PMID- 28643089 OWN - NLM STAT- MEDLINE DCOM- 20180709 LR - 20181113 IS - 1573-7330 (Electronic) IS - 1058-0468 (Print) IS - 1058-0468 (Linking) VI - 34 IP - 9 DP - 2017 Sep TI - Time-lapse observation and transcriptome analysis of a case with repeated multiple pronuclei after IVF/ICSI. PG - 1189-1197 LID - 10.1007/s10815-017-0972-9 [doi] AB - PURPOSE: The purpose of this study was to investigate the cause of repeated multipronucleus (MPN) formation in zygotes in a patient after both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHOD: This is a case study. A patient had unexplained primary infertility with recurring total MPN zygotes after IVF and ICSI cycles. Time-lapse monitoring of pronucleus formation was carried out. Embryos developed from MPN zygotes were analyzed by fluorescence in situ hybridization (FISH). Single-cell RNA-seq analysis was used to identify gene expression profiles of the patient's oocyte and zygote, and these were compared to the data from oocytes and zygotes from donors with normal fertilization (patient, n = 1; donors, n = 4). Oocyte-specific genes with differential expression were selected by the Amazonia! RESULTS: From time-lapse analysis, we observed the formation of multiple micronuclei near the site of the second polar body extrusion. These micronuclei migrated, expanded, and juxtaposed with the male pronucleus leading to a multipronucleus. None of these MPN zygotes could develop to the blastocyst stage, and FISH analysis revealed a chaotic chromosomal complement in the arrested embryos. RNA-seq analysis showed 113 differentially expressed genes (DEGs) between the patient and the donor oocytes and zygotes. Moreover, 25 of the 113 DEGs were unique or highly expressed in oocytes and early embryos. From 25 DEGs, three genes, DYNC2LI1, NEK2, and CCNH, which are involved in meiosis and the chromosome separation process, were further validated by real-time PCR. CONCLUSION: We identified several candidate genes affecting pronucleus formation as a new cause of infertility. FAU - Dai, J AU - Dai J AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. FAU - Leng, L Z AU - Leng LZ AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. FAU - Lu, C F AU - Lu CF AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. AD - National Engineering and Research Center of Human Stem Cell, Changsha, China. FAU - Gong, F AU - Gong F AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. AD - National Engineering and Research Center of Human Stem Cell, Changsha, China. FAU - Zhang, S P AU - Zhang SP AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. FAU - Zheng, W AU - Zheng W AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. FAU - Lu, G X AU - Lu GX AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. AD - National Engineering and Research Center of Human Stem Cell, Changsha, China. AD - Key Laboratory of Reproductive and Stem Cell Engineering, Ministry of Health, Changsha, China. FAU - Lin, G AU - Lin G AD - Reproductive and Genetic Hospital of CITIC-XIANGYA, Changsha, China. linggf@hotmail.com. AD - Institute of Reproductive and Stem Cell Engineering, Basic Medicine College, Central South University, Changsha, China. linggf@hotmail.com. AD - National Engineering and Research Center of Human Stem Cell, Changsha, China. linggf@hotmail.com. AD - Key Laboratory of Reproductive and Stem Cell Engineering, Ministry of Health, Changsha, China. linggf@hotmail.com. LA - eng GR - No. 2015zzts101/the Fundamental Research Funds for the Central Universities of Central South University/ GR - No. 81222007/the National Natural Science Foundation of China/ GR - No. 81471510/the National Natural Science Foundation of China/ GR - No. 907010003/Program for New Century Excellent Talents in University/ PT - Journal Article DEP - 20170622 PL - Netherlands TA - J Assist Reprod Genet JT - Journal of assisted reproduction and genetics JID - 9206495 SB - IM MH - Cell Nucleus/genetics/*ultrastructure MH - Female MH - *Fertilization in Vitro MH - *Gene Expression Profiling MH - Humans MH - In Situ Hybridization, Fluorescence MH - Infertility/genetics/pathology MH - Male MH - Meiosis/genetics MH - Oocytes/cytology MH - Sperm Injections, Intracytoplasmic/methods MH - Time-Lapse Imaging MH - Transcriptome/genetics MH - Zygote/metabolism/*ultrastructure PMC - PMC5581788 OTO - NOTNLM OT - Multipronuclear OT - Oocyte OT - Single-cell transcriptome OT - Time-lapse COIS- This study was approved by the Ethics Committee of the Reproductive and Genetic Hospital of CITIC-Xiangya (reference LL-SC-SG-2013-012). ETHICAL APPROVAL: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. INFORMED CONSENT: Informed consent was obtained from all individual participants included in the study. CONFLICT OF INTEREST: The authors declare that they have no conflict of interest. EDAT- 2017/06/24 06:00 MHDA- 2018/07/10 06:00 PMCR- 2018/09/01 CRDT- 2017/06/24 06:00 PHST- 2017/03/01 00:00 [received] PHST- 2017/06/07 00:00 [accepted] PHST- 2017/06/24 06:00 [pubmed] PHST- 2018/07/10 06:00 [medline] PHST- 2017/06/24 06:00 [entrez] PHST- 2018/09/01 00:00 [pmc-release] AID - 10.1007/s10815-017-0972-9 [pii] AID - 972 [pii] AID - 10.1007/s10815-017-0972-9 [doi] PST - ppublish SO - J Assist Reprod Genet. 2017 Sep;34(9):1189-1197. doi: 10.1007/s10815-017-0972-9. Epub 2017 Jun 22.