day 365) after birth were used. Cellular DNA was extracted, purified, and quantified following standardized protocols. Subsequently, the telomere length was determined by real-time quantitative PCR, normalizing the telomere amplification values with respect to the amount of the constitutive gene Rn18S (relative quantification). Statistical analysis (one way ANOVA and Bonferroni in cattle, two-sample t-test for inhomogeneous variances in pig, Pearson correlation and regression analysis) of the relative telomere size with respect to age and animal group allowed us to observe the dynamics of shortening that occurs in both species. The results showed a progressive shortening in telomere length with age, with the greatest shortening occurring in cattle at 30 days and in pigs at 120 days. As for the speed of shortening, in cattle, it was observed that the C-IVP group suffered a greater shortening between days 30 and 120 compared to the AI and ET-IVP groups. In pigs, a greater reduction was also observed in the C-IVP group than in the ET-IVP group. The results in both species suggest that supplementation with reproductive fluids during IVP reduces the rate of shortening of telomere length during the early development of individuals.
- PublicationOpen AccessReproductive fluids, added to the culture media, contribute to minimizing phenotypical differences between in vitro-derived and artificial insemination-derived piglets(Cambridge University Press, 2022) París-Oller, Evelyne; Soriano-Úbeda, Cristina; Belda-Pérez, Ramsés; Sarriás-Gil, Lucía; Canha-Gouveia, Analuce; Gadea, Joaquín; Vieira, Luis Alberto; García-Vázquez, Francisco Alberto; Romar, Raquel; Cánivas, Sebastián; Coy, Pilar; FisiologíaThe addition of reproductive fluids (RF) to the culture media has shown benefits in different embryonic traits but its long-term effects on the offspring phenotype are still unknown. We aimed to describe such effects in pigs. Blood samples and growth parameters were collected from piglets derived from in vitro-produced embryos (IVP) with or without RF added in the culture media versus those artificially inseminated (AI), from day 0 to month 6 of life. An oral glucose tolerance test was performed on day 45 of life. We show here the first comparative data of the growth of animals produced through different assisted reproductive techniques, demonstrating differences between groups. Overall, there was a tendency to have a larger size at birth and faster growth in animals derived from in vitro fertilization and embryo culture versus AI, although this trend was diminished by the addition of RFs to the culture media. Similarly, small differences in hematological indices and glucose tolerance between animals derived from AI and those derived from IVP, with a sex-dependent effect, tended to fade in the presence of RF. The addition of RF to the culture media could contribute to minimizing the phenotypical differences between the in vitro-derived and AI offspring, particularly in males.
- PublicationOpen AccessReproductive fluids, used for the in vitro production of pig embryos, result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM(BMC, 2021) París-Oller, E.; Navarro-Serna, S.; Soriano-Úbeda, C.; Lopes, J.S.; Matás, C.; Ruiz, S.; Latorre, L.; Romar, R.; Cánovas, S.; Coy, P.; López Albors, Octavio Miguel; FisiologíaBackground: In vitro embryo production (IVP) and embryo transfer (ET) are two very common assisted reproductive technologies (ART) in human and cattle. However, in pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow. In addition, the potential impact of these two ART on the health of the offspring is unknown. We investigated here if the use of a modified IVP system, with natural reproductive fluids (RF) as supplements to the culture media, combined with a minimally invasive surgery to perform ET, affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits. Results: The blastocyst rates obtained by both in vitro systems, conventional (C-IVP) and modified (RF-IVP), were similar. Pregnancy and farrowing rates were also similar. However, when compared to in vivo control (artificial insemination, AI), litter sizes of both IVP groups were lower, while placental efficiency was higher in AI than in RF-IVP. Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI, but not for RF-IVP group. Conclusions: The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring.
- PublicationOpen AccessReproductive fluids, used for the in vitro production of pig embryos, result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM.(BMC, 2021-02-15) París-Oller, E.; Navarro-Serna, S.; Soriano-Ubeda, C.; Lopes, J.S.; Matás, C.; Ruiz, S.; Cánovas, S.; Coy, P.; Romar, R.; Latorre Reviriego, Rafael Manuel; López Albors, Octavio Miguel; Fisiología; Anatomía y Anatomía Patológica ComparadasBackground: In vitro embryo production (IVP) and embryo transfer (ET) are two very common assisted reproductive technologies (ART) in human and cattle. However, in pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow. In addition, the potential impact of these two ART on the health of the offspring is unknown. We investigated here if the use of a modified IVP system, with natural reproductive fluids (RF) as supplements to the culture media, combined with a minimally invasive surgery to perform ET, affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits. Results: The blastocyst rates obtained by both in vitro systems, conventional (C-IVP) and modified (RF-IVP), were similar. Pregnancy and farrowing rates were also similar. However, when compared to in vivo control (artificial insemination, AI), litter sizes of both IVP groups were lower, while placental efficiency was higher in AI than in RF-IVP. Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI, but not for RF-IVP group. Conclusions: The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring. Keywords: Assisted reproductive technologies, Embryo transfer, In vitro embryo production, Placenta, Reproductive fluids
- PublicationOpen AccessSpontaneous and ART-induced large offspring syndrome: similarities and differences in DNA methylome(Taylor and Francis Group, 2022-05-03) Yahan Li; Sena Lopes, Jordana; Coy-Fuster, Pilar; Melissa Rivera, Rocío; Didáctica y Organización EscolarLarge/abnormal offspring syndrome (LOS/AOS) is a congenital overgrowth syndrome reported in ruminants produced by assisted reproduction (ART-LOS) which exhibit global disruption of the epigenome and transcriptome. LOS/AOS shares phenotypes and epigenotypes with the human congenital overgrowth condition Beckwith-Wiedemann syndrome. We have reported that LOS occurs spontaneously (SLOS); however, to date, no study has been conducted to determine if SLOS has the same methylome epimutations as ART-LOS. In this study, we performed whole- genome bisulphite sequencing to examine global DNA methylation in bovine SLOS and ART-LOS tissues. We observed unique patterns of global distribution of differentially methylated regions (DMRs) over different genomic contexts, such as promoters, CpG Islands, shores and shelves, as well as at repetitive sequences. In addition, we included data from two previous LOS studies to identify shared vulnerable genomic loci in LOS. Overall, we identified 320 genomic loci in LOS that have alterations in DNA methylation when compared to controls. Specifically, there are 25 highly vulnerable loci that could potentially serve as molecular markers for the diagnosis of LOS, including at the promoters of DMRT2 and TBX18, at the imprinted gene bodies of IGF2R, PRDM8, and BLCAP/NNAT, and at multiple CpG Islands. We also observed tissue-specific DNA methylation patterns between muscle and blood, and conservation of ART-induced DNA methyla-tion changes between muscle and blood. We conclude that as ART-LOS, SLOS is an epigenetic condition. In addition, SLOS and ART-LOS share similarities in methylome epimutations.
- PublicationOpen AccessThe impact of in vitro embryo production on placental and umbilical cord vascularization is minimized by the addition of reproductive fluids(Elsevier, 2023-09-15) Álvarez Martín, Úrsula; Coy, Pilar; Romar, Raquel; Párraga Ros, Ester; Seva Alcaraz, Juan; Anatomía y Anatomía Patológica ComparadasAnimals born from in-vitro-produced (IVP) embryos show changes in the placenta and umbilical cord vascularization. This study compares the placental and umbilical vascular morphometry in pigs (n = 19) born through artificial insemination (AI group) or after transfer of IVP embryos cultured with (RF-IVP group) or without (C-IVP group) reproductive fluids. The relationship between vascular parameters and animal growth during the first year of life was also analyzed. Samples were collected at birth, fixed, paraffin-embedded, cut in sections, stained, and photographed for vascular and morphometric analysis with ImageJ® and Slide Viewer®. The average daily weight gain was individually scored from birth to the first year of life. No differences were found in placental vascular morphometry among groups, except for the vascular area of small vessels (arterioles, venules, and small vessels) that was higher in the C-IVP group. Regarding the umbilical cord, the values for perimeter (AI: 26.40 ± 3.93 mm; IVP: 30.51 ± 4.74 mm), diameter (AI: 8.35 ± 1.01 mm; IVP: 10.26 ± 1.85 mm), area (AI: 43.18 ± 12.87; IVP: 56.61 ± 14.89 mm2), and Wharton's jelly area (AI: 36.86 ± 12.04 mm2; IVP 48.88 ± 12.80 mm2) were higher in IVP-derived than AI-derived animals, whereas arterial and venous morphometric data were similar between groups. A correlation study showed that placental and umbilical cord vascular phenotypes affect the further growth of pigs. In conclusion, assisted reproductive technologies impact small caliber vessels in the placenta and morphometric parameters in the umbilical cord. The addition of reproductive fluids in IVP-embryo contributes to reduce the differences with in vivo-derived animals.
- PublicationOpen AccessYear-Long Phenotypical Study of Calves Derived From Different Assisted-Reproduction Technologies(Frontiers Media, 2022-01-10) Lopes, Jordana S.; Soriano-Úbeda, Cristina; Navarro-Serna, Sergio; París-Oller, Evelyne; Canha-Gouveia, Analuce; Sarrias-Gil, Lucía; Cerón, José Joaquin; CoY, Pilar; Didáctica y Organización EscolarAssisted reproductive technologies play a major role in thecattle industry. An increasein the use ofin vitro-derived embryos is currently being seen around the globe. But theefficiency and quality of thein vitro-derived embryos are substandard when comparedto thein vivoproduction. Different protocols have been designed to overcome this issue,one of those being the use of reproductive fluids as supplementation to embryo culturemedia. In this study,in vitro-derived calves produced with reproductive fluids added totheir embryo production protocol were followed for the firstyear of life pairwise withtheirin vivocontrol, produced by artificial insemination (AI), and theirin vitrocontrol,produced with standard supplementation in embryo production. The objective was toassess if any differences could be found in terms of growth and development as well ashematological and biochemical analytes between the different systems. All the analysedvariables (physical, hematological, and biochemical) were within physiological range andvery similar between calves throughout the entire experiment. However, differences weremore evident between calves derived from standardin vitroproduction and AI. Weconcluded that the use of reproductive fluids as a supplementation to the embryo culturemedia results in calves with closer growth and development patterns to those born byAI than the use of bovine serum albumin as supplementation.