Effect of vitrification using the Cryotop method on the gene expression profile of in vitro–produced bovine embryos

Leme, L. O.; Dufort, Isabelle; Sprícigo, J. F. W.; Braga, T. F.; Sirard, Marc‐André; Franco, M. M.; Dode, M. A. N.

doi:10.1016/j.theriogenology.2015.10.016

Cited by 27 publications

(18 citation statements)

References 59 publications

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“…Total RNA was isolated using the RNeasy Plus Micro Kit (Qiagen ® ), according to the manufacturer's instructions. Complementary DNA synthesis was performed as described in deOliveira Leme et al ().…”

Section: Methodsmentioning

confidence: 99%

The dynamics of gene expression, lipid composition and DNA methylation reprogramming are different during in vitro maturation of pig oocytes obtained from prepubertal gilts and cycling sows

Braga

Silva

Marques

et al. 2019

Reprod Domestic Animals

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This study aimed to characterize the gene expression, lipid composition and DNA methylation reprogramming during in vitro maturation (IVM) of pig oocytes with different developmental competencies. We used prepubertal gilts and cycling sows as a model to obtain oocytes with different levels of competency. We found that genes involved in lipid metabolism, SLC27A4, CPT2 and PLIN2, and DNA methylation, DNMT3A, TET1 and TET3, possessed altered transcript expression levels during IVM. Specifically, SLC27A4 mRNA (p = 0.05) increased in oocytes from cycling females, whereas CPT2 (p = 0.05), PLIN2 (p = 0.02) and DNMT3A (p = 0.02) increased in oocytes from prepubertal females during IVM. Additionally, TET3 mRNA increased during IVM in oocytes from prepubertal (p = 0.0005) and cycling females (p = 0.02).The TET1 transcript decreased (p = 0.05) during IVM in oocytes from cycling sows.Regarding lipid composition, mass spectrometry revealed a cluster of ions, with molecular masses higher than m/z 700, which comprises a group of complex phospholipids, was identified in all groups of oocytes, except in those from prepubertal gilts.With respect to DNA methylation reprogramming, it was noted that the less competent oocytes were not able to reprogramme the XIST gene during IVM. We conclude that the maternal mRNA store, lipid composition and epigenetic reprogramming are still being established during maturation and are related to oocyte competence. In addition, we propose that the methylation pattern of the XIST may be used as molecular marker for oocyte competence in pigs. K E Y W O R D S Epigenetics, gene expression, molecular markers, oocyte competence, pig How to cite this article: Braga TF, Silva TCF, Marques MG, et al. The dynamics of gene expression, lipid composition and DNA methylation reprogramming are different during in vitro maturation of pig oocytes obtained from prepubertal gilts and cycling sows.

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Section: Methodsmentioning

confidence: 99%

The dynamics of gene expression, lipid composition and DNA methylation reprogramming are different during in vitro maturation of pig oocytes obtained from prepubertal gilts and cycling sows

Braga

Silva

Marques

et al. 2019

Reprod Domestic Animals

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“…Total RNA was isolated using the RNeasy Plus Micro Kit (Qiagen, Hilden, Germany), according to the manufacturer's instructions. Complementary DNA synthesis was performed as described in Leme et al 62 . The RT-qPCR reactions were performed using the Fast SYBR Green Master Mix Kit (Applied Biosystems, Foster City, California, USA) in an Applied Biosystem 7500 Fast Real Time PCR System (Applied Biosystem).…”

Section: Gene Expression Quantification By Real Time Quantitative Pcrmentioning

confidence: 99%

Intraovarian injection of mesenchymal stem cells improves oocyte yield and in vitro embryo production in a bovine model of fertility loss

Malard

Peixer

Grázia³

et al. 2020

Sci Rep

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Valuable female cattle are continuously subject to follicular puncture (ovum pickup -opU). this technique is commonly used for in-vitro embryo production, but may result in ovarian lesion. Mesenchymal stem cells (MSC) ameliorate the function of injured tissues, but their use to treat ovarian lesions in cattle has not been established. We investigated whether a local injection of MSC would reduce the negative effects of repeated OPU under acute and chronic scenarios in bovines. First, we performed four OPU sessions and injected 2.5 × 10 6 MSCs immediately after the 4th OPU procedure (n = 5). The treated organs (right ovary) were compared to their saline-treated counterparts (left), and presented superior production of oocytes and embryos in the three following OPU sessions (P < 0.05). Then, cows with progressive fertility loss went through three OPU sessions. Animals received MSC, saline, or MSC + FSH in both ovaries after the first OPU. In the two following OPU sessions, the MSC and MSc + FSH-treated groups failed to present any significant alteration in the number of oocytes and embryos compared to saline-treated animals. Thus, MSC have beneficial effects on the fertility of OPUlesioned cows, but not in cows with cystic ovarian disease and chronic ovarian lesions.

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“…We also know that, vitri cation negatively impacts porcine embryo quality, augmenting levels of apoptosis [19,20] or impairing ultrastructure [21]. Vitri cation has been found to cause modi cations in the embryo gene expression in several species [22][23][24][25][26][27]; however, the consequences of vitri cation on gene expression of mammalian embryos are poorly known.…”

Section: Introductionmentioning

confidence: 99%

“…General transcriptome experiments of vitri ed-warmed have been done only in bovine. Vitri cation of in vitro-produced (IVP) bovine embryos caused overexpression of apoptosis-related genes [25] and the repression of genes involved in cell differentiation, cell adhesion and metabolism of lipids [28]. It has been also demonstrated that vitri cation modi ed expression of genes related to stress response [29] in both IVP and in vivo-derived bovine blastocysts.…”

Section: Introductionmentioning

confidence: 99%

Vitrification of in Vivo-derived Porcine Morulae and Blastocysts Alters Metabolic and Stress Response Pathway Alterations

Cuello¹,

Martínez²,

Cambra³

et al. 2020

Preprint

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BackgroundDespite reported promising farrowing rates after non-surgical and surgical transfers of vitrified in vivo-derived porcine morulae and blastocysts (range: 70-75%), their pregnancy loss is yet higher (10-20%) than it is for fresh embryos (< 2.5%). The present study was designed to investigate whether vitrification would affect the transcriptome of porcine morulae and blastocysts, using microarrays and qRT-PCR validation. Material and methods and resultsMorulae and blastocysts were collected from weaned sows (n=13) by laparotomy at Day 6 (Day 0=start of estrus). Of each embryo category, 50 morulae and 50 blastocysts were vitrified (Treatment group) while fresh morulae (n=40) and blastocysts (n=40) acted as control group. After one week of storage, vitrified embryos were cultured in vitrofor 24 h after warming. Non-vitrified morulae (n=40) and blastocysts (n=40), cultured in vitro for 24 h were used as controls. After the in vitro culture period, embryo viability was morphologically assessed. A total of 30 viable embryos per group and embryonic stage(three pools of 10), were subjected to gene expressionanalysis byusing a microarray approach. A fold change cut-off of ±1.5 and a p-unadjusted <0.05 were used to distinguish differentially expressed genes (DEGs). The survival rates of vitrified/warmed morulae and blastocysts were similar to those of the control (nearly 100%, n.s.).A total of 233 DEGs were identified in vitrified morulae (38 upregulated and 195 downregulated) and 105 (112 upregulated and 93 downregulated) in vitrified blastocysts compared to the control group.Transcriptome DEG profiles were dependent on developmental stage, and GO enrichment analysis mainly related DEGs tobiological processes.The vitrification/warming impact on morulae was mostly repression of gene expression with the exception of metabolism-related pathways. In the case of blastocysts, we noted the activation of the cell cycle, cellular senescence andof signaling pathways for TFGβ, p53, FoxO and MAPK. Disruption ofmetabolic-related pathways in morulae and steroid biosynthesis and gap junctions in blastocysts could be related to impaired embryo quality and developmental potential, despite the rather high post-warming survival rates seen in vitro.ConclusionIn conclusion, vitrification modified the transcriptome of in vivo-derived porcine morulae and blastocysts, resulting in moderate gene expression changes, which may disturb subsequent embryo development and pregnancy after embryo transfer.

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Effect of vitrification using the Cryotop method on the gene expression profile of in vitro–produced bovine embryos

Cited by 27 publications

References 59 publications

The dynamics of gene expression, lipid composition and DNA methylation reprogramming are different during in vitro maturation of pig oocytes obtained from prepubertal gilts and cycling sows

The dynamics of gene expression, lipid composition and DNA methylation reprogramming are different during in vitro maturation of pig oocytes obtained from prepubertal gilts and cycling sows

Intraovarian injection of mesenchymal stem cells improves oocyte yield and in vitro embryo production in a bovine model of fertility loss

Vitrification of in Vivo-derived Porcine Morulae and Blastocysts Alters Metabolic and Stress Response Pathway Alterations

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