ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos
Abstract
:1. Introduction
2. Results
2.1. Expression and Localization of ZSCAN4 in Porcine Embryos
2.2. Effects of ZSCAN4 KD on Porcine Embryonic Development
2.3. Effect of ZSCAN4 KD on Histone Modifications and ZGA
2.4. ZSCAN4 KD Induced Global DNA Methylation
2.5. ZSCAN4 Regulated DNMT1 Expression to Stabilize Telomere Length
2.6. ZSCAN4 KD Induced DNA Damage and Apoptosis in Porcine Embryos
3. Materials and Method
3.1. ZSCAN4 dsRNA and DNMT1 dsRNA Preparation
3.2. Oocyte Harvest and In Vitro Maturation
3.3. Parthenogenetic Activation, In Vitro Culture, and dsRNA Injection
3.4. Quantitative Reverse Transcription PCR (qRT-PCR)
3.5. Immunofluorescence and Confocal Microscopy
3.6. Western Blot Analysis
3.7. Statistical Analysis
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Zhou, W.; Niu, Y.J.; Nie, Z.W.; Kim, J.Y.; Xu, Y.N.; Yan, C.G.; Cui, X.S. Nuclear accumulation of pyruvate dehydrogenase alpha 1 promotes histone acetylation and is essential for zygotic genome activation in porcine embryos. Biochim. Biophys. Acta Mol. Cell Res. 2020, 1867, 118648. [Google Scholar] [CrossRef]
- Magnani, L.; Johnson, C.M.; Cabot, R.A. Expression of eukaryotic elongation initiation factor 1A differentially marks zygotic genome activation in biparental and parthenogenetic porcine embryos and correlates with in vitro developmental potential. Reprod. Fertil. Dev. 2008, 20, 818–825. [Google Scholar] [CrossRef] [PubMed]
- De Sousa, P.A.; Watson, A.J.; Schultz, R.M. Transient expression of a translation initiation factor is conservatively associated with embryonic gene activation in murine and bovine embryos. Biol. Reprod. 1998, 59, 969–977. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yan, L.; Yang, M.; Guo, H.; Yang, L.; Wu, J.; Li, R.; Liu, P.; Lian, Y.; Zheng, X.; Yan, J.; et al. Single-cell RNA-Seq profiling of human preimplantation embryos and embryonic stem cells. Nat. Struct. Mol. Biol. 2013, 20, 1131–1139. [Google Scholar] [CrossRef]
- Lee, M.T.; Bonneau, A.R.; Takacs, C.M.; Bazzini, A.A.; DiVito, K.R.; Fleming, E.S.; Giraldez, A.J. Nanog, Pou5f1 and SoxB1 activate zygotic gene expression during the maternal-to-zygotic transition. Nature 2013, 503, 360–364. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xue, L.; Cai, J.Y.; Ma, J.; Huang, Z.; Guo, M.X.; Fu, L.Z.; Shi, Y.B.; Li, W.X. Global expression profiling reveals genetic programs underlying the developmental divergence between mouse and human embryogenesis. BMC Genom. 2013, 14, 568. [Google Scholar] [CrossRef] [Green Version]
- Zhou, W.; Nie, Z.W.; Zhou, D.J.; Cui, X.S. Acetyl-CoA synthases are essential for maintaining histone acetylation under metabolic stress during zygotic genome activation in pigs. J. Cell. Physiol. 2021, 236, 6948–6962. [Google Scholar] [CrossRef]
- Falco, G.; Lee, S.L.; Stanghellini, I.; Bassey, U.C.; Hamatani, T.; Ko, M.S. ZSCAN4: A novel gene expressed exclusively in late 2-cell embryos and embryonic stem cells. Dev. Biol. 2007, 307, 539–550. [Google Scholar] [CrossRef] [Green Version]
- Zalzman, M.; Falco, G.; Sharova, L.V.; Nishiyama, A.; Thomas, M.; Lee, S.L.; Stagg, C.A.; Hoang, H.G.; Yang, H.T.; Indig, F.E.; et al. ZSCAN4 regulates telomere elongation and genomic stability in ES cells. Nature 2010, 464, 858–863. [Google Scholar] [CrossRef] [Green Version]
- Akiyama, T.; Xin, L.; Oda, M.; Sharov, A.A.; Amano, M.; Piao, Y.; Cadet, J.S.; Dudekula, D.B.; Qian, Y.; Wang, W.; et al. Transient bursts of ZSCAN4 expression are accompanied by the rapid derepression of heterochromatin in mouse embryonic stem cells. DNA Res. 2015, 22, 307–318. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Amano, T.; Hirata, T.; Falco, G.; Monti, M.; Sharova, L.V.; Amano, M.; Sheer, S.; Hoang, H.G.; Piao, Y.; Stagg, C.A.; et al. ZSCAN4 restores the developmental potency of embryonic stem cells. Nat. Commun. 2013, 4, 1966. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dan, J.; Li, M.; Yang, J.; Li, J.; Okuka, M.; Ye, X.; Liu, L. Roles for Tbx3 in regulation of two-cell state and telomere elongation in mouse ES cells. Sci. Rep. 2013, 3, 3492. [Google Scholar] [CrossRef] [Green Version]
- Wang, Z.X.; Teh, C.H.; Kueh, J.L.; Lufkin, T.; Robson, P.; Stanton, L.W. Oct4 and Sox2 directly regulate expression of another pluripotency transcription factor, Zfp206, in embryonic stem cells. J. Biol. Chem. 2007, 282, 12822–12830. [Google Scholar] [CrossRef] [Green Version]
- Takahashi, K.; Ross, P.J.; Sawai, K. The necessity of ZSCAN4 for preimplantation development and gene expression of bovine embryos. J. Reprod. Dev. 2019, 65, 319–326. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hirata, T.; Amano, T.; Nakatake, Y.; Amano, M.; Piao, Y.; Hoang, H.G.; Ko, M.S. ZSCAN4 transiently reactivates early embryonic genes during the generation of induced pluripotent stem cells. Sci. Rep. 2012, 2, 208. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cheng, Z.L.; Zhang, M.L.; Lin, H.P.; Gao, C.; Song, J.B.; Zheng, Z.; Li, L.; Zhang, Y.; Shen, X.; Zhang, H.; et al. The ZSCAN4-Tet2 Transcription Nexus Regulates Metabolic Rewiring and Enhances Proteostasis to Promote Reprogramming. Cell Rep. 2020, 32, 107877. [Google Scholar] [CrossRef]
- Portney, B.A.; Arad, M.; Gupta, A.; Brown, R.A.; Khatri, R.; Lin, P.N.; Hebert, A.M.; Angster, K.H.; Silipino, L.E.; Meltzer, W.A.; et al. ZSCAN4 facilitates chromatin remodeling and promotes the cancer stem cell phenotype. Oncogene 2020, 39, 4970–4982. [Google Scholar] [CrossRef]
- Le, R.; Huang, Y.; Zhang, Y.; Wang, H.; Lin, J.; Dong, Y.; Li, Z.; Guo, M.; Kou, X.; Zhao, Y.; et al. Dcaf11 activates ZSCAN4-mediated alternative telomere lengthening in early embryos and embryonic stem cells. Cell Stem Cell 2021, 28, 732–747.e739. [Google Scholar] [CrossRef]
- Blackburn, E.H. Switching and signaling at the telomere. Cell 2001, 106, 661–673. [Google Scholar] [CrossRef] [Green Version]
- Hiyama, E.; Hiyama, K. Telomere and telomerase in stem cells. Br. J. Cancer 2007, 96, 1020–1024. [Google Scholar] [CrossRef] [Green Version]
- Blasco, M.A. Telomeres and human disease: Ageing, cancer and beyond. Nat. Rev. Genet. 2005, 6, 611–622. [Google Scholar] [CrossRef] [PubMed]
- Dan, J.; Zhou, Z.; Wang, F.; Wang, H.; Guo, R.; Keefe, D.L.; Liu, L. ZSCAN4 Contributes to Telomere Maintenance in Telomerase-Deficient Late Generation Mouse ESCs and Human ALT Cancer Cells. Cells 2022, 11, 456. [Google Scholar] [CrossRef] [PubMed]
- Zvereva, M.I.; Shcherbakova, D.M.; Dontsova, O.A. Telomerase: Structure, functions, and activity regulation. Biochemistry 2010, 75, 1563–1583. [Google Scholar] [CrossRef] [PubMed]
- Wright, W.E.; Piatyszek, M.A.; Rainey, W.E.; Byrd, W.; Shay, J.W. Telomerase activity in human germline and embryonic tissues and cells. Dev. Genet. 1996, 18, 173–179. [Google Scholar] [CrossRef]
- Liu, L.; Bailey, S.M.; Okuka, M.; Munoz, P.; Li, C.; Zhou, L.; Wu, C.; Czerwiec, E.; Sandler, L.; Seyfang, A.; et al. Telomere lengthening early in development. Nat. Cell Biol. 2007, 9, 1436–1441. [Google Scholar] [CrossRef]
- Dan, J.; Rousseau, P.; Hardikar, S.; Veland, N.; Wong, J.; Autexier, C.; Chen, T. ZSCAN4 Inhibits Maintenance DNA Methylation to Facilitate Telomere Elongation in Mouse Embryonic Stem Cells. Cell Rep. 2017, 20, 1936–1949. [Google Scholar] [CrossRef] [Green Version]
- Van Thuan, N.; Harayama, H.; Miyake, M. Characteristics of preimplantational development of porcine parthenogenetic diploids relative to the existence of amino acids in vitro. Biol. Reprod. 2002, 67, 1688–1698. [Google Scholar] [CrossRef] [Green Version]
- Cui, X.S.; Jeong, Y.J.; Lee, H.Y.; Cheon, S.H.; Kim, N.H. Fetal bovine serum influences apoptosis and apoptosis-related gene expression in porcine parthenotes developing in vitro. Reproduction 2004, 127, 125–130. [Google Scholar] [CrossRef] [Green Version]
- Zhang, J.M.; Genois, M.M.; Ouyang, J.; Lan, L.; Zou, L. Alternative lengthening of telomeres is a self-perpetuating process in ALT-associated PML bodies. Mol. Cell 2021, 81, 1027–1042.e1024. [Google Scholar] [CrossRef]
- Roos, W.P.; Kaina, B. DNA damage-induced cell death by apoptosis. Trends Mol. Med. 2006, 12, 440–450. [Google Scholar] [CrossRef] [PubMed]
- Meulmeester, E.; Jochemsen, A.G. p53: A guide to apoptosis. Curr. Cancer Drug Targets 2008, 8, 87–97. [Google Scholar] [CrossRef] [PubMed]
- Zhou, D.; Niu, Y.; Cui, X.-S. M-RAS Regulate CDH1 Function in Blastomere Compaction during Porcine Embryonic Development. J. Anim. Reprod. Biotechnol. 2020, 35, 12–20. [Google Scholar] [CrossRef]
- Zhou, D.; Li, X.-H.; Lee, S.-H.; Heo, G.; Cui, X.-S. Effects of alpha-linolenic acid and essential amino acids on the proliferation and differentiation of C2C12 myoblasts. J. Anim. Reprod. Biotechnol. 2022, 37, 17–26. [Google Scholar] [CrossRef]
- Shaw, L.; Sneddon, S.F.; Brison, D.R.; Kimber, S.J. Comparison of gene expression in fresh and frozen-thawed human preimplantation embryos. Reproduction 2012, 144, 569–582. [Google Scholar] [CrossRef] [Green Version]
- Vassena, R.; Boue, S.; Gonzalez-Roca, E.; Aran, B.; Auer, H.; Veiga, A.; Izpisua Belmonte, J.C. Waves of early transcriptional activation and pluripotency program initiation during human preimplantation development. Development 2011, 138, 3699–3709. [Google Scholar] [CrossRef] [Green Version]
- Kigami, D.; Minami, N.; Takayama, H.; Imai, H. MuERV-L is one of the earliest transcribed genes in mouse one-cell embryos. Biol. Reprod. 2003, 68, 651–654. [Google Scholar] [CrossRef] [Green Version]
- Probst, A.V.; Okamoto, I.; Casanova, M.; El Marjou, F.; Le Baccon, P.; Almouzni, G. A strand-specific burst in transcription of pericentric satellites is required for chromocenter formation and early mouse development. Dev. Cell 2010, 19, 625–638. [Google Scholar] [CrossRef] [Green Version]
- Karlic, R.; Chung, H.R.; Lasserre, J.; Vlahovicek, K.; Vingron, M. Histone modification levels are predictive for gene expression. Proc. Natl. Acad. Sci. USA 2010, 107, 2926–2931. [Google Scholar] [CrossRef]
- Sun, M.H.; Jiang, W.J.; Li, X.H.; Lee, S.H.; Heo, G.; Zhou, D.; Choi, J.S.; Kim, K.S.; Lv, W.; Cui, X.S. ATF7-dependent epigenetic changes induced by high temperature during early porcine embryonic development. Cell Prolif. 2023, 56, e13352. [Google Scholar] [CrossRef]
- Newell-Price, J.; Clark, A.J.; King, P. DNA methylation and silencing of gene expression. Trends Endocrinol. Metab. 2000, 11, 142–148. [Google Scholar] [CrossRef]
- Deng, M.; Zhang, G.; Cai, Y.; Liu, Z.; Zhang, Y.; Meng, F.; Wang, F.; Wan, Y. DNA methylation dynamics during zygotic genome activation in goat. Theriogenology 2020, 156, 144–154. [Google Scholar] [CrossRef] [PubMed]
- Eckersley-Maslin, M.A.; Svensson, V.; Krueger, C.; Stubbs, T.M.; Giehr, P.; Krueger, F.; Miragaia, R.J.; Kyriakopoulos, C.; Berrens, R.V.; Milagre, I.; et al. MERVL/ZSCAN4 Network Activation Results in Transient Genome-wide DNA Demethylation of mESCs. Cell Rep. 2016, 17, 179–192. [Google Scholar] [CrossRef] [Green Version]
- Ozturk, S.; Sozen, B.; Demir, N. Telomere length and telomerase activity during oocyte maturation and early embryo development in mammalian species. Mol. Hum. Reprod. 2014, 20, 15–30. [Google Scholar] [CrossRef] [Green Version]
- Zhao, Y.; Hoshiyama, H.; Shay, J.W.; Wright, W.E. Quantitative telomeric overhang determination using a double-strand specific nuclease. Nucleic Acids Res. 2008, 36, e14. [Google Scholar] [CrossRef] [PubMed]
- Frias, C.; Pampalona, J.; Genesca, A.; Tusell, L. Telomere dysfunction and genome instability. Front. Biosci. 2012, 17, 2181–2196. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shay, J.W.; Wright, W.E. Senescence and immortalization: Role of telomeres and telomerase. Carcinogenesis 2005, 26, 867–874. [Google Scholar] [CrossRef] [PubMed]
- Ko, M.S. Zygotic Genome Activation Revisited: Looking through the Expression and Function of ZSCAN4. Curr. Top. Dev. Biol. 2016, 120, 103–124. [Google Scholar] [CrossRef]
- Jiang, J.; Lv, W.; Ye, X.; Wang, L.; Zhang, M.; Yang, H.; Okuka, M.; Zhou, C.; Zhang, X.; Liu, L.; et al. ZSCAN4 promotes genomic stability during reprogramming and dramatically improves the quality of iPS cells as demonstrated by tetraploid complementation. Cell Res. 2013, 23, 92–106. [Google Scholar] [CrossRef] [Green Version]
- Srinivasan, R.; Nady, N.; Arora, N.; Hsieh, L.J.; Swigut, T.; Narlikar, G.J.; Wossidlo, M.; Wysocka, J. ZSCAN4 binds nucleosomal microsatellite DNA and protects mouse two-cell embryos from DNA damage. Sci. Adv. 2020, 6, eaaz9115. [Google Scholar] [CrossRef] [Green Version]
Gene | Forward Primer | Reverse Primer |
---|---|---|
ds-ZSCAN4 | 5′-GCCCTCTTTTCTGAGAATATGCC-3′ | 5′-CTGATGGACTTTCAACCGAGA-3′ |
ds-DNMT1 | 5′-CAAACTACCAGGCAGACCAC-3′ | 5′-CACTACTGCCGTTTTGGTTCG-3′ |
ZSCAN4 | 5′-CTTGTTTGGTCCTCGAACAGT-3′ | 5′-TTCATGCCATCGTCTGTCAGGT-3′ |
DNMT1 | 5′-CTCCCTACAGAAGAACCGGAA-3′ | 5′-CCTCGTGCTTCTGTCTAGCTC-3′ |
Telomere | 5′-GGT TTT TGA GGG TGA GGG TGA GGG TGA GGG TGA GGG T-3′ | 5′-TCC CGA CTA TCC CTA TCC CTA TCC CTA TCC CTA TCC CTA-3′ |
36B4 | 5′-TGAAGTGCTTGACATCACCGAGGA-3′ | 5′-CTGCAGACATACGCTGGCAACATT-3′ |
Caspase3 | 5′-TCTAACTGGCAAACCCAAACTT-3′ | 5′-AGTCCCACTGTCCGTCTCAAT-3′ |
BAX | 5′-GAATGGGGGGAGAGACACCT-3′ | 5′-CCGCCACTCGGAAAAAGA-3′ |
BCL2 | 5′-GAACTGGGGGAGGATTGTGG-3′ | 5′-CATCCCAGCCTCCGTTATCC-3′ |
18s | 5′-CGCGGTTCTATTTTGTTGGT’-3′ | 5′-AGTCGGCATCGTTTATGGTC-3′ |
TCSTV3 | 5′-AGAAAGGGCTGGAACTTGTGACCT-3′ | 5′-AAAGCTCTTTGAAGCCATGCCCAG-3′ |
WEE1 | 5′-ACCTCGGATTCCACAAGTGCTTT-3′ | 5′-ATGCTTTACCAGTGCCATTGCT-3′ |
RIF1 | 5′-TTGACATCATTTTACCGCAGA-3′ | 5′-GGAATCTCTTCTAGTCCACGA-3′ |
TBX3 | 5′-CTGACCCCGAAATGCCGAAG-3′ | 5′-ACTATAATTCCCCTGCCACGTT-3′ |
EIF1A | 5′-GGTGTTCAAAGAAGATGGGCAAGAG-3′ | 5′-TTTCCCTCTGATGTGACATAACCTC-3′ |
DPPA2 | 5′-TACAGAAGGTTGGGTTCGCC-3′ | 5′-GGTCTGGGGATGGGAAAGTG-3′ |
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Li, X.-H.; Sun, M.-H.; Jiang, W.-J.; Zhou, D.; Lee, S.-H.; Heo, G.; Chen, Z.; Cui, X.-S. ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos. Int. J. Mol. Sci. 2023, 24, 12121. https://doi.org/10.3390/ijms241512121
Li X-H, Sun M-H, Jiang W-J, Zhou D, Lee S-H, Heo G, Chen Z, Cui X-S. ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos. International Journal of Molecular Sciences. 2023; 24(15):12121. https://doi.org/10.3390/ijms241512121
Chicago/Turabian StyleLi, Xiao-Han, Ming-Hong Sun, Wen-Jie Jiang, Dongjie Zhou, Song-Hee Lee, Geun Heo, Zhi Chen, and Xiang-Shun Cui. 2023. "ZSCAN4 Regulates Zygotic Genome Activation and Telomere Elongation in Porcine Parthenogenetic Embryos" International Journal of Molecular Sciences 24, no. 15: 12121. https://doi.org/10.3390/ijms241512121