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zygote activation

Bo Fu, Hong Ma, Di Liu
Pre-implantation embryo development encompasses several key developmental events, especially the activation of zygotic genome activation (ZGA)-related genes. Endogenous retroviruses (ERVs), which are regarded as "deleterious genomic parasites", were previously considered to be "junk DNA". However, it is now known that ERVs, with limited conservatism across species, mediate conserved developmental processes (e.g., ZGA). Transcriptional activation of ERVs occurs during the transition from maternal control to zygotic genome control, signifying ZGA...
February 12, 2019: International Journal of Molecular Sciences
Nan He, Bradley H Rosen, Jaimie S Gray, Idil A Evans, Marina Zieger, Ziying Yan, Florie Borel, Bo Liang, Xingshen Sun, Shashanna R Moll, Michael H Brodsky, Christian Mueller, John F Engelhardt
RATIONALE: The most prevalent genetic cause of chronic obstructive pulmonary disease is alpha-1 antitrypsin (A1AT) deficiency, a disorder that has yet to be widely modeled in animals because of species-specific differences between rodents and humans. OBJECTIVES: To address these challenges, we engineered two A1AT ferret models using zygote gene editing to test the hypothesis that unopposed protease activity within the lung leads to emphysema and bronchitis. METHODS: Guide RNAs targeting exon 2 (for knockout) and exon 5 (for Z-allele mutation, Pi*Z) of the ferret A1AT gene were injected into ferret zygotes with Cas9 mRNA...
December 2018: Annals of the American Thoracic Society
Gerald Lincoln
The tenet of this review is that innate circannual timing is an ancestral trait that first evolved in free-living eukaryotic cells some 2000My ago. Marine algae of the genus Allexandrium provide a living unicellular model. This species shows the primitive trait of 'alternation of generations' where the organism alternates between fast replicating vegetative cells in summer, and a dormant cystic cell over winter. The resistant cysts sink into the cold ocean sediments. Remarkably, excystment in spring is governed by an endogenous circannual timing mechanism...
February 9, 2019: Journal of Neuroendocrinology
Hye Kyung Lee, Michaela Willi, Harold E Smith, Shannon M Miller, David R Liu, Chengyu Liu, Lothar Hennighausen
A particular challenge in genome engineering has been the simultaneous introduction of mutations into linked (located on the same chromosome) loci. Although CRISPR/Cas9 has been widely used to mutate individual sites, its application in simultaneously targeting of linked loci is limited as multiple nearby double-stranded DNA breaks created by Cas9 routinely result in the deletion of sequences between the cleavage sites. Base editing is a newer form of genome editing that directly converts C∙G-to-T∙A, or A∙T-to-G∙C, base pairs without introducing double-stranded breaks, thus opening the possibility to generate linked mutations without disrupting the entire locus...
February 7, 2019: Scientific Reports
Qian-Qian Sha, Jue Zhang, Heng-Yu Fan
In mammals, maternal-to-zygotic transition (MZT), or oocyte-to-embryo transition, begins with oocyte meiotic resumption due to the sequential translational activation and destabilization of dormant maternal transcripts stored in the ooplasm. It then continues with the elimination of maternal transcripts during oocyte maturation and fertilization and ends with the full transcriptional activation of the zygotic genome during embryonic development. A hallmark of MZT in mammals is its reliance on translation and the utilization of stored RNAs and proteins, rather than de novo transcription of genes, to sustain meiotic maturation and early development...
January 31, 2019: Biology of Reproduction
Yue Liu, Songhua Xu, Xiuli Lian, Yang Su, Yuhuan Zhong, Ruimin Lv, Kaien Mo, Huimin Zhu, Xiaojiang Wang, Lixuan Xu, Shie Wang
Zygotic genome activation (ZGA) is one of the most critical events at the beginning of mammalian preimplantation embryo development (PED). The mechanisms underlying mouse ZGA remain unclear although it has been widely studied. In the present study, we identified that tricho-rhino-phalangeal syndrome 1 (TRPS1), an atypical GATA family member, is an important factor for ZGA in mouse PED. We found that the Trps1 mRNA level peaked at the one-cell stage while TRPS1 protein did so at the two/four-cell stage. Knockdown of Trps1 by the microinjection of Trps1 siRNA reduced the developmental rate of mouse preimplantation embryos by approximately 30%, and increased the expression of ZGA marker genes MuERV-L and Zscan4d via suppressing the expression of major histone markers H3K4me3 and H3K27me3...
February 2, 2019: Cell Cycle
Tingting Sui, Di Liu, Tingjun Liu, Jichao Deng, Mao Chen, Yuanyuan Xu, Yuning Song, Hongsheng Ouyang, Liangxue Lai, Zhanjun Li
Premature aging syndromes are rare genetic disorders mimicking clinical and molecular features of aging. Products of the LMNA gene, primarily lamin A and C, are major components of the nuclear lamina. A recently identified group of premature aging syndromes was related to mutations of the LMNA gene. Although LMNA disorders have been identified in premature aging syndromes, affect specifically the skeletal muscles, cardiac muscles, and lipodystrophy, understanding the pathogenic mechanisms still need to be elucidated...
February 2019: Aging and Disease
Chien-Hsun Huang, Fu-Ting Wang, Wen-Hsiung Chan
Ochratoxin A (OTA), a mycotoxin constituent of a range of food commodities, including coffee, wine, beer, grains, and spices, exerts toxicological and pathological effects in vivo, such as nephrotoxicity, hepatotoxicity, and immunotoxicity. In a previous report, we highlighted the potential of OTA to induce apoptosis via reactive oxygen species (ROS) generation in mouse blastocysts that led to impaired preimplantation and postimplantation embryo development in vitro and in vivo. Here, we have shown that liquiritigenin (LQ), a type of flavonoid isolated from Glycyrrhiza radix, effectively protects against OTA-mediated apoptosis and inhibition of cell proliferation in mouse blastocysts...
January 30, 2019: Environmental Toxicology
Mélanie Eckersley-Maslin, Celia Alda-Catalinas, Marloes Blotenburg, Elisa Kreibich, Christel Krueger, Wolf Reik
The molecular regulation of zygotic genome activation (ZGA) in mammals remains an exciting area of research. Primed mouse embryonic stem cells contain a rare subset of "2C-like" cells that are epigenetically and transcriptionally similar to the two-cell embryo and thus represent an in vitro approximation for studying ZGA transcription regulation. Recently, the transcription factor Dux, expressed in the minor wave of ZGA, was described to activate many downstream ZGA transcripts. However, it remains unknown what upstream maternal factors initiate ZGA in either a Dux-dependent or Dux-independent manner...
January 28, 2019: Genes & Development
Runshuai Zhang, Kongyue Wu, Xiaomei Ke, Xueyan Zhang, Gongyu Xu, Hao Shen, Emile Nibona, Abdullah Al Hafiz, Xiaoting Liang, Zequn Wang, Chao Qi, Qingchun Zhou, Xueping Zhong, Haobin Zhao
B-cell lymphoma-6 (Bcl6) is a transcriptional repressor that plays important roles in various physiological activities such as innate and adaptive immune response, lymphocyte differentiation, and cell cycle regulation in mammals. Two homologs of Bcl6a, namely Bcl6aa and Bcl6ab, are identified in teleost fish including medaka Oryzias latipes. The expression profiles of bcl6aa and bcl6ab in medaka were studied using reverse-transcription polymerase chain reaction and in situ hybridization. The transcripts of bcl6aa and bcl6ab were detected from very early embryos such as the four-cell stage until hatching...
January 25, 2019: Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution
Marina Veil, Lev Yampolsky, Bjoern Gruening, Daria Onichtchouk
The zebrafish embryo is mostly transcriptionally quiescent during the first 10 cell cycles, until the main wave of Zygotic Genome Activation (ZGA) occurs, accompanied by fast chromatin remodeling. At ZGA, homologs of mammalian stem cell transcription factors (TFs) Pou5f3, Nanog and Sox19b bind to thousands of developmental enhancers to initiate transcription. So far, how these TFs influence chromatin dynamics at ZGA has remained unresolved. To address this question, we analyzed nucleosome positions in wild-type and Maternal-Zygotic (MZ) mutants for pou5f3 and nanog by MNase-seq...
January 23, 2019: Genome Research
Huili Wang, Erika E Paulson, Libing Ma, Pablo J Ross, Richard M Schultz
Enhancer of zeste homolog 2 (EZH2), a component of the PRC2 complex, trimethylates H3K27, a transcriptionally repressive histone mark. EZH2 is encoded by a dormant maternal mRNA and inhibiting the maturation-associated increase in EZH2 activity using either a combined siRNA/morpholino approach or a small molecule inhibitor (GSK343) inhibits development of diploidized parthenotes to the blastocyst stage but not inseminated eggs, with longer GSK343 treatments leading to progressively greater inhibition of development...
January 19, 2019: Epigenetics: Official Journal of the DNA Methylation Society
Nami Morishita, Masanori Ochi, Toshitaka Horiuchi
Purpose: To investigate the effects of sperm treatment medium-TCM199 or EGTA in Tris-HCl buffer (TBS + EGTA)-for sonication of frozen-thawed hamster spermatozoa in terms of sperm chromosome integrity and development of hamster oocytes injected with the sperm heads (ICSI). Methods: Frozen-thawed hamster spermatozoa were separated into heads and tails by sonication in TCM199 or TBS + EGTA. Sperm heads were injected into mouse oocytes to assess hamster sperm chromosomes...
January 2019: Reproductive Medicine and Biology
Chu Chu, Zhaohui Yang, Jiayin Yang, Li Yan, Chenyang Si, Yu Kang, Zhenzhen Chen, Yongchang Chen, Weizhi Ji, Yuyu Niu
BACKGROUND: Non-human primate (NHP) models can closely mimic human physiological functions and are therefore highly valuable in biomedical research. Genome editing is now developing rapidly due to the precision and efficiency offered by engineered site-specific endonuclease-based systems, such as transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) system. It has been demonstrated that these programmable nucleases can introduce genetic changes in embryos from many species including NHPs...
January 15, 2019: BMC Biotechnology
April DeLaurier, Cynthia Lizzet Alvarez, Kali J Wiggins
Background: Histone deacetylases (HDACs) are epigenetic factors that function to repress gene transcription by removing acetyl groups from the N-terminal of histone lysines. Histone deacetylase 4 (HDAC4), a class IIa HDAC, has previously been shown to regulate the process of endochondral ossification in mice via repression of Myocyte enhancer factor 2c (MEF2C), a transcriptional activator of Runx2 , which in turn promotes chondrocyte maturation and production of bone by osteoblasts. Methods & Materials: In this study, we generated two zebrafish lines with mutations in hdac4 using CRISPR/Cas9 and analyzed mutants for skeletal phenotypes and expression of genes known to be affected by Hdac4 expression...
2019: PeerJ
Yuki Shindo, Amanda A Amodeo
During zygotic genome activation (ZGA), the chromatin environment undergoes profound changes, including the formation of topologically associated domains, refinements in nucleosome positioning on promoters, and the emergence of heterochromatin [1-4]. In many organisms, including Drosophila, ZGA is associated with the end of a period of extremely rapid, exponential cleavage divisions that are facilitated by large maternally provided pools of nuclear components. It is therefore imperative that we understand how the supply of chromatin components relative to the exponentially increasing demand affects nuclear and chromatin composition during early embryogenesis...
January 8, 2019: Current Biology: CB
Muhammad Assad Aslam, Mir Farshid Alemdehy, Colin E J Pritchard, Ji-Ying Song, Fitriari Izzatunnisa Muhaimin, Ruud H Wijdeven, Ivo J Huijbers, Jacques Neefjes, Heinz Jacobs
C9orf82 protein, or conserved anti-apoptotic protein 1 or caspase activity and apoptosis inhibitor 1 (CAAP1) has been implicated as a negative regulator of the intrinsic apoptosis pathway by modulating caspase expression and activity. In contrast, an independent genome wide screen for factors capable of driving drug resistance to the topoisomerase II (Topo II) poisons doxorubicin and etoposide, implicated a role for the nuclear protein C9orf82 in delaying DSBs repair downstream of Topo II, hereby sensitizing cells to DSB induced apoptosis...
2019: PloS One
Anna-Rita Piras, Irene Menéndez-Blanco, Sandra Soto-Heras, Maria-Gracia Catalá, Dolors Izquierdo, Luisa Bogliolo, Maria-Teresa Paramio
This study aimed to investigate the effect of resveratrol supplementation in maturation medium on the developmental ability and bioenergetic\oxidative status of prepubertal goat oocytes selected by brilliant cresyl blue (BCB). Oocytes collected from slaughterhouse-derived ovaries were selected by 13 µM BCB staining and classified as grown BCB+ and growing BCB- oocytes. All oocytes were matured in vitro in our conventional maturation medium and supplemented with 1 µM (BCB+R and BCB-R) and without (Control groups: BCB+C and BCB-C) resveratrol...
December 29, 2018: Journal of Reproduction and Development
Renato Pereira da Costa Gerger, Eduardo de Souza Ribeiro, Fabiano Carminatti Zago, Luís Henrique de Aguiar, Paula Rodriguez-Villamil, Felipe Ledur Ongaratto, Carlos Eduardo Ambrósio, Maria Angélica Miglino, José Luiz Rodrigues, Fabiana Forell, Luciana Relly Bertolini, Marcelo Bertolini
Nuclear reprogramming in somatic cell cloning is one of the key factors for proper development, with variations in the protocol appearing to improve cloning efficiency. This study aimed to determine the effects of two fusion-activation intervals and the aggregation of bovine cloned embryos on subsequent in vitro and in vivo development. Zygotes produced by handmade cloning were exposed to two fusion-activation intervals (2 h or 4 h), and then cultured in microwells either individually (1 × 100%) or after aggregation of two structures (2 × 100%)...
December 15, 2018: Research in Veterinary Science
Federica Marinaro, Beatriz Macías-García, Francisco Miguel Sánchez-Margallo, Rebeca Blázquez, Verónica Álvarez, Elvira Matilla, Nuria Hernández, María Gómez-Serrano, Inmaculada Jorge, Jesús Vázquez, Lauro González-Fernández, Eva Pericuesta, Alfonso Gutiérrez-Adán, Javier G Casado
Advanced age is a risk factor undermining women´s fertility. Hence, the optimization of assisted reproduction techniques is an interdisciplinary challenge that requires the improvement of in vitro culture systems. Here, we hypothesize that supplementation of embryo culture medium with extracellular vesicles from endometrial-derived mesenchymal stem cells (EV-endMSCs) may have a positive impact on the embryo competence of aged oocytes. In this work, 24 weeks old B6D2 female mice were used as egg donors and in vitro fertilization assays were performed using males from the same strain (8-12 weeks); the presumptive zygotes were incubated in the presence of 0, 10, 20, 40 or 80 μg/ml of EV-endMSCs...
December 28, 2018: Biology of Reproduction
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