Vitrification Effects on the Transcriptome of in vivo-Derived Porcine Morulae

Cuello, C.; Martínez, Cristina A.; Cambra, Josep M.; Gonzalez-Plaza, Alejandro; Parrilla, Inmaculada; Rodríguez‐Martínez, H.; Gil, M.A.; Martı́nez, Emilio A.

doi:10.3389/fvets.2021.771996

Cited by 6 publications

(2 citation statements)

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“…As far as we know, this is the first report showing the importance of cryoprotectant equilibration time for vitrification of porcine mature oocytes. Our results clearly demonstrate that the 3-min cryoprotectant equilibration routinely used for porcine morulae and blastocysts (Cuello, Martinez, Cambra, González-Plaza, et al, 2021 was inefficient, achieving viability rates of ~15%. FDA staining is the most consistent method to identify live oocytes without compromising their viability (Shi et al, 2006).…”

Section: Discussionmentioning

confidence: 76%

Equilibration time with cryoprotectants, but not melatonin supplementation during in vitro maturation, affects viability and metaphase plate morphology of vitrified porcine mature oocytes

Gonzalez-Plaza

Brullo²,

Cambra

et al. 2022

Reprod Domestic Animals

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The aims of this study were to investigate the effects of different equilibration times with cryoprotectants on viability and metaphase plate morphology of vitrifiedwarmed porcine mature oocytes (Experiment 1) and to evaluate the effects of supplementation with 10 −9 M melatonin during in vitro maturation on these parameters (Experiment 2). In Experiment 1, 2,392 mature oocytes were vitrified using different equilibration times of oocytes with cryoprotectants (3, 10, 15, 20, 30, 40, 60 and 80 min). Fresh oocytes matured in vitro for 44 hr (n = 509) were used as controls.

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Section: Discussionmentioning

confidence: 76%

Equilibration time with cryoprotectants, but not melatonin supplementation during in vitro maturation, affects viability and metaphase plate morphology of vitrified porcine mature oocytes

Gonzalez-Plaza

Brullo²,

Cambra

et al. 2022

Reprod Domestic Animals

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“…Two facts support this observation. First, morphologically degenerative embryos only presented 47 DEGs compared with normal embryos [24]; second, although vitrification adversely affects pig embryo quality [25,26] and decreases subsequent in vivo embryonic development after ET [27], minor gene expression changes between vitrified and fresh embryos have been reported [28,29].…”

Section: Discussionmentioning

confidence: 99%

The Use of a Brief Synchronization Treatment after Weaning, Combined with Superovulation, Has Moderate Effects on the Gene Expression of Surviving Pig Blastocysts

Gonzalez-Ramiro

Gil

Cuello

et al. 2023

Animals

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The combination of estrus synchronization and superovulation (SS) treatments causes alterations in ovarian and endometrial gene expression patterns, resulting in abnormal follicle and oocyte growth, fertilization, and embryo development. However, the impact of combined SS treatments on the transcriptome of the surviving embryos remains unidentified. In this study, we examined gene expression changes in day 6 blastocysts that survived a brief regimen of synchronization treatment combined with superovulation. The sows were included in one of three groups: SS7 group (n = 6), sows were administered Altrenogest (ALT) 7 days from the day of weaning and superovulated with eCG 24 h after the end of ALT treatment and hCG at the onset of estrus; SO group (n = 6), ALT nontreated sows were superovulated with eCG 24 h postweaning and hCG at the onset of estrus; control group (n = 6), weaned sows displaying natural estrus. Six days after insemination, the sows underwent a surgical intervention for embryo collection. Transcriptome analysis was performed on blastocyst-stage embryos with good morphology. Differentially expressed genes (DEGs) between groups were detected using one-way ANOVA with an un-adjusted p-value < 0.05 and a fold change </> 1.5. The effect of SO treatment on the number of altered pathways and DEGs within each pathway was minimal. Only four pathways were disrupted comprising only a total of four altered transcripts, which were not related to reproductive functions or embryonic development. On the other hand, the surviving blastocysts subjected to SS7 treatments exhibited moderate gene expression changes in terms of DEGs and fold changes, with seven pathways disrupted containing a total of 10 transcripts affected. In this case, the up-regulation of certain pathways, such as the metabolic pathway, with two up-regulated genes associated with reproductive functions, namely RDH10 and SPTLC2, may suggest suboptimal embryo quality, while the down-regulation of others, such as the glutathione metabolism pathway, with down-regulated genes related to cellular detoxification of reactive oxygen species, namely GSTK1 and GSTO1, could depress the embryos’ response to oxidative stress, thereby impairing subsequent embryo development. The gene expression changes observed in the present study in SS7 embryos, along with previous reports indicating SS7 can negatively affect fertilization, embryo production, and reproductive tract gene expression, make its use in embryo transfer programs unrecommendable.

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