TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis
Abstract
:1. Introduction
2. Results
2.1. Bobcat339 Treatment Obstructed the First Polar Body Extrusion in Porcine Oocytes
2.2. Bobcat339 Treatment Triggered Apoptosis of Porcine Oocytes
2.3. Effect of Bobcat339 on Spindle Architecture and Chromosomes Alignment in Oocytes
2.4. Bobcat339 Treatment Altered 5mC/5hmC Levels in Porcine Oocytes
2.5. Effect of Bobcat339 on Preimplantation Embryo Development
2.6. Bobcat339 Treatment Decreased the Expression of ZGA and Pluripotency-Related Genes
2.7. Bobcat339 Treatment Disrupted 5mC/5hmC Levels in Porcine Embryos
2.8. Comparative Analyses of Transcriptomic Data
3. Discussion
4. Materials and Methods
4.1. Antibodies and Reagents
4.2. Porcine Oocyte Collection and In Vitro Maturation (IVM)
4.3. Parthenogenetic Activation and In Vitro Culture (IVC)
4.4. Chemical Treatment
4.5. AnnexinV-FITC Staining
4.6. RNA Isolation and RT-PCR
4.7. Immunofluorescent Staining
4.8. RNA Sequencing and Data Analysis
4.9. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Accession | Primer Sequences (5′ to 3′) | Product Size (bp) |
---|---|---|---|
BCL-2 | XM_021077298.1 | F: CAGGGACAGCGTATCAGAGC R: TTGCGATCCGACTCACCAAT | 156 |
BAX | XM_013998624.2 | F: CCAGGATCGAGCAGGGCGAAT R: CACAGGGCCTTGAGCACCAGTTT | 285 |
DPPA2 | XM_003358822.4 | F: CCGTTCCTGCTTCTGTTGAGACC R: GGCGAACCCAACCTTCTGTATCTG | 105 |
EIF1A | NM_001243218.1 | F: GGTGTTCAAAGAAGATGGGCAAGAG R: TTTCCCTCTGATGTGACATAACCTC | 115 |
H19 | AY044827.1 | F: TCAAACGACAAGAGATGGTGCTA R: GACGTCTGTTCCTTTGGCTC | 118 |
NANOG | XM_021092390.1 | F: AGGACAGCCCTGATTCTTCCACAA R: AAAGTTCTTGCATCTGCTGGAGGC | 198 |
OCT4 | XM_021097869.1 | F: AAGCAGTGACTATTCGCAAC R: CAGGGTGGTGAAGTGAGG | 136 |
RPLP0 | NM_001129964.2 | F: GCTAAGGTGCTCGGTTCTTC R: GTGCGGACCAATGCTAGG | 112 |
SOX2 | NM_001123197.1 | F: CGCAGACCTACATGAACG R: TCGGACTTGACCACTGAG | 103 |
TET1 | NM_001315772.1 | F: AGCACAGGACAAAATGAAGG R: TGGTTAGTTGGAGAGGAGG | 171 |
TET2 | XM_013978993.2 | F: GCCAACCCTGTGAACCTCT R: GGGCTGGTAAAGTGTATGG | 270 |
TET3 | XM_021087365.1 | F: TCAAGGCAAAGACCCGAAC R: AGACGGCAGTCAATCGCTATT | 261 |
ZSCAN4 | XM_021097584.1 | F: GCCCAGAAAGTCTTCCCATGTGAG R: GCCTCTCATCATTGTGTCTCCTCTG | 94 |
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Chen, F.; Li, M.-G.; Hua, Z.-D.; Ren, H.-Y.; Gu, H.; Luo, A.-F.; Zhou, C.-F.; Zhu, Z.; Huang, T.; Bi, Y.-Z. TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis. Int. J. Mol. Sci. 2023, 24, 12455. https://doi.org/10.3390/ijms241512455
Chen F, Li M-G, Hua Z-D, Ren H-Y, Gu H, Luo A-F, Zhou C-F, Zhu Z, Huang T, Bi Y-Z. TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis. International Journal of Molecular Sciences. 2023; 24(15):12455. https://doi.org/10.3390/ijms241512455
Chicago/Turabian StyleChen, Fan, Ming-Guo Li, Zai-Dong Hua, Hong-Yan Ren, Hao Gu, An-Feng Luo, Chang-Fan Zhou, Zhe Zhu, Tao Huang, and Yan-Zhen Bi. 2023. "TET Family Members Are Integral to Porcine Oocyte Maturation and Parthenogenetic Pre-Implantation Embryogenesis" International Journal of Molecular Sciences 24, no. 15: 12455. https://doi.org/10.3390/ijms241512455