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Xiangpeng Dai State Key Laboratory of Reproductive Biology, Graduate School of the Chinese Academy of Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China
State Key Laboratory of Reproductive Biology, Graduate School of the Chinese Academy of Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Jie Hao State Key Laboratory of Reproductive Biology, Graduate School of the Chinese Academy of Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China
State Key Laboratory of Reproductive Biology, Graduate School of the Chinese Academy of Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Qi Zhou State Key Laboratory of Reproductive Biology, Graduate School of the Chinese Academy of Sciences, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, People's Republic of China

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Many strategies have been established to improve the efficiency of somatic cell nuclear transfer (SCNT), but relatively few focused on improving culture conditions. The effect of different culture media on preimplantation development of mouse nuclear transfer embryos was investigated. A modified sequential media method, named D media (M16/KSOM and CZB-EG/KSOM), was successfully established that significantly improves SCNT embryo development. Our result demonstrated that while lacking any adverse effect on in vivo fertilized embryos, the D media dramatically improves the blastocyst development of SCNT embryos compared with other commonly used media, including KSOM, M16, CZB, and αMEM. Specifically, the rate of blastocyst formation was 62.3% for D1 (M16/KSOM) versus 10–30% for the other media. An analysis of media components indicated that removing EDTA and glutamine from the media can be beneficial for early SCNT embryo development. Our results suggest that in vitro culture environment plays an important role in somatic cell reprogramming, and D media represent the most efficient culture method reported to date to support mouse SCNT early embryo development in vitro.

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Cheng Peng Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, First Hospital, Jilin University, Changchun, China

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Zhuo Lv New Hope Fertility Center, New York, USA

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Tang Hai State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Chaoyang, Beijing, China

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Xiangpeng Dai Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, First Hospital, Jilin University, Changchun, China

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Qi Zhou Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education, First Hospital, Jilin University, Changchun, China
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Chaoyang, Beijing, China

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Trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, can significantly improve the reprogramming efficiency of somatic cells. However, whether TSA has a detrimental effect on other kinds of embryos is largely unknown because of the lack of integrated analysis of the TSA effect on natural fertilized embryos. To investigate the effect of TSA on mouse embryo development, we analyzed preimplantation and post-implantation development of in vivo, in vitro fertilized, and parthenogenetic embryos treated with TSA at different concentrations and durations. In vivo fertilized embryos appeared to be the most sensitive to TSA treatment among the three groups, and the blastocyst formation rate decreased sharply as TSA concentration and treatment time increased. TSA treatment also reduced the livebirth rate for in vivo fertilized embryos from 56.59 to 38.33% but did not significantly affect postnatal biological functions such as the pups’ reproductive performance and their ability for spatial learning and memory. Further analysis indicated that the acetylation level of H3K9 and H4K5 was enhanced by TSA treatment at low concentrations, while DNA methylation appeared to be also disturbed by TSA treatment only at high concentration. Thus, our data indicates that TSA has different effects on preimplantation embryonic development depending on the nature of the embryo’s reproductive origin, the TSA concentration and treatment time, whereas the effect of TSA at the indicated concentration on postnatal function was minor.

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Liang Wu State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China
State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Honghui Zhou State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Haiyan Lin State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Jianguo Qi State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Cheng Zhu State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Zhiying Gao State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Hongmei Wang State Key Laboratory of Reproductive Biology, Graduate School, Department of Obstetrics and Gynecology, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing 100101, People's Republic of China

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Until recently, the molecular pathogenesis of preeclampsia (PE) remained largely unknown. Reports have shown that circulating microRNAs (miRNAs) are promising novel biomarkers for cancer, pregnancy, tissue injury, and other conditions. The objective of this study was to identify differentially expressed miRNAs in plasma from severe preeclamptic pregnancies compared with plasma from normal pregnancies. By mature miRNA microarray analysis, 15 miRNAs, including 13 up- and two downregulated miRNAs, were screened to be differentially expressed in plasma from women with severe PE (sPE). Seven miRNAs, namely miR-24, miR-26a, miR-103, miR-130b, miR-181a, miR-342-3p, and miR-574-5p, were validated to be elevated in plasma from severe preeclamptic pregnancies by real-time quantitative stem-loop RT-PCR analysis. Gene ontology and pathway enrichment analyses revealed that these miRNAs were involved in specific biological process categories (including regulation of metabolic processes, regulation of transcription, and cell cycle) and signaling pathways (including the MAP kinase signaling pathway, the transforming growth factor-β signaling pathway, and pathways in cancer metastasis). This study presents, for the first time, the differential expression profile of circulating miRNAs in sPE patients. The seven elevated circulating miRNAs may play critical roles in the pathogenesis of sPE, and one or more of them may become potential markers for diagnosing sPE.

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Cai-Xia Yang INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Zichuan Liu INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Renaud Fleurot INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Pierre Adenot INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Véronique Duranthon INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Xavier Vignon INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Qi Zhou INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Jean-Paul Renard INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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Nathalie Beaujean INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France
INRA, ENVA, State Key Laboratory of Reproductive Biology, UMR 1198 Biologie du Developpement et Reproduction, F-78350 Jouy en Josas, France

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To investigate the embryonic genome organization upon fertilization and somatic cell nuclear transfer (SCNT), we tracked HP1β and CENP, two well-characterized protein markers of pericentric and centromeric compartments respectively, in four types of embryos produced by rabbit in vivo fertilization, rabbit parthenogenesis, rabbit-to-rabbit, and bovine-to-rabbit SCNT. In the interphase nuclei of rabbit cultured fibroblasts, centromeres and associated pericentric heterochromatin are usually isolated. Clustering into higher-order chromatin structures, such as the chromocenters seen in mouse and bovine somatic cells, could not be observed in rabbit fibroblasts. After fertilization, centromeres and associated pericentric heterochromatin are quite dispersed in rabbit embryos. The somatic-like organization is progressively established and completed only by the 8/16-cell stage, a stage that corresponds to major embryonic genome activation in this species. In SCNT embryos, pericentric heterochromatin distribution typical for rabbit and bovine somatic cells was incompletely reverted into the 1-cell embryonic form with remnants of heterochromatin clusters in 100% of bovine-to-rabbit embryos. Subsequently, the donor cell nuclear organization was rapidly re-established by the 4-cell stage. Remarkably, the incomplete remodeling of bovine-to-rabbit 1-cell embryos was associated with delayed transcriptional activation compared with rabbit-to-rabbit embryos. Together, the results confirm that pericentric heterochromatin spatio-temporal reorganization is an important step of embryonic genome reprogramming. It also appears that genome reorganization in SCNT embryos is mainly dependent on the nuclear characteristics of the donor cells, not on the recipient cytoplasm.

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Yanfang Wu College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Zhenzi Zuo State Key Laboratory of Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing, P.R. China

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Zheng Wang College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Hanghang Liu College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Qi Zhou College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Subi Ren College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Xinrui Lan College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Yong Zhang College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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Yongsheng Wang College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, P.R. China
Key Laboratory of Animal Biotechnology, Ministry of Agriculture and Rural Affairs, Northwest A&F University, Yangling, P.R. China

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In brief

Almost total lack of sperm-borne RNAs is regarded as one of the key factors that leads to the abnormal development of somatic cell nuclear transfer embryo. This paper reveals a need for us to further explore the roles of the paternal regulatory factors on embryonic development in early embryos.

Abstract

Mature sperm contain both coding and non-coding RNAs, which can be delivered into an oocyte with the sperm at fertilization. Accumulating evidences show that these sperm-borne RNAs play crucial roles in epigenetic reprogramming, cytoskeleton remodeling, embryonic development, and offspring phenotype. Almost total lack of sperm-borne RNAs is regarded as one of the key factors that leads to the abnormal development of somatic cell nuclear transfer (SCNT) embryo. bta-miR-183 was found to be highly expressed in bovine sperm and can be delivered into oocytes during fertilization in our previous study, and in this study, EZR was confirmed as a target gene of bta-miR-183 in early embryos by bioinformatics, luciferase, and gain-of-function and loss-of-function experiments. Scanning electron microscopy showed that the density of microvilli on the surface of SCNT embryos was significantly higher than that onin vitro fertilized embryos and was significantly reduced by injection of bta-miR-183 mimic. EZR-siRNA injected into SCNT embryos had a similar effect. This indicated that the lack of bta-miR-183 might lead to abnormal changes in microvilli by downregulating ezrin protein. In addition, gain-of-function studies showed that bta-miR-183 significantly improved developmental competence of SCNT embryo in terms of cleavage (76.63% vs 64.32%, P < 0.05), blastocyst formation (43.75% vs 28.26%, P < 0.05), apoptotic index (5.21% vs 12.64%, P < 0.05), and the trophoblast ratio (32.65% vs 25.58%, P < 0.05) in day 7 blastocysts. Thus, the present study indicated that bta-miR-183 might play crucial roles in the formation of microvilli and embryo development by regulating expression of EZR mRNA.

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Yang Yu State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Chenhui Ding State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Eryao Wang State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Xinjie Chen State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Xuemei Li State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Chunli Zhao State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Yong Fan State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Liu Wang State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Nathalie Beaujean State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Qi Zhou State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Alice Jouneau State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Weizhi Ji State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China, Department of Reproduction and Development, Kunming Institute of Zoology and Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan, China, Graduate University of Chinese Academy of Sciences, Beijing, China and INRA, UMR 1198; ENVA; CNRS, FRE 2857, Biologie du Développement et Reproduction, Jouy en Josas 78350, France

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Even though it generates healthy adults, nuclear transfer in mammals remains an inefficient process. Mainly attributed to abnormal reprograming of the donor chromatin, this inefficiency may also be caused at least partly by a specific effect of the cloning technique which has not yet been well investigated. There are two main procedures for transferring nuclei into enucleated oocytes: fusion and piezoelectric microinjection, the latter being used mostly in mice. We have, therefore, decided to compare the quality and the developmental ability, both in vivo and in vitro, of embryos reconstructed with electrofusion or piezoelectric injection. In addition, the effect of piezo setups of differing electric strengths was investigated. Along with the record of the rate of development, we compared the nuclear integrity in the blastomeres during the first cleavages as well as the morphological and cellular quality of the blastocysts. Our results show that the piezo-assisted micromanipulation can induce DNA damage in the reconstructed embryos, apoptosis, and reduced cell numbers in blastocysts as well as a lower rate of development to term. Even if piezo-driven injection facilitates a faster and more efficient rate of reconstruction, it should be used with precaution and with as low parameters as possible.

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Da Li Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, China

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Yue You Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, China

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Fang-Fang Bi Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, China

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Tie-Ning Zhang Department of Pediatrics, Shengjing Hospital of China Medical University, Shenyang, China

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Jiao Jiao Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, China

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Tian-Ren Wang Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, China
Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale School of Medicine, New Haven, Connecticut, USA

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Yi-Ming Zhou Department of Medicine, Brigham and Women’s Hospital, Harvard Institutes of Medicine, Harvard Medical School, Boston, Massachusetts, USA

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Zi-Qi Shen Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, China

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Xiu-Xia Wang Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, China

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Qing Yang Department of Obstetrics and Gynecology, Shengjing Hospital of China Medical University, Shenyang, China

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The importance of autophagy in polycystic ovary syndrome (PCOS)-related metabolic disorders is increasingly being recognized, but few studies have investigated the role of autophagy in PCOS. Here, transmission electron microscopy demonstrated that autophagy was enhanced in the ovarian tissue from both humans and rats with PCOS. Consistent with this, ovarian granulosa cells from PCOS rats showed increases in the autophagy marker protein light chain 3B (LC3B), whereas levels of the autophagy substrate SQSTM1/p62 were decreased. In addition, the ratio of LC3-II/LC3-I was markedly elevated in human PCOS ovarian tissue compared with normal ovarian tissue. Real-time PCR arrays indicated that 7 and 34 autophagy-related genes were down- and up-regulated in human PCOS , Signal-Net, and regression analysis suggested that there are a wide range of interactions among these 41 genes, and a potential network based on EGFR, ERBB2, FOXO1, MAPK1, NFKB1, IGF1, TP53 and MAPK9 may be responsible for autophagy activation in PCOS. Systematic functional analysis of 41 differential autophagy-related genes indicated that these genes are highly involved in specific cellular processes such as response to stress and stimulus, and are linked to four significant pathways, including the insulin, ERBB, mTOR signaling pathways and protein processing in the endoplasmic reticulum. This study provides evidence for a potential role of autophagy disorders in PCOS in which autophagy may be an important molecular event in the pathogenesis of PCOS.

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