{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T13:30:09Z","timestamp":1775136609222,"version":"3.50.1"},"reference-count":53,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2022,2,28]],"date-time":"2022-02-28T00:00:00Z","timestamp":1646006400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31802059"],"award-info":[{"award-number":["31802059"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31902226"],"award-info":[{"award-number":["31902226"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003995","name":"Anhui Provincial Natural Science Foundation","doi-asserted-by":"publisher","award":["2008085MC85"],"award-info":[{"award-number":["2008085MC85"]}],"id":[{"id":"10.13039\/501100003995","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Zygotic genome activation (ZGA) plays an essential role in early embryonic development. Vitrification is a common assisted reproductive technology that frequently reduces the developmental competence of embryos. However, the effect of vitrification on porcine ZGA and gene expression during ZGA remains largely unclear. Here, we found that vitrification of pronuclear zygotes derived from parthenogenetic activation (PA) and in vitro fertilization (IVF) resulted in a significant reduction in the rates of 2-cell, 4-cell, and blastocysts, but did not affect the quality of blastocysts. Functional research revealed that RNA polymerase II Inhibitor (\u03b1-amanitin) treatment significantly reduced global transcriptional activity and developmental efficiency of both 4-cell and 8-cell embryos, implying an essential role of ZGA in porcine early embryonic development. Furthermore, vitrification did not affect the synthesis of nascent mRNA of 2-cell embryos, but significantly inhibited global transcriptional activity of both 4-cell and 8-cell embryos, suggesting an impaired effect of vitrification on porcine ZGA. Correspondingly, the single-cell analysis showed that vitrification caused the downregulation or upregulation expression of maternal genes in 4-cell embryos, also significantly decreased the expression of zygotic genes. Taken together, these results indicated that vitrification of pronuclear zygotes impairs porcine zygotic genome activation.<\/jats:p>","DOI":"10.3390\/ani12050610","type":"journal-article","created":{"date-parts":[[2022,2,28]],"date-time":"2022-02-28T20:09:57Z","timestamp":1646078997000},"page":"610","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Vitrification of Pronuclear Zygotes Perturbs Porcine Zygotic Genome Activation"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1372-9389","authenticated-orcid":false,"given":"Tengteng","family":"Xu","sequence":"first","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chengxue","family":"Liu","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mengya","family":"Zhang","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin","family":"Wang","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yelian","family":"Yan","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qiuchen","family":"Liu","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yangyang","family":"Ma","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tong","family":"Yu","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anucha","family":"Sathanawongs","sequence":"additional","affiliation":[{"name":"Department of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Jiao","sequence":"additional","affiliation":[{"name":"Anhui Haoyu Animal Husbandry Co., Ltd., Luan 237451, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8916-8154","authenticated-orcid":false,"given":"Zubing","family":"Cao","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9721-5910","authenticated-orcid":false,"given":"Yunhai","family":"Zhang","sequence":"additional","affiliation":[{"name":"Anhui Province Key Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.anireprosci.2013.02.006","article-title":"Cryobanking of farm animal gametes and embryos as a means of conserving livestock genetics","volume":"138","author":"Mara","year":"2013","journal-title":"Anim. Reprod. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1637","DOI":"10.1016\/j.theriogenology.2016.07.018","article-title":"Cryopreservation of animal oocytes and embryos: Current progress and future prospects","volume":"86","author":"Mandawala","year":"2016","journal-title":"Theriogenology"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1007\/s10815-009-9356-0","article-title":"Vitrification of human early cavitating and deflated expanded blastocysts: Clinical outcome of 474 cycles","volume":"26","author":"Raju","year":"2009","journal-title":"J. Assist. Reprod. Gen."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1186\/1477-7827-7-99","article-title":"Factors affecting the outcome of human blastocyst vitrification","volume":"7","author":"Kader","year":"2009","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1530\/REP-16-0480","article-title":"Effect of vitrification on in vitro development and imprinted gene Grb10 in mouse embryos","volume":"154","author":"Yao","year":"2017","journal-title":"Reproduction"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1111\/asj.12892","article-title":"Vitrification of bovine matured oocytes and blastocysts in a paper container","volume":"89","author":"Paul","year":"2018","journal-title":"Anim. Sci. J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.cryobiol.2017.08.002","article-title":"Embryo survival and birth rate after minimum volume vitrification or slow freezing of in vivo and in vitro produced ovine embryos","volume":"78","author":"Cuadro","year":"2017","journal-title":"Cryobiology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1095\/biolreprod.108.070235","article-title":"Live Piglets Derived from In Vitro-Produced Zygotes Vitrified at the Pronuclear Stage","volume":"80","author":"Somfai","year":"2009","journal-title":"Biol. Reprod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1095\/biolreprod.108.073387","article-title":"Production of Piglets after Cryopreservation of Embryos Using a Centrifugation-Based Method for Delipation Without Micromanipulation1","volume":"80","author":"Li","year":"2009","journal-title":"Biol. Reprod."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1002\/mrd.10282","article-title":"Effects of cooling on meiotic spindle structure and chromosome alignment within in vitro matured porcine oocytes","volume":"65","author":"Liu","year":"2003","journal-title":"Mol. Reprod. Dev."},{"key":"ref_11","first-page":"1159","article-title":"Vitrification of in vitro matured oocytes: Effects on meiotic spindle configuration and mitochondrial function","volume":"7","author":"Lei","year":"2014","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.cryobiol.2015.08.002","article-title":"Changes in mitochondrial function in porcine vitrified MII-stage oocytes and their impacts on apoptosis and developmental ability","volume":"71","author":"Dai","year":"2015","journal-title":"Cryobiology"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Hara, T., Kin, A., Aoki, S., Nakamura, S., Shirasuna, K., Kuwayama, T., and Iwata, H. (2018). Resveratrol enhances the clearance of mitochondrial damage by vitrification and improves the development of vitrified-warmed bovine embryos. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0204571"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1002\/mrd.23081","article-title":"BMP4 plays a role in apoptosis during human preimplantation development","volume":"86","author":"Aberkane","year":"2019","journal-title":"Mol. Reprod. Dev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1016\/j.theriogenology.2016.03.008","article-title":"Effects of oocyte vitrification on epigenetic status in early bovine embryos","volume":"86","author":"Chen","year":"2016","journal-title":"Theriogenology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.cryobiol.2017.12.001","article-title":"Vitrification, not cryoprotectant exposure, alters the expression of developmentally important genes in in vitro produced porcine blastocysts","volume":"80","author":"Bartolac","year":"2018","journal-title":"Cryobiology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1002\/wdev.220","article-title":"The mammalian blastocyst","volume":"5","author":"Frankenberg","year":"2016","journal-title":"Wires Dev. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1111\/boc.201700029","article-title":"Mechanics of blastocyst morphogenesis","volume":"109","year":"2017","journal-title":"Biol. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1002\/j.1460-2075.1982.tb01230.x","article-title":"The transition from maternal to embryonic control in the 2-cell mouse embryo","volume":"1","author":"Flach","year":"1982","journal-title":"EMBO J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"118648","DOI":"10.1016\/j.bbamcr.2020.118648","article-title":"Nuclear accumulation of pyruvate dehydrogenase alpha 1 promotes histone acetylation and is essential for zygotic genome activation in porcine embryos","volume":"1867","author":"Zhou","year":"2020","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1530\/jrf.0.0860637","article-title":"Qualitative and quantitative changes in protein synthesis occur at the 8-16-cell stage of embryogenesis in the cow","volume":"86","author":"Frei","year":"1989","journal-title":"J. Reprod. Fertil."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1530\/jrf.0.0820769","article-title":"Control of protein synthesis during early cleavage of sheep embryos","volume":"82","author":"Crosby","year":"1988","journal-title":"J. Reprod. Fertil."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1038\/332459a0","article-title":"Human gene expression first occurs between the four- and eight-cell stages of preimplantation development","volume":"332","author":"Braude","year":"1988","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1530\/jrf.0.0500147","article-title":"Effects of -amanitin on the development of mouse ova in culture","volume":"50","author":"Levey","year":"1977","journal-title":"J. Reprod. Fertil."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1038\/248678a0","article-title":"In vivo and in vitro effect of alpha-amanitin on preimplantation mouse embryo RNA polymerase","volume":"248","author":"Warner","year":"1974","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1016\/j.cub.2018.03.058","article-title":"Depletion of Maternal Cyclin B3 Contributes to Zygotic Genome Activation in the Ciona Embryo","volume":"28","author":"Treen","year":"2018","journal-title":"Curr. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1093\/biolre\/ioz012","article-title":"A story of birth and death: mRNA translation and clearance at the onset of maternal-to-zygotic transition in mammals","volume":"101","author":"Sha","year":"2019","journal-title":"Biol. Reprod."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1007\/978-3-319-63187-5_3","article-title":"CHD1 Controls Cell Lineage Specification through Zygotic Genome Activation","volume":"229","author":"Suzuki","year":"2018","journal-title":"Adv. Anat. Embryol. Cell Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1262\/jrd.2015-029","article-title":"Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos","volume":"61","author":"Takahashi","year":"2015","journal-title":"J. Reprod. Dev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1744","DOI":"10.1101\/gad.1435106","article-title":"Maternal BRG1 regulates zygotic genome activation in the mouse","volume":"20","author":"Bultman","year":"2006","journal-title":"Genes Dev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1038\/ng.3858","article-title":"DUX-family transcription factors regulate zygotic genome activation in placental mammals","volume":"49","author":"Coluccio","year":"2017","journal-title":"Nat. Genet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1101\/gad.321174.118","article-title":"Dppa2 and Dppa4 directly regulate the Dux-driven zygotic transcriptional program","volume":"33","author":"Blotenburg","year":"2019","journal-title":"Genes Dev."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Yan, Y.L., Zhang, C., Hao, J., Wang, X.L., Ming, J., Mi, L., Na, J., Hu, X., and Wang, Y. (2019). DPPA2\/4 and SUMO E3 ligase PIAS4 opposingly regulate zygotic transcriptional program. PLoS Biol., 17.","DOI":"10.1371\/journal.pbio.3000324"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.anireprosci.2018.09.012","article-title":"Apoptosis and developmental capacity of vitrified parthenogenetic pig blastocysts","volume":"198","author":"Chen","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1072","DOI":"10.1071\/RD13405","article-title":"Effects of vitrification on the expression of pluripotency, apoptotic and stress genes in in vitro-produced porcine blastocysts","volume":"27","author":"Yeste","year":"2015","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1292","DOI":"10.1016\/j.theriogenology.2007.08.030","article-title":"Gene expression and development of mouse zygotes following droplet vitrification","volume":"68","author":"Dhali","year":"2007","journal-title":"Theriogenology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1017\/S0967199406003649","article-title":"Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts","volume":"14","author":"Park","year":"2006","journal-title":"Zygote"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1095\/biolreprod66.1.112","article-title":"Birth of piglets derived from porcine zygotes cultured in a chemically defined medium","volume":"66","author":"Yoshioka","year":"2002","journal-title":"Biol. Reprod."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1016\/0012-1606(89)90061-4","article-title":"Regulation of mouse preimplantation development: Inhibition of synthesis of proteins in the two-cell embryo that require transcription by inhibitors of cAMP-dependent protein kinase","volume":"133","author":"Poueymirou","year":"1989","journal-title":"Dev. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1298","DOI":"10.1111\/asj.12779","article-title":"Effects of melatonin on maturation, histone acetylation, autophagy of porcine oocytes and subsequent embryonic development","volume":"88","author":"Chen","year":"2017","journal-title":"Anim. Sci. J."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Cao, Z.B., Li, Y.S., Chen, Z., Wang, H., Zhang, M.L., Zhou, N.R., Wu, R., Ling, Y., Fang, F., and Li, N. (2015). Genome-Wide Dynamic Profiling of Histone Methylation during Nuclear Transfer-Mediated Porcine Somatic Cell Reprogramming. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0144897"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1262\/jrd.2019-094","article-title":"Vitrification of porcine cumulus-oocyte complexes at the germinal vesicle stage does not trigger apoptosis in oocytes and early embryos, but activates anti-apoptotic Bcl-XL gene expression beyond the 4-cell stage","volume":"66","author":"Somfai","year":"2020","journal-title":"J. Reprod. Develop."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"4883","DOI":"10.1242\/dev.01336","article-title":"The CCCH tandem zinc-finger protein Zfp36l2 is crucial for female fertility and early embryonic development","volume":"131","author":"Ramos","year":"2004","journal-title":"Development"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1007\/s10815-014-0172-9","article-title":"Melatonin modulates the expression of BCL-xl and improve the development of vitrified embryos obtained by IVF in mice","volume":"31","author":"Salehi","year":"2014","journal-title":"J. Assist. Reprod. Genet."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1007\/s11684-020-0792-7","article-title":"Effects of vitrification and cryostorage duration on single-cell RNA-Seq profiling of vitrified-thawed human metaphase II oocytes","volume":"15","author":"Huo","year":"2020","journal-title":"Front. Med."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/S0925-4773(99)00082-9","article-title":"Expression of Fgfr2 in the early mouse embryo indicates its involvement in preimplantation development","volume":"85","author":"Gorivodsky","year":"1999","journal-title":"Mech. Dev."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1016\/j.bbrc.2017.05.087","article-title":"Sirtuin inhibition leads to autophagy and apoptosis in porcine preimplantation blastocysts","volume":"488","author":"Kim","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.cryobiol.2019.01.012","article-title":"Vitrification of murine mature metaphase II oocytes perturbs DNA methylation reprogramming during preimplantation embryo development","volume":"87","author":"Cao","year":"2019","journal-title":"Cryobiology"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1007\/s10616-014-9838-5","article-title":"Histone methyltransferase SMYD3 regulates the expression of transcriptional factors during bovine oocyte maturation and early embryonic development","volume":"68","author":"Bai","year":"2016","journal-title":"Cytotechnology"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Zhang, J., Zhang, S., Wang, Y., Cheng, H., Hao, L., Zhai, Y., Zhang, Z., An, X., Ma, X., and Zhang, X. (2017). Effect of TET inhibitor on bovine parthenogenetic embryo development. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0189542"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1093\/biolre\/iox037","article-title":"Role of chemokine (C-C motif) ligand 24 in spatial arrangement of the inner cell mass of the bovine embryo","volume":"96","author":"Perez","year":"2017","journal-title":"Biol. Reprod."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"4232","DOI":"10.1242\/dev.081901","article-title":"Gata2 provides an early anterior bias and uncovers a global positioning system for polarity in the amniote embryo","volume":"139","author":"Bertocchini","year":"2012","journal-title":"Development"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.anireprosci.2012.08.020","article-title":"Pig oocyte vitrification by Cryotop method and the activation of the apoptotic cascade","volume":"135","author":"Vallorani","year":"2012","journal-title":"Anim. Reprod. Sci."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/5\/610\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:29:42Z","timestamp":1760135382000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/5\/610"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,28]]},"references-count":53,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ani12050610"],"URL":"https:\/\/doi.org\/10.3390\/ani12050610","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,28]]}}}