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pronuclear embryo

P Thuwanut, W Tipkantha, B Siriaroonrat, P Comizzoli, K Chatdarong
The Indochinese leopard (Panthera pardus delacouri) population, included in CITES Appendix I, has been declining for decades. Proper gamete preservation condition is critical for breeding programme management using artificial insemination or in vitro fertilization (IVF). The present study aimed at investigating the impact of post-thawing treatment of leopard semen with extracellular adenosine 5'-triphosphate (ATPe) on sperm quality (including morphological traits and ability to fertilize an oocyte). Semen from six adult male leopards was collected by electroejaculation (one ejaculation per cat)...
November 22, 2016: Reproduction in Domestic Animals, Zuchthygiene
José Rafael Blesa, Julio Tudela, Justo Aznar
: Somatic cell nuclear transfer (SCNT) (cloning), as a reproductive or therapeutic method, and mitochondrial DNA transfer, as a method to prevent the transmission of mitochondrial diseases, are analyzed in this paper from a bioethics perspective. The licit purpose of being able to treat certain diseases, as in the case of SCNT, cannot justify, in any case, resorting to illicit means such as the manipulation, selection, and elimination of human embryos in the blastocyst phase, by using cell lines obtained from them...
May 2016: Linacre Quarterly
Laura Rienzi, Clarisa Gracia, Roberta Maggiulli, Andrew R LaBarbera, Daniel J Kaser, Filippo M Ubaldi, Sheryl Vanderpoel, Catherine Racowsky
BACKGROUND: Successful cryopreservation of oocytes and embryos is essential not only to maximize the safety and efficacy of ovarian stimulation cycles in an IVF treatment, but also to enable fertility preservation. Two cryopreservation methods are routinely used: slow-freezing or vitrification. Slow-freezing allows for freezing to occur at a sufficiently slow rate to permit adequate cellular dehydration while minimizing intracellular ice formation. Vitrification allows the solidification of the cell(s) and of the extracellular milieu into a glass-like state without the formation of ice...
November 8, 2016: Human Reproduction Update
Atsushi Fukuda, Atsushi Mitani, Toshiyuki Miyashita, Takashi Sado, Akihiro Umezawa, Hidenori Akutsu
In female mammals, activation of Xist (X-inactive specific transcript) is essential for establishment of X chromosome inactivation. During early embryonic development in mice, paternal Xist is preferentially expressed whereas maternal Xist (Xm-Xist) is silenced. Unlike autosomal imprinted genes, Xist imprinting for Xm-Xist silencing was erased in cloned or parthenogenetic but not fertilized embryos. However, the molecular mechanism underlying the variable nature of Xm-Xist imprinting is poorly understood. Here, we revealed that Xm-Xist silencing depends on chromatin condensation states at the Xist/Tsix genomic region and on Rnf12 expression levels...
October 2016: PLoS Genetics
Takanobu Inoue, Atsuo Iida, Shingo Maegawa, Atsuko Sehara-Fujisawa, Masato Kinoshita
In this study, we verified nuclear transport activity of an artificial nuclear localization signal (aNLS) in medaka fish (Oryzias latipes). We generated a transgenic medaka strain expresses the aNLS tagged enhanced green fluorescent protein (EGFP) driven by a medaka beta-actin promoter. The aNLS-EGFP was accumulated in the nuclei of somatic tissues and yolk nuclei of oocytes, but undetectable in the spermatozoa. The fluorescent signal was observed from immediately after fertilization by a maternal contribution...
October 19, 2016: Development, Growth & Differentiation
Long Jin, Hai-Ying Zhu, Qing Guo, Xiao-Chen Li, Yu-Chen Zhang, Cheng-Du Cui, Wen-Xue Li, Zheng-Yun Cui, Xi-Jun Yin, Jin-Dan Kang
Cloning remains as an important technique to enhance the reconstitution and distribution of animal population with high-genetic merit. One of the major detrimental factors of this technique is the abnormal epigenetic modifications. MGCD0103 is known as a histone deacetylase inhibitor. In this study, we investigated the effect of MGCD0103 on the in vitro blastocyst formation rate in porcine somatic cell nuclear transferred (SCNT) embryos and expression in acetylation of the histone H3 lysine 9 and histone H4 lysine 12...
September 13, 2016: Theriogenology
Valerie C Coffman, Matthew B A McDermott, Blerta Shtylla, Adriana T Dawes
Positioning of microtubule organizing centers (MTOCs) incorporates biochemical and mechanical cues for proper alignment of the mitotic spindle and cell division site. Current experimental and theoretical studies in the early C. elegans embryo assume remarkable changes in the origin and polarity of forces acting on the MTOCs. These changes must occur over a few minutes, between initial centration and rotation of the pronuclear complex and entry into mitosis, and the models do not replicate in vivo timing of centration and rotation...
October 12, 2016: Molecular Biology of the Cell
Bjoern Petersen, Antje Frenzel, Andrea Lucas-Hahn, Doris Herrmann, Petra Hassel, Sabine Klein, Maren Ziegler, Klaus-Gerd Hadeler, Heiner Niemann
BACKGROUND: Xenotransplantation is considered to be a promising solution to the growing demand for suitable donor organs for transplantation. Despite tremendous progress in the generation of pigs with multiple genetic modifications thought to be necessary to overcoming the severe rejection responses after pig-to-non-human primate xenotransplantation, the production of knockout pigs by somatic cell nuclear transfer (SCNT) is still an inefficient process. Producing genetically modified pigs by intracytoplasmic microinjection of porcine zygotes is an alluring alternative...
September 2016: Xenotransplantation
Maria Giulia Minasi, Alessandro Colasante, Teresa Riccio, Alessandra Ruberti, Valentina Casciani, Filomena Scarselli, Francesca Spinella, Francesco Fiorentino, Maria Teresa Varricchio, Ermanno Greco
STUDY QUESTION: Are there correlations among human blastocyst ploidy status, standard morphology evaluation and time-lapse kinetics? SUMMARY ANSWER: Correlations were observed, in that euploid human blastocysts showed a higher percentage with top quality inner cell mass (ICM) and trophectoderm (TE), higher expansion grades and shorter time to start of blastulation, expansion and hatching, compared to aneuploid ones. WHAT IS KNOWN ALREADY: Embryo quality has always been considered an important predictor of successful implantation and pregnancy...
October 2016: Human Reproduction
Xiaoxiao Hou, Jun Liu, Zhiren Zhang, Yanhui Zhai, Yutian Wang, Zhengzhu Wang, Bo Tang, Xueming Zhang, Liguang Sun, Ziyi Li
DNA methylation and histone modification play important roles in the development of mammalian embryos. Cytochalasin B (CB) is an actin polymerization inhibitor that can significantly affect cell activity and is often used in studies concerning cytology. In recent years, CB is also commonly being used in in vitro experiments on mammalian embryos, but few studies have addressed the effect of CB on the epigenetic modification of embryonic development, and the mechanism underlying this process is also unknown. This study was conducted to investigate the effects of CB on DNA methylation and histone modification in the development of parthenogenetically activated porcine embryos...
November 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
John Zhang, Guanglun Zhuang, Yong Zeng, Jamie Grifo, Carlo Acosta, Yimin Shu, Hui Liu
Nuclear transfer of an oocyte into the cytoplasm of another enucleated oocyte has shown that embryogenesis and implantation are influenced by cytoplasmic factors. We report a case of a 30-year-old nulligravida woman who had two failed IVF cycles characterized by all her embryos arresting at the two-cell stage and ultimately had pronuclear transfer using donor oocytes. After her third IVF cycle, eight out of 12 patient oocytes and 12 out of 15 donor oocytes were fertilized. The patient's pronuclei were transferred subzonally into an enucleated donor cytoplasm resulting in seven reconstructed zygotes...
October 2016: Reproductive Biomedicine Online
Tayita Suttirojpattana, Tamas Somfai, Satoko Matoba, Takashi Nagai, Rangsun Parnpai, Masaya Geshi
We assessed the effect of pretreating sperm with dithiobutylamine (DTBA) to improve embryo development by intracytoplasmic sperm injection (ICSI) in cows. Acridine Orange staining revealed that when applied at different concentrations (2.5, 5, and 10 mM) and exposure times (5 min, 20 min, 1 h, and 2 h), DTBA reduced disulfide bonds in spermatozoa with the highest efficacy at 5 mM for 5 min. DTBA enhanced the percentage of spermatozoa with free protamine thiol groups compared with untreated spermatozoa (control) (P < 0...
August 14, 2016: Journal of Reproduction and Development
Manita Wittayarat, Yoko Sato, Lanh Thi Kim Do, Kaywalee Chatdarong, Theerawat Tharasanit, Mongkol Techakumphu, Masayasu Taniguchi, Takeshige Otoi
This study aimed to determine the acetylation at lysine 9/18/23 of histone H3 (H3K9ac/H3K18ac/H3K23ac; H3K9/18/23 ac) and the di-methylation at lysine 9 of histone H3 (H3K9me2) during early embryogenesis among trichostatin A (TSA)-treated interspecies somatic cell nuclear transfer (iSCNT) cat-cow (TSA-iSCNT) embryos, TSA-untreated iSCNT cat-cow control (control) embryos and bovine in vitro fertilization (IVF) embryos, because TSA-iSCNT embryos can develop to blastocysts. Compared to the control embryos, higher expressions of H3K9/18/23 ac were observed in TSA-iSCNT embryos and IVF embryos at most following stages (2 h post-fusion / post-insemination (PF/PI) to eight-cell stage)...
August 9, 2016: Animal Science Journal, Nihon Chikusan Gakkaihō
Jiaqiang Wang, Xin Li, Leyun Wang, Jingyu Li, Yanhua Zhao, Gerelchimeg Bou, Yufei Li, Guanyi Jiao, Xinghui Shen, Renyue Wei, Shichao Liu, Bingteng Xie, Lei Lei, Wei Li, Qi Zhou, Zhonghua Liu
Endogenous retroviruses (ERVs) are transcriptionally active in cleavage stage embryos, yet their functions are unknown. ERV sequences are present in the majority of long intergenic noncoding RNAs (lincRNAs) in mouse and humans, playing key roles in many cellular processes and diseases. Here, we identify LincGET as a nuclear lincRNA that is GLN-, MERVL-, and ERVK-associated and essential for mouse embryonic development beyond the two-cell stage. LincGET is expressed in late two- to four-cell mouse embryos. Its depletion leads to developmental arrest at the late G2 phase of the two-cell stage and to MAPK signaling pathway inhibition...
October 2016: EMBO Reports
Ying Liang, Qian-hua Tian, Yu-xia Mu, Hui-lan Du
OBJECTIVE: To observe the effect of Cangfu Congxian Decoction (CCD) on oxidative stress in granulosa cells of polycystic ovary syndrome (PCOS) patients. METHODS: Forty PCOS patients underwent in vitro fertilization-embryo transfer (IVF-ET) were assigned to the treatment group and the control group 1 according to random digit table, 20 in each group. Patients in the treatment group took CCD (200 mL, once in the morning and once in the afternoon) 2 months before IVF-ET, while those in the control group 1 took no Chinese medical decoction...
June 2016: Chinese Journal of Integrated Traditional and Western Medicine
Jochen Tigges, Erhard Godehardt, Therese Soepenberg, Beatrice Maxrath, Kerstin Friol, Christian Gnoth
PURPOSE: To analyze cumulative and single-cycle success rates of IVF and intracytoplasmic sperm injection (ICSI) separately in relation to female age, fertilization modality, and first-cycle outcome. METHODS: The study involved 2997 patients and couples, respectively, who underwent 5339 fresh and 3006 cryo cycles using pronuclear-stage oocytes. Fertilization was achieved by the conventional IVF or ICSI. Because of legal restrictions in Germany, no embryo selection was performed...
November 2016: Archives of Gynecology and Obstetrics
Reza K Oqani, Tao Lin, Jae Eun Lee, So Yeon Kim, Soo Jin Sa, Je Seok Woo, Dong Il Jin
Positive transcription elongation factor b (P-TEFb) is an RNA polymerase II kinase that phosphorylates Ser2 of the carboxyl-terminal domain and promotes the elongation phase of transcription. Despite the fact that P-TEFb has role in many cellular processes, the role of this kinase complex remains to be understood in early developmental events. In this study, using immunocytochemical analyses, we find that the P-TEFb components, Cyclin T1, CDK9, and its T-loop phosphorylated form, are localized to nuclear speckles, as well as in nucleoli in mouse germinal vesicle oocytes...
September 2016: Genesis: the Journal of Genetics and Development
Xiaonan Qi, Huatao Li, Xia Cong, Xin Wang, Zhongling Jiang, Rongfeng Cao, Wenru Tian
Scutellaria baicalensis has been effectively used in Chinese traditional medicine to prevent miscarriages. However, little information is available on its mechanism of action. This study is designed specifically to reveal how baicalin, the main effective ingredient of S. baicalensis, improves developmental competence of embryos in vitro, using the mouse as a model. Mouse pronuclear embryos were cultured in KSOM medium supplemented with (0, 2, 4 and 8 μg/ml) baicalin. The results demonstrated that in vitro culture conditions significantly decreased the blastocyst developmental rate and blastocyst quality, possibly due to increased cellular stress and apoptosis...
July 29, 2016: Journal of Reproduction and Development
Masakazu Hashimoto, Yukiko Yamashita, Tatsuya Takemoto
The CRISPR/Cas9 system is a powerful tool for elucidating the roles of genes in a wide variety of organisms including mice. To obtain genetically modified embryos or mice by this method, Cas9 mRNA and sgRNA are usually introduced into zygotes by microinjection or electroporation. However, most mutants generated with this method are genetically mosaic, composed of several types of cells carrying different mutations, which complicates phenotype analysis in founder embryos or mice. To simplify the analysis and to elucidate the roles of genes involved in developmental processes, a method for producing non-mosaic mutants is needed...
October 1, 2016: Developmental Biology
Yan Li, Di Lian, Shoulong Deng, Xiaosheng Zhang, Jinlong Zhang, Wenting Li, Hai Bai, Zhixian Wang, Hongping Wu, Juncai Fu, Hongbing Han, Jianzhong Feng, Guoshi Liu, Ling Lian, Zhengxing Lian
BACKGROUND: Brucella is a zoonotic Gram-negative pathogen that causes abortion and infertility in ruminants and humans. TLR4 is the receptor for LPS which can recognize Brucella and initiate antigen-presenting cell activities that affect both innate and adaptive immunity. Consequently, transgenic sheep over-expressing TLR4 are an suitable model to investigate the effects of TLR4 on preventing Brucellosis. In this study, we generated transgenic sheep overexpressing TLR4 and aimed to evaluate the effects of different seasons (breeding and non-breeding season) on superovulation and the imported exogenous gene on growth...
2016: Journal of Animal Science and Biotechnology
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