Table 4.
Impact of oocyte vitrification on expression in animals
| References | Animal Model | Materials | Number of oocytes and embryos | Technology of assessment | Studied sequences | Conclusions |
|---|---|---|---|---|---|---|
| Wu et al. [129] | Murine | MI and MII after IVM from fresh and vitrified GV | 20–25 per group | RTqPCR | Mps1, BubR1, Mad1, Mad2 | Expression of spindle assembly checkpoint (SAC)-related genes in GV (Mad1, BubR1, and Mad2), and MII stages (Mps1 and Mad1) were significantly downregulated after vitrification. |
| Chen et al. [130] | Murine | fresh and vitrified MII | 50 per group | RTqPCR | Gtl2, Peg10, Sirt1, Peg3, Igf2R, H19, Igf2 | Gtl2 and Peg10 were significantly increased, Peg3, Igf2R, and Sirt1 were significantly decreased after vitrification |
| Jia et al. [127] | Porcine | MII after IVM from vitrified and fresh COC | 25 per group |
RNAseq RTqPCR validation |
Global analysis (Illumina) 21 genes |
Significant differential expression between the non-cryopreserved and vitrified oocyte pools (19 upregulated genes and 18 downregulated after vitrification and IVM). No GO enrichment or KEGG pathway was identified. |
| Huang et al. [126] | Bovine | GV, MII after IVM from vitrified GV | 3 fresh GV, 4 vitrified GV, 1 fresh MII, and 2 MII derived from vitrified GV | RNAseq | Global analysis (Illumina) |
For GV, 12 upregulated genes and 19 downregulated genes after vitrification. No GO enrichment or KEGG pathway was identified. For MII, 47 upregulated genes and 6 downregulated genes after vitrification. With GO and KEGG analyses, several pathways were identified: transcription regulation, cell differentiation and mitosis, regulation of actin cytoskeleton, and apoptosis. |
| Ma et al. [131] | Bovine | MII after IVM from fresh GV, MII after IVM from vitrified GV, fresh GV, vitrified GV | 15 per group (*3 experiments) | RTqPCR | CD9, CD81, DNMT1, and DNMT3b | The expression of all analysed genes was downregulated after IVM of vitrified GV when compared to the fresh in vitro matured MII oocytes. |
| Gao et al. [125] | Murine | MII after IVM from fresh GV, MII after IVM from vitrified GV, fresh MII, vitrified MII | 100 per group |
RNAseq RTqPCR validation |
Global analysis (Illumina) Atp5e, Atp5o, Ndufb9, Uqcrq, Timm17a, Dppa5a, H3f3a, Timm13, and Tomm40 |
No effect of vitrification on the transcriptome. Differences were reported for IVM. |
| Wu et al. [132] | Bovine | MII after IVM from vitrified GV (liquid nitrogen-LN or helium-LHe), MII after IVM from fresh GV | 120 per group | RTqPCR | p53, EG5, CDC20, and NPM2 |
For LN-effet, p53 and EG5 were upregulated after vitrification, and CDC20 was downregulated. For LHe-effect, lower effect on the expression of some related genes compared to LN vitrification. |
| Wang et al. [128] | Bovine | MII after IVM from vitrified GV, MII after IVM from fresh GV | 20 per group |
RNAseq RTqPCR validation |
Global analysis (Illumina) CDK2,UCHL3, CALM, VDAC2, DPH6, MED27, DAD1, MED21, NR1H4, and HMGN1 | Significant differential expression between the non-cryopreserved and vitrified oocyte pools (12 upregulated genes and 90 downregulated genes). At GO analysis, several enrichments in terms of membrane–bounded organelles, macromolecular complex, and intra-cellular part were found. No KEGG pathway was identified. |
| Shirazi et al. [133] | Ovine | Fresh MII, vitrified MII, fresh GV, vitrified GV | 25 per group | RTqPCR | STAT3, HAT1, HDAC1, SUV39H1, DNMT1, HMGN3a, SMARCAL1, and DNMT3b | The HMG3a and HDAC1 expression was downregulated after vitrification. |
| Zhao et al. [27] | Bovine | MII after IVM from vitrified GV, MII after IVM from fresh GV | 100 per group | RTqPCR | BAX and BCL2 l1 | The BAX expression was upregulated and the BCL2 l1 expression was downregulated after vitrification. |
| Dai et al. [134] | Porcine | Fresh MII, vitrified MII | 100 per group | RTqPCR | DNM1, SOD1, MFN2, BAX, and BCL2 | The DNM1 expression was upregulated, and the SOD1, MFN2, BAX, and BCL2 were downregulated after vitrification. |
| Spricigo et al. [135] | Bovine | MII after IVM from vitrified COC, MII after IVM from fresh COC | 20 per group (*4 experiments) | RTqPCR | DNMT1, SUV39H1, HDAC2, TP53, and CASP3 | No effect of vitrification. |
| Cheng et al. [113] | Murine | Fresh MII, vitrified MII | 100 per group | RTqPCR | Dnmt1,3a,3b,3 l | The DNMTs expression was significantly reduced after vitrification. |
| Zhao et al. [114] | Murine | Fresh MII, vitrified MII | 200 per group | RTqPCR | Dnmt1o, Hat1, and Hdac1 promoters |
No effect of vitrification on the mRNA expression levels of Hat1 and Hdac1. The Dnmt1o expression was significantly reduced after vitrification. |
| Zhou et al. [136] | Bovine | Fresh MII, vitrified MII | 50 per group | RTqPCR | CD9 | The CD9 expression was downregulated after vitrification. |
| Rao et al. [137] | Caprine | MII after IVM from vitrified and fresh COC | 60 per group | RTqPCR | GDF9, BMP15, TGFBR1, BPR2, BCL2, BAX and P53 | No significant differences of most of the genes. |
| Turathum et al. [138] | Canine | MII after IVM from vitrified and fresh COC | 200 vitrified, 292 fresh | RTqPCR | HSP70, Dnmt1, SOD1, BAX, and Bcl2 | Bcl2 expression was increased after vitrification, whereas BAX was not expressed in both groups. |
| Anchamparuthy et al. [139] | Bovine | Fresh GV, MII after IVM from vitrified GV, MII after IVM from fresh GV | 25 per group | RT-qPCR | 18S rRNA, Fas, FasL, Bax, and Bcl-2 | The Bax expression was upregulated after vitrification with the ratio of Bax:Bcl-2 elevated. |
| Habibi et al. [140] | Murine | MII after IVM from vitrified GV, MII after IVM from fresh GV | 10 per group (*3 experiments) | RT-qPCR | Mater, Sod1, and Hook1 | The Mater and Hook1 expression was downregulated after vitrification. |
| Succu et al. [141] | Ovine | MII after IVM from vitrified GV, MII after IVM from fresh GV | 40 vitrified, 24 fresh | RT-qPCR | b-actin, H2A.Z histone, Poli A, PAP, HSP90b, P34cdc2, Cyclin B, Na/K-ATPase and Type I cadherin | Except for the b-actin and H2A.Z expression, all gene expression was downregulated after vitrification. |
GV oocyte at germinal vesicle stage, IVM in vitro maturation, KEGG Kyoto Encyclopedia of Genes and Genomes, LN liquid nitrogen; LHe = liquid helium, MII oocyte at metaphase II stage, GO gene ontology, RNAseq RNA sequencing, RT-qPCR quantitative reverse transcription PCR