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Zona pellucida removal modifies the expression and release of specific microRNAs in domestic cat blastocysts

Published online by Cambridge University Press:  19 September 2023

Daniel Veraguas-Dávila Open the ORCID record for Daniel Veraguas-Dávila [Opens in a new window] ,
Diego Caamaño ,
Darling Saéz-Ruiz ,
Yazmín Vásquez ,
Fernando Saravia ,
Fidel Ovidio Castro  and
Lleretny Rodríguez-Alvarez
Daniel Veraguas-Dávila*
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile Facultad de Ciencias Agrarias y Forestales, Departamento de Ciencias Agrarias, Escuela de Medicina Veterinaria, Universidad Católica del Maule, Los Niches, Curicó, Chile
Diego Caamaño
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
Darling Saéz-Ruiz
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
Yazmín Vásquez
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
Fernando Saravia
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
Fidel Ovidio Castro
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
Lleretny Rodríguez-Alvarez
Affiliation:
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán, Chile
*
Corresponding author: Daniel Veraguas-Dávila; Email: dveraguas@ucm.cl
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Summary

The in vitro culture of domestic cat embryos without the zona pellucida affects their implantation capacity. MicroRNAs (miRNAs) have an important role in embryo–maternal communication and implantation. The objective of this study was to evaluate the expression of specific miRNAs in domestic cat blastocysts cultured without the zona pellucida. Two experimental groups were done: (1) domestic cat embryos cultured with the zona pellucida (zona intact control group, ZI); and (2) cultured without the zona pellucida (zona free group, ZF). The cleavage, morula and blastocyst rates were evaluated. The blastocysts and their spent medium were used for miRNA expression analysis using RT-qPCR (miR-21, miR-24, mi25, miR-29, miR-96, miR-98, miR-103, miR-191, miR-196, miR-199, miR-130, miR-155 and miR-302). The pre-mature microRNAs (pre-miRNAs) and miRNAs were evaluated in the blastocysts and only miRNAs were evaluated in the spent medium. No differences were observed in the cleavage, morula and blastocyst rates between the ZF and ZI groups (P > 0.05). For miRNAs analysis, miR-103 and miR-191 had the most stable expression and were selected as internal controls. ZF blastocysts had a higher expression of miR-21, miR-25, miR-29 and miR-199 and a lower expression of miR-96 than their ZI counterparts (P < 0.05). Furthermore, higher levels of miR-21, miR-25 and miR-98 were detected in the spent medium of ZF blastocysts (P < 0.05). In conclusion, in vitro culture of domestic cat embryos without the zona pellucida modifies the expression of miR-21, miR-25, miR-29, miR-199 and miR-96 at the blastocyst stage and the release of miR-21, miR-25 and miR-98.

Keywords

Felid embryosGene expressionIn vitro fertilizationMicroRNAs
Type
Research Article
Information
Zygote , Volume 31 , Issue 6 , December 2023 , pp. 544 - 556
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Copyright
© The Author(s), 2023. Published by Cambridge University Press

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References

Abu-Halima, M., Häusler, S., Backes, C., Fehlmann, T., Staib, C., Nestel, S., Nazarenko, I., Meese, E. and Keller, A. (2017). Micro-ribonucleic acids and extracellular vesicles repertoire in the spent culture media is altered in women undergoing in vitro fertilization. Scientific Reports, 7(1), 13525. doi: 10.1038/s41598-017-13683-8 CrossRefGoogle ScholarPubMed
Andersen, C. L., Jensen, J. L. and Ørntoft, T. F. (2004). Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Research, 64(15), 5245–5250. doi: 10.1158/0008-5472.CAN-04-0496 CrossRefGoogle Scholar
Andrade, G. M., Bomfim, M. M., Del Collado, M., Meirelles, F. V., Perecin, F. and da Silveira, J. C. (2019). Oxygen tension modulates extracellular vesicles and its miRNA contents in bovine embryo culture medium. Molecular Reproduction and Development, 86(8), 1067–1080. doi: 10.1002/mrd.23223 CrossRefGoogle ScholarPubMed
Balcells, I., Cirera, S. and Busk, P. K. (2011). Specific and sensitive quantitative RT-PCR of miRNAs with DNA primers. BMC Biotechnology, 11, 70. doi: 10.1186/1472-6750-11-70 CrossRefGoogle ScholarPubMed
Buemo, C. P., Gambini, A., Moro, L. N., Hiriart, M. I., Fernández-Martín, R., Collas, P. and Salamone, D. F. (2016). Embryo aggregation in pig improves cloning efficiency and embryo quality. PLOS ONE, 11(2), e0146390. doi: 10.1371/journal.pone.0146390 CrossRefGoogle ScholarPubMed
Busk, P. K. (2014). A tool for design of primers for microRNA-specific quantitative RT-qPCR. BMC Bioinformatics, 15, 29. doi: 10.1186/1471-2105-15-29 CrossRefGoogle ScholarPubMed
Capalbo, A., Ubaldi, F. M., Cimadomo, D., Noli, L., Khalaf, Y., Farcomeni, A., Ilic, D. and Rienzi, L. (2016). MicroRNAs in spent blastocyst culture medium are derived from trophectoderm cells and can be explored for human embryo reproductive competence assessment. Fertility and Sterility, 105(1), 225–35.e1. doi: 10.1016/j.fertnstert.2015.09.014 CrossRefGoogle ScholarPubMed
Dehghan, Z., Mohammadi-Yeganeh, S., Rezaee, D. and Salehi, M. (2021). MicroRNA-21 is involved in oocyte maturation, blastocyst formation, and pre-implantation embryo development. Developmental Biology, 480, 69–77. doi: 10.1016/j.ydbio.2021.08.008 CrossRefGoogle ScholarPubMed
Denker, H. W. (2000). Structural dynamics and function of early embryonic coats. Cells, Tissues, Organs, 166(2), 180–207. doi: 10.1159/000016732 CrossRefGoogle ScholarPubMed
Doridot, L., Miralles, F., Barbaux, S. and Vaiman, D. (2013). Trophoblasts, invasion, and microRNA. Frontiers in Genetics, 4, 248. doi: 10.3389/fgene.2013.00248 CrossRefGoogle ScholarPubMed
Fan, W., Huang, T., Wu, T., Bai, H., Kawahara, M. and Takahashi, M. (2022). Zona pellucida removal by acid Tyrode’s solution affects pre-and post-implantation development and gene expression in mouse embryos. Biology of Reproduction, 107(5), 1228–1241. doi: 10.1093/biolre/ioac155 Google ScholarPubMed
Forero, D. A., González-Giraldo, Y., Castro-Vega, L. J. and Barreto, G. E. (2019). qPCR-based methods for expression analysis of miRNAs. BioTechniques, 67(4), 192–199. doi: 10.2144/btn-2019-0065 CrossRefGoogle ScholarPubMed
Gambini, A., Jarazo, J., Olivera, R. and Salamone, D. F. (2012). Equine cloning: In vitro and in vivo development of aggregated embryos. Biology of Reproduction, 87(1), 15–11. doi: 10.1095/biolreprod.112.098855 CrossRefGoogle ScholarPubMed
Gómez, M. C., Pope, C. E., Ricks, D. M., Lyons, J., Dumas, C. and Dresser, B. L. (2009). Cloning endangered felids using heterospecific donor oocytes and interspecies embryo transfer. Reproduction, Fertility, and Development, 21(1), 76–82. doi: 10.1071/rd08222 CrossRefGoogle ScholarPubMed
Gong, M., Yu, B., Wang, J., Wang, Y., Liu, M., Paul, C., Millard, R. W., Xiao, D. S., Ashraf, M. and Xu, M. (2017). Mesenchymal stem cells release exosomes that transfer miRNAs to endothelial cells and promote angiogenesis. Oncotarget, 8(28), 45200–45212. doi: 10.18632/oncotarget.16778 CrossRefGoogle ScholarPubMed
Gross, N., Kropp, J. and Khatib, H. (2017a). MicroRNA signaling in embryo development. Biology, 6(3), 34. doi: 10.3390/biology6030034 CrossRefGoogle ScholarPubMed
Gross, N., Kropp, J. and Khatib, H. (2017b). Sexual dimorphism of miRNAs secreted by bovine in vitro-produced embryos. Frontiers in Genetics, 8, 39. doi: 10.3389/fgene.2017.00039 CrossRefGoogle ScholarPubMed
Hawke, D. C., Ahmed, D. B., Watson, A. J. and Betts, D. H. (2021a). Murine blastocysts release mature microRNAs into culture media that reflect developmental status. Frontiers in Genetics, 12, 655882. doi: 10.3389/fgene.2021.655882 CrossRefGoogle ScholarPubMed
Hawke, D. C., Watson, A. J. and Betts, D. H. (2021b). Extracellular vesicles, microRNA and the preimplantation embryo: Non-invasive clues of embryo well-being. Reproductive Biomedicine Online, 42(1), 39–54. doi: 10.1016/j.rbmo.2020.11.011 CrossRefGoogle ScholarPubMed
Herrler, A. and Beier, H. M. (2000). Early embryonic coats: Morphology, function, practical applications. An overview. Cells, Tissues, Organs, 166(2), 233–246. doi: 10.1159/000016736 CrossRefGoogle ScholarPubMed
Hysolli, E., Tanaka, Y., Su, J., Kim, K. Y., Zhong, T., Janknecht, R., Zhou, X. L., Geng, L., Qiu, C., Pan, X., Jung, Y. W., Cheng, J., Lu, J., Zhong, M., Weissman, S. M. and Park, I. H. (2016). Regulation of the DNA methylation landscape in human somatic cell reprogramming by the miR-29 family. Stem Cell Reports, 7(1), 43–54. doi: 10.1016/j.stemcr.2016.05.014 CrossRefGoogle ScholarPubMed
Kanda, M., Oikawa, H., Nakao, H. and Tsutsui, T. (1995). Early embryonic development in vitro and embryo transfer in the cat. Journal of Veterinary Medical Science, 57(4), 641–646. doi: 10.1292/jvms.57.641 CrossRefGoogle ScholarPubMed
Kim, J., Lee, J., Lee, T. B. and Jun, J. H. (2019). Embryotrophic effects of extracellular vesicles derived from outgrowth embryos in pre- and peri-implantation embryonic development in mice. Molecular Reproduction and Development, 86(2), 187–196. doi: 10.1002/mrd.23093 CrossRefGoogle ScholarPubMed
Krol, J., Loedige, I. and Filipowicz, W. (2010). The widespread regulation of microRNA biogenesis, function and decay. Nature Reviews. Genetics, 11(9), 597–610. doi: 10.1038/nrg2843 CrossRefGoogle ScholarPubMed
Kropp, J. and Khatib, H. (2015). Characterization of microRNA in bovine in vitro culture media associated with embryo quality and development. Journal of Dairy Science, 98(9), 6552–6563. doi: 10.3168/jds.2015-9510 CrossRefGoogle ScholarPubMed
Kropp, J., Salih, S. M. and Khatib, H. (2014). Expression of microRNAs in bovine and human pre-implantation embryo culture media. Frontiers in Genetics, 5, 91. doi: 10.3389/fgene.2014.00091 CrossRefGoogle ScholarPubMed
Liang, J., Wang, S. and Wang, Z. (2017). Role of microRNAs in embryo implantation. Reproductive Biology and Endocrinology: RB&E, 15(1), 90. doi: 10.1186/s12958-017-0309-7 CrossRefGoogle ScholarPubMed
Liang, S., Nie, Z. W., Guo, J., Niu, Y. J., Shin, K. T., Ock, S. A. and Cui, X. S. (2018). Overexpression of microRNA-29b decreases expression of DNA methyltransferases and improves quality of the blastocysts derived from somatic cell nuclear transfer in cattle. Microscopy and Microanalysis, 24(1), 29–37. doi: 10.1017/S1431927618000016 CrossRefGoogle ScholarPubMed
Liu, W.-M., Pang, R. T.K., Cheong, A.W.Y., Ng, E. H. Y., Lao, K., Lee, K-F. and Yeung, W. S. B. (2012). Involvement of microRNA lethal-7a in the regulation of embryo implantation in mice. PLoS One, 7(5), e37039. doi: 10.1371/journal.pone.0037039.CrossRefGoogle ScholarPubMed
Liu, W., Niu, Z., Li, Q., Pang, R. T., Chiu, P. C. and Yeung, W. S.-B. (2016). MicroRNA and embryo implantation. American Journal of Reproductive Immunology, 75(3), 263–271. doi: 10.1111/aji.12470 CrossRefGoogle ScholarPubMed
Lunn, M. O. and Wright, S. J. (2009). Imaging the zona pellucida of canine and feline oocytes using scanning electron microscopy. Microscopy and Microanalysis, 15(1), 2–14. doi: 10.1017/S1431927609090084 CrossRefGoogle ScholarPubMed
Lv, C., Yu, W. X., Wang, Y., Yi, D. J., Zeng, M. H. and Xiao, H. M. (2018). MiR-21 in extracellular vesicles contributes to the growth of fertilized eggs and embryo development in mice. Bioscience Reports, 38(4). doi: 10.1042/BSR20180036 CrossRefGoogle Scholar
Mahdipour, M., van Tol, H. T., Stout, T. A. and Roelen, B. A. (2015). Validating reference microRNAs for normalizing qRT-PCR data in bovine oocytes and preimplantation embryos. BMC Developmental Biology, 15, 25. doi: 10.1186/s12861-015-0075-8 CrossRefGoogle ScholarPubMed
Mohammed, B. T., Sontakke, S. D., Ioannidis, J., Duncan, W. C. and Donadeu, F. X. (2017). The adequate corpus luteum: miR-96 promotes luteal cell survival and progesterone production. Journal of Clinical Endocrinology and Metabolism, 102(7), 2188–2198. doi: 10.1210/jc.2017-00259 CrossRefGoogle ScholarPubMed
Moro, L. N., Jarazo, J., Buemo, C., Hiriart, M. I., Sestelo, A. and Salamone, D. F. (2015a). Tiger, Bengal and domestic cat embryos produced by homospecific and interspecific zona-free nuclear transfer. Reproduction in Domestic Animals, 50(5), 849–857. doi: 10.1111/rda.12593 CrossRefGoogle ScholarPubMed
Moro, L. N., Hiriart, M. I., Buemo, C., Jarazo, J., Sestelo, A., Veraguas, D., Rodriguez-Alvarez, L. and Salamone, D. F. (2015b). Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression. Reproduction, 150(1), 1–10. doi: 10.1530/REP-15-0048 CrossRefGoogle Scholar
Nakamura, K., Kusama, K., Ideta, A., Kimura, K., Hori, M. and Imakawa, K. (2019). Effects of miR-98 in intrauterine extracellular vesicles on maternal immune regulation during the peri-implantation period in cattle. Scientific Reports, 9(1), 20330. doi: 10.1038/s41598-019-56879-w CrossRefGoogle ScholarPubMed
Paul, A. B. M., Sadek, S. T. and Mahesan, A. M. (2019). The role of microRNAs in human embryo implantation: A review. Journal of Assisted Reproduction and Genetics, 36(2), 179–187. doi: 10.1007/s10815-018-1326-y CrossRefGoogle ScholarPubMed
Pohler, K. G., Green, J. A., Moley, L. A., Gunewardena, S., Hung, W. T., Payton, R. R., Hong, X., Christenson, L. K., Geary, T. W. and Smith, M. F. (2017) Circulating microRNA as candidates for early embryonic viability in cattle. Molecular Reproduction and Development, 84(8), 731–743. doi: 10.1002/mrd.22856 CrossRefGoogle ScholarPubMed
Pope, C. E. (2014). Aspects of in vivo oocyte production, blastocyst development, and embryo transfer in the cat. Theriogenology, 81(1), 126–137. doi: 10.1016/j.theriogenology.2013.09.006 CrossRefGoogle ScholarPubMed
Pope, C. E. (2019a). Forty years of assisted reproduction research in nondomestic, wild and endangered mammals. Revista Brasileira de Reprodução Animal, 43(2), 160–167.Google Scholar
Pope, C. E. (2019b). Thirty years of assisted reproductive technology in the domestic cat: A selected summary. Revista Brasileira de Reprodução Animal, 43(2), 129–136.Google Scholar
Prasad, S. V., Skinner, S. M., Carino, C., Wang, N., Cartwright, J. and Dunbar, B. S. (2000). Structure and function of the proteins of the mammalian zona pellucida. Cells, Tissues, Organs, 166(2), 148–164. doi: 10.1159/000016730 CrossRefGoogle ScholarPubMed
Reza, A. M. M. T., Choi, Y. J., Han, S. G., Song, H., Park, C., Hong, K. and Kim, J. H. (2019). Roles of microRNAs in mammalian reproduction: From the commitment of germ cells to peri-implantation embryos. Biological Reviews of the Cambridge Philosophical Society, 94(2), 415–438. doi: 10.1111/brv.12459 CrossRefGoogle Scholar
Rodríguez-Alvarez, L., Sharbati, J., Sharbati, S., Cox, J. F., Einspanier, R. and Castro, F. O. (2010). Differential gene expression in bovine elongated (Day 17) embryos produced by somatic cell nucleus transfer and in vitro fertilization. Theriogenology, 74(1), 45–59. doi: 10.1016/j.theriogenology.2009.12.018 CrossRefGoogle ScholarPubMed
Shen, X. H., Han, Y. J., Zhang, D. X., Cui, X. S. and Kim, N. H. (2009). A link between the interleukin-6/Stat3 anti-apoptotic pathway and microRNA-21 in preimplantation mouse embryos. Molecular Reproduction and Development, 76(9), 854–862. doi: 10.1002/mrd.21048 CrossRefGoogle ScholarPubMed
Tan, K., Wang, X., Zhang, Z., Miao, K., Yu, Y., An, L. and Tian, J. (2016). Downregulation of miR-199a-5p disrupts the developmental potential of in vitro-fertilized mouse blastocysts. Biology of Reproduction, 95(3), 54. doi: 10.1095/biolreprod.116.141051 CrossRefGoogle ScholarPubMed
Trohatou, O., Zagoura, D., Bitsika, V., Pappa, K. I., Antsaklis, A., Anagnou, N. P. and Roubelakis, M. G. (2014). Sox2 suppression by miR-21 governs human mesenchymal stem cell properties. Stem Cells Translational Medicine, 3(1), 54–68. doi: 10.5966/sctm.2013-0081 CrossRefGoogle ScholarPubMed
Turchinovich, A., Samatov, T. R., Tonevitsky, A. G. and Burwinkel, B. (2013). Circulating miRNAs: Cell–cell communication function? Frontiers in Genetics, 4, 119. doi: 10.3389/fgene.2013.00119 CrossRefGoogle ScholarPubMed
Vajta, G., Korösi, T., Du, Y., Nakata, K., Ieda, S., Kuwayama, M. and Nagy, Z. P. (2008). The Well-of-the-well system: An efficient approach to improve embryo development. Reproductive Biomedicine Online, 17(1), 73–81. doi: 10.1016/s1472-6483(10)60296-9 CrossRefGoogle ScholarPubMed
Veraguas, D., Echeverry, D., Castro, F. O. and Rodriguez-Alvarez, L. (2017a) Applied biotechnologies in the conservation of wild felids: In vitro embryo production and cellular regenerative therapies. In A. B. Shrivastav & K. P. Singh (Eds.) Big cats IntechOpen; Chapter 4. doi: 10.5772/intechopen.71311 CrossRefGoogle Scholar
Veraguas, D., Gallegos, P. F., Castro, F. O. and Rodriguez-Alvarez, L. (2017b). Cell cycle synchronization and analysis of apoptosis-related gene in skin fibroblasts from domestic cat (Felis silvestris catus) and kodkod (Leopardus guigna). Reproduction in Domestic Animals, 52(5), 881–889. doi: 10.1111/rda.12994 CrossRefGoogle ScholarPubMed
Veraguas, D., Gallegos, P. F., Velasquez, A. E., Castro, F. O. and Rodriguez-Alvarez, L. (2017c). FSH stimulation of anestrous cats improves oocyte quality and development of parthenogenetic embryos. Theriogenology, 87, 25–35. doi: 10.1016/j.theriogenology.2016.08.008 CrossRefGoogle ScholarPubMed
Veraguas, D., Cuevas, S. R., Gallegos, P. F., Saez-Ruiz, D., Castro, F. O. and Rodriguez-Alvarez, L. (2018). eCG stimulation in domestic cats increases the expression of gonadotrophin-induced genes improving oocyte competence during the non-breeding season. Reproduction in Domestic Animals, 53(6), 1306–1316. doi: 10.1111/rda.13229 CrossRefGoogle ScholarPubMed
Veraguas, D., Saez, S., Aguilera, C., Echeverry, D., Gallegos, P. F., Saez-Ruiz, D., Castro, F. O. and Rodriguez-Alvarez, L. (2020a). In vitro and in vivo development of domestic cat embryos generated by in vitro fertilization after eCG priming and oocyte in vitro maturation. Theriogenology, 146, 94–103. doi: 10.1016/j.theriogenology.2020.02.012 CrossRefGoogle ScholarPubMed
Veraguas, D., Aguilera, C., Echeverry, D., Saez-Ruiz, D., Castro, F. O. and Rodriguez-Alvarez, L. (2020b). Embryo aggregation allows the production of kodkod (Leopardus guigna) blastocysts after interspecific SCNT. Theriogenology, 158, 148–157. doi: 10.1016/j.theriogenology.2020.09.006 CrossRefGoogle ScholarPubMed
Veraguas-Davila, D., Cordero, M. F., Saez, S., Saez-Ruiz, D., Gonzalez, A., Saravia, F., Castro, F. O. and Rodriguez-Alvarez, L. (2021). Domestic cat embryos generated without zona pellucida are capable of developing in vitro but exhibit abnormal gene expression and a decreased implantation rate. Theriogenology, 174, 36–46. doi: 10.1016/j.theriogenology.2021.08.013 CrossRefGoogle Scholar
Veraguas-Dávila, D., Saéz-Ruíz, D., Álvarez, M. C., Saravia, F., Castro, F. O. and Rodríguez-Alvarez, L. (2022). Analysis of trophectoderm markers in domestic cat blastocysts cultured without zona pellucida. Zygote, 30(6), 841–848. doi: 10.1017/S096719942200034X CrossRefGoogle ScholarPubMed
Viswanathan, S. R., Mermel, C. H., Lu, J., Lu, C. W., Golub, T. R. and Daley, G. Q. (2009). microRNA expression during trophectoderm specification. PLOS ONE, 4(7), e6143. doi: 10.1371/journal.pone.0006143 CrossRefGoogle ScholarPubMed
Wang, Y., Zhou, T., Wan, J., Yang, Y., Chen, X., Wang, J., Zhou, C., Liu, M., Ling, X. and Zhang, J. (2016). Comparative transcriptome analysis reveals a regulatory network of microRNA-29b during mouse early embryonic development. Oncotarget, 7(33), 53772–53782. doi: 10.18632/oncotarget.10741 CrossRefGoogle ScholarPubMed
Xia, H. F., Jin, X. H., Cao, Z. F., Hu, Y. and Ma, X. (2014a). MicroRNA expression and regulation in the uterus during embryo implantation in rat. FEBS Journal, 281(7), 1872–1891. doi: 10.1111/febs.12751 CrossRefGoogle ScholarPubMed
Xia, H. F., Jin, X. H., Cao, Z. F., Shi, T. and Ma, X. (2014b). MiR-98 is involved in rat embryo implantation by targeting Bcl-xL. FEBS Letters, 588(4), 574–583. doi: 10.1016/j.febslet.2013.12.026 CrossRefGoogle ScholarPubMed
Yang, Y., Xie, Y., Wu, M., Geng, Y., Li, R., Xu, L., Liu, X. and Pan, Y. (2017). Expression of mmu-miR-96 in the endometrium during early pregnancy and its regulatory effects on stromal cell apoptosis via Bcl2. Molecular Medicine Reports, 15(4), 1547–1554. doi: 10.3892/mmr.2017.6212 CrossRefGoogle ScholarPubMed
Zhang, C., Shi, Y. R., Liu, X. R., Cao, Y. C., Tian, J. L., Jia, Z. Y., Zhen, D., Liu, F. H. and Gao, J. M. (2014). The regulatory role of icariin on apoptosis in mouse preimplantation embryos with reduced microRNA-21. Theriogenology, 82(3), 461–468. doi: 10.1016/j.theriogenology.2014.05.006 CrossRefGoogle ScholarPubMed
Zhang, Y., Feng, G. H., Xu, K., Wang, L., Cui, P., Li, Y., Wang, C., Teng, F., Hao, J., Wan, H. F., Tan, Y., Wang, X. J. and Zhou, Q. (2016). A non-invasive method to determine the pluripotent status of stem cells by culture medium microRNA expression detection. Scientific Reports, 6(1), 22380. doi: 10.1038/srep22380 CrossRefGoogle ScholarPubMed