Vitrification leads to transcriptomic modifications of mice ovaries that do not affect folliculogenesis progression

Pereira, Luiza Aparecida Ansaloni Chagas; Nascimento, Bárbara Rodrigues; Jorge, Érika Cristina; Segatelli, T. M.; Coutinho, Luiz Lehmann; Viana, J. H. M.; Campos-Júnior, Paulo Henrique Almeida

doi:10.1016/j.repbio.2020.02.001

Cited by 9 publications

(6 citation statements)

References 34 publications

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“…Nodes are colored according to the gene abundance (fold change) in comparison pair D10 vs. F; borders are colored according to the gene abundance (fold change) in comparison pair V vs. F. The confidence cutoff was set to 0.4, the confidence score of each interaction is mapped to the edge thickness. Additional file 9 contains the web session of the network for interactive view A recent study using RNA-seq to measure the response of mouse ovarian tissue to vitrification followed by 20 days of auto-transplantation was reported [29]. Additionally, several studies used RNA-seq in recent years to measure global transcriptomic changes in oocytes after vitrification followed by culturing or in vitro maturation [24][25][26][27][28].…”

Section: Discussionmentioning

confidence: 99%

“…Several studies have used RNA sequencing (RNA-seq) to measure global transcriptomic changes in oocytes after vitrification followed by culturing or in vitro maturation [24][25][26][27][28], which provided informative transcriptomic data on the oocyte state after at least several hours post vitrification/ warming. Another recent study performed RNA-seq on mouse ovarian tissue after vitrification followed by 20 days of autotransplantation [29]. It appears essential to investigate cellular transcriptomic changes immediately after completing the preservation/reanimation to track the initial cellular response to potential stresses caused by specific techniques.…”

Section: Introductionmentioning

confidence: 99%

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Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model

Amelkina

Comizzoli

2020

BMC Genomics

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Background Long term preservation of living ovarian tissues is a critical approach in human reproductive medicine as well as in the conservation of rare animal genotypes. Compared to single cell preservation, optimization of protocols for tissues is highly complex because of the diversity of cells responding differently to non-physiological conditions. Using the prepubertal domestic cat as a model, the objective was to study immediate effects of vitrification or microwave-assisted dehydration on the global transcriptome dynamics in the ovarian cortex. RNA sequencing was performed on ovarian tissues (n = 6 individuals) from different conditions: fresh tissue after dissection (F), vitrified/warmed tissue (V), tissue dehydrated for 5 min (D5) or 10 min (D10) followed by rehydration. Differential gene expression analysis was performed for comparison pairs V vs. F, D10 vs. F, D5 vs. F and D10 vs. D5, and networks were built based on results of functional enrichment and in silico protein-protein interactions. Results The impact of the vitrification protocol was already measurable within 20 min after warming and involved upregulation of the expression of seven mitochondrial DNA genes related to mitochondrial respiration. The analysis of D10 vs. F revealed, 30 min after rehydration, major downregulation of gene expression with enrichment of in silico interacting genes in Ras, Rap1, PI3K-Akt and MAPK signaling pathways. However, comparison of D5 vs. F showed negligible effects of the shorter dehydration protocol with two genes enriched in Ras signaling. Comparison of D10 vs. D5 showed downregulation of only seven genes. Vitrification and dehydration protocols mainly changed the expression of different genes and functional terms, but some of the differentially expressed genes formed a major in silico protein-protein interaction cluster enriched for mitochondrial respiration and Ras/MAPK signaling pathways. Conclusions Our results showed, for the first time, different effects of vitrification and microwave-assisted dehydration protocols on the global transcriptome of the ovarian cortex (using the domestic cat as a biomedical model). Acquired data and networks built on the basis of differentially expressed genes (1) can help to better understand stress responses to non-physiological stresses and (2) can be used as directions for future preservation protocol optimizations.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model

Amelkina

Comizzoli

2020

BMC Genomics

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“…Recently, several studies have used RNA-seq to measure the global transcriptomics of oocytes or follicles (Zhang et al, 2020;Jia et al, 2019;Huang et al, 2018;Wang et al, 2017;Liu et al, 2022). Pereira et al, 2020 reported that vitrification and autotransplantation can affect the transcriptomic expression of mouse ovarian tissue. To confirm the safety of vitrified OTC, this study was the first (to the authors' knowledge) to focus on mRNA transcriptomic changes that occur in human ovarian tissues after vitrification.…”

Section: Groupmentioning

confidence: 99%

Impacts of vitrification on the transcriptome of human ovarian tissue in patients with gynecological cancer

Huang

et al. 2023

Front. Genet.

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Objective: This study investigated the effects of a vitrification/warming procedure on the mRNA transcriptome of human ovarian tissues.Design: Human ovarian tissues were collected and processed through vitrification (T-group) and then subjected to RNA sequencing (RNA-seq) analysis, HE, TdT-mediated dUTP nick-end labeling (TUNEL), and real-time quantitative PCR, and the results were compared to those of the fresh group (CK).Results: A total of 12 patients, aged 15–36 years old, with a mean anti-Müllerian hormone level of 4.57 ± 3.31 ng/mL were enrolled in this study. According to the HE and TUNEL results, vitrification effectively preserved human ovarian tissue. A total of 452 significantly dysregulated genes (|log2FoldChange| > 1 and p < 0.05) were identified between the CK and T groups. Among these, 329 were upregulated and 123 were downregulated. A total of 372 genes were highly enriched for 43 pathways (p < 0.05), which were mainly related to systemic lupus erythematous, cytokine–cytokine receptor interaction, the TNF signaling pathway, and the MAPK signaling pathway. IL10, AQP7, CCL2, FSTL3, and IRF7 were significantly upregulated (p < 0.01), while IL1RN, FCGBP, VEGFA, ACTA2, and ASPN were significantly downregulated in the T-group (p < 0.05) compared to the CK group, which agreed with the results of the RNA-seq analysis.Conclusion: These results showed (for the first time to the authors’ knowledge) that vitrification can induce changes in mRNA expression in human ovarian tissues. Further molecular studies on human ovarian tissues are required to determine whether altered gene expression could result in any downstream consequences.

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“…In the third ovarian section, the number of primordial, primary, secondary, antral, and atretic follicles were counted. Follicles were classified as previously described (Campos-Junior et al, 2012a;Pereira et al, 2020) and follicles (from primordial to antral) showing morphological signs of death such as pyknosis, cellular fragmentation, and disintegration, were classified as atretic. Only follicles containing an oocyte with a visible nucleus were considered to avoid double-counting.…”

Section: Histological Evaluations and Follicle Quantificationmentioning

confidence: 99%

“…Values are shown as n-fold difference. All procedures were performed as previously described by our group (Campos-Junior et al, 2016;Pereira et al, 2020).…”

Section: Qrt-pcrmentioning

confidence: 99%

The CC-chemokine receptor 2 is involved in the control of ovarian folliculogenesis and fertility lifespan in mice

Santos

Pereira

Viana

et al. 2020

Journal of Reproductive Immunology

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Vitrification leads to transcriptomic modifications of mice ovaries that do not affect folliculogenesis progression

Cited by 9 publications

References 34 publications

Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model

Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model

Impacts of vitrification on the transcriptome of human ovarian tissue in patients with gynecological cancer

The CC-chemokine receptor 2 is involved in the control of ovarian folliculogenesis and fertility lifespan in mice

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