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1.
Small ; 19(22): e2207966, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36861366

RESUMO

Herein, a novel combination of Mg- and Ga-co-doped ZnO (MGZO)/Li-doped graphene oxide (LGO) transparent electrode (TE)/electron-transporting layer (ETL) has been applied for the first time in Cu2 ZnSn(S,Se)4 (CZTSSe) thin-film solar cells (TFSCs). MGZO has a wide optical spectrum with high transmittance compared to that with conventional Al-doped ZnO (AZO), enabling additional photon harvesting, and has a low electrical resistance that increases electron collection rate. These excellent optoelectronic properties significantly improved the short-circuit current density and fill factor of the TFSCs. Additionally, the solution-processable alternative LGO ETL prevented plasma-induced damage to chemical bath deposited cadmium sulfide (CdS) buffer, thereby enabling the maintenance of high-quality junctions using a thin CdS buffer layer (≈30 nm). Interfacial engineering with LGO improved the Voc of the CZTSSe TFSCs from 466 to 502 mV. Furthermore, the tunable work function obtained through Li doping generated a more favorable band offset in CdS/LGO/MGZO interfaces, thereby, improving the electron collection. The MGZO/LGO TE/ETL combination achieved a power conversion efficiency of 10.67%, which is considerably higher than that of conventional AZO/intrinsic ZnO (8.33%).

2.
J Reprod Dev ; 66(3): 255-263, 2020 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-32213735

RESUMO

The large Japanese field mouse (Apodemus speciosus) is endemic to Japan and may be used as an animal model for studies related to environmental pollution, medical science, and basic biology. However, the large Japanese field mouse has low reproductive ability due to the small number of oocytes ovulated per female. To produce experimental models, we investigated the in vitro developmental potential of interspecies somatic cell nuclear transfer (iSCNT) embryos produced by fusing tail tip cells from the large Japanese field mouse with enucleated oocytes from laboratory mice (Mus musculus domesticus). Only a small number of iSCNT embryos developed to the 4-cell (0-4%) and blastocysts (0-1%) stages under sequential treatment using trichostatin A (TSA) and vitamin C (VC) supplemented with deionized bovine serum albumin (d-BSA). This sequential treatment led to the reduction in H3K9 trimethylation and did not affect H3K4 trimethylation in at least the 2-cell stage of the iSCNT embryos. Moreover, iSCNT embryos that received tail tip cells with exposure treatment to ooplasm from cell fusion to oocyte activation or VC treatment prior to cell fusion did not exhibit significant in vitro development improvement compared to that of each control group. This suggests that large Japanese field mice/laboratory mice iSCNT embryos that received sequential treatment using TSA and VC with d-BSA may have slightly better developmental potential beyond the 4-cell stage. Our results provide insights into the reprogramming barriers impeding the wider implementation of iSCNT technology.


Assuntos
Clonagem de Organismos/métodos , Desenvolvimento Embrionário/fisiologia , Técnicas de Transferência Nuclear , Oócitos/citologia , Animais , Embrião de Mamíferos/citologia , Feminino , Camundongos , Murinae
3.
Small ; 14(30): e1801226, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29966042

RESUMO

Herein, bimetallic iron (Fe)-manganese (Mn) oxyhydroxide ((Fe1-x, Mnx )OOH, FeMnOOH) nanosheets on fluorine-doped tin oxide conducting substrates and on semiconductor photoanodes are synthesized by a facile, room temperature, electroless deposition method as catalysts for both electrochemical and photo-electrochemical (PEC) water splitting, respectively. Surprisingly, Mn-doped FeOOH can significantly modulate the nanosheet morphology to increase the active surface area, boost more active sites, and augment the intrinsic activity by tuning the electronic structure of FeOOH. Due to the 2D nanosheet architecture, the optimized FeMnOOH exhibits superior electrochemical activity and outstanding durability for the oxygen evolution reaction with a low overpotential of 246 mV at 10 mA cm-2 and 414 mV at 100 mA cm-2 , and long-term stability for 40 h without decay, which is comparable to the best electrocatalysts for water oxidation reported in the literature. By integrating with semiconductor photoanodes (such as α-Fe2 O3 nanorod (NR) arrays), bimetallic FeMnOOH catalysts achieve solar-driven water splitting with a significantly enhanced PEC performance (3.36 mA cm-2 at 1.23 V vs reversible hydrogen electrode (RHE)) with outstanding long-term stability (≈8 h) compared to that of the bare Fe2 O3 NR (0.92 mA cm-2 at 1.23 V vs RHE).

4.
J Reprod Dev ; 64(1): 65-74, 2018 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-29212961

RESUMO

Maternal RNA/protein degradation and zygotic genome activation (ZGA), occurring during maternal-to-zygotic transition (MZT), are the first essential events for the development of pre-implantation embryos. Previously, we have shown the importance of the ubiquitin-proteasome system (UPS) for initiation of minor ZGA at the 1-cell stage of mouse embryos. However, little is known about the mechanism of involvement of the UPS-degraded maternal proteins in ZGA. In this study, we investigated the effect of inhibiting maternal protein degradation by the reversible proteasome inhibitor, MG132, on post-implantation development and ZGA regulation during early cleavage stages. Our study revealed that zygotic transcription by RNA polymerase II (Pol II) at the 1-cell stage was delayed and the full-term development was affected by transient proteasome inhibition during 1 to 9 h post-insemination (hpi). Furthermore, we found that the transient inhibition of proteasome activity at the 2-cell stage delayed the onset of transcription of some major ZGA genes. These results support the model hypothesizing the requirement of sequential degradation of maternal proteins by UPS for the proper onset of ZGA and normal progression of MZT in early mouse embryos.


Assuntos
Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Complexo de Endopeptidases do Proteassoma/genética , Ubiquitina/genética , Animais , Camundongos , Oócitos/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Ubiquitina/metabolismo
5.
Phys Chem Chem Phys ; 17(30): 19777-88, 2015 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-26153341

RESUMO

A quaternary indium- and gallium-free kesterite (KS)-based compound, copper zinc tin sulfide (Cu2ZnSnS4, CZTS), has received significant attention for its potential applications in low cost and sustainable solar cells. It is well known that the reaction time, reactivity of the precursors, and types of capping ligands used during the synthesis of colloidal nanocrystals (NCs) strongly influence the crystallographic phase of the NCs. In this research, a non-toxic and green synthetic strategy for both the synthesis of CZTS NCs and the fabrication of a highly efficient CZTS absorber layers using an ink formulation without a toxic solvent, which meets the comprehensive framework for green chemistry that covers major aspects of the environmental strain, is demonstrated. In particular, pure metastable wurtzite (WZ) CZTS NCs are synthesized using the environmentally harmless, polyol mediated hot-injection (HI) technique at a low reaction temperature. The influence of the reaction time on the properties of the CZTS NCs is investigated in detail. Based on detailed reaction time dependent phase evolution, a possible growth and formation mechanism is proposed. Furthermore, a scalable, low cost, binder free ink formulation process without ligand exchange is developed using ethanol as the dispersal solvent. The as-prepared WZ-derived CZTS NC thin films are observed to undergo a phase transformation to KS during annealing in a sulfur vapor atmosphere via rapid thermal annealing above 500 °C, and surprisingly, this process results in fully sintered, compact and uniform CZTS thin films with large sized grains. The best solar cell device fabricated using a CZTS absorber that was sulfurized at an optimized temperature exhibits a power conversion efficiency of 2.44%, which is the highest efficiency obtained using the polyol-based HI route.

6.
J Reprod Dev ; 60(3): 179-86, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24583807

RESUMO

In the mammalian testis, the ubiquitin-proteasome system plays important roles in the process that promotes the formation of mature sperm. We recently identified zygote-specific proteasome assembly chaperone (ZPAC), which is specifically expressed in the mouse gonads and zygote. ZPAC mediates a unique proteasome assembly pathway in the zygote, but the expression profile and function of ZPAC in the testis is not fully understood. In this study, we investigated the possible role of ZPAC during mouse spermatogenesis. First, we analyzed the expression of ZPAC and 20S proteasome subunit α4/PSMA7 in the adult mouse testis. ZPAC and α4 were expressed in spermatogonia, spermatocytes, and round spermatids. In elongating spermatids, ZPAC was expressed until step 10, whereas expression of α4 persisted until step 12. We then examined the expression profile of ZPAC and α4 in a mouse model of experimental unilateral cryptorchidism. Consistent with appearance of morphologically impaired germ cells following cryptorchidism, the ZPAC protein level was significantly decreased at 4 days post induction of experimental cryptorchidism (D4) compared with the intact testis, although the amount of α4 protein persisted at least until D10. Moreover, intense ZPAC staining was co-localized with staining of annexin V, an early indicator of apoptosis in mammalian cells, in germ cells of cryptorchid testis, but ZPAC was also expressed in germ cells showing no detectable expression of annexin V. These results suggest that ZPAC plays a role during spermatogenesis and raises the possibility that 20S proteasome mediated by ZPAC may be involved in the regulation of germ cell survival during spermatogenesis.


Assuntos
Chaperonas Moleculares/fisiologia , Proteínas Nucleares/fisiologia , Espermatogênese/genética , Animais , Sobrevivência Celular/genética , Criptorquidismo/genética , Criptorquidismo/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Camundongos , Camundongos Endogâmicos ICR , Chaperonas Moleculares/genética , Proteínas Nucleares/genética , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Espermatozoides/fisiologia , Testículo/metabolismo , Zigoto/metabolismo
7.
ACS Appl Mater Interfaces ; 16(13): 16328-16339, 2024 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-38516946

RESUMO

Kesterite-based Cu2ZnSn(S,Se)4 (CZTSSe) thin-film solar cells (TFSCs) are a promising candidate for low-cost, clean energy production owing to their environmental friendliness and the earth-abundant nature of their constituents. However, the advancement of kesterite TFSCs has been impeded by abundant defects and poor microstructure, limiting their performance potential. In this study, we present efficient Ag-alloyed CZTSSe TFSCs enabled by a facile metallic precursor engineering approach. The positioning of the Ag nanolayer in the metallic stacked precursor proves crucial in expediting the formation of Cu-Sn metal alloys during the alloying process. Specifically, Ag-included metallic precursors promote the growth of larger grains and a denser microstructure in CZTSSe thin films compared to those without Ag. Moreover, the improved uniformity of Ag, facilitated by the evaporation deposition technique, significantly suppresses the formation of detrimental defects and related defect clusters. This suppression effectively reduces nonradiative recombination, resulting in enhanced performance in kesterite TFSCs. This study not only introduces a metallic precursor engineering strategy for efficient kesterite-based TFSCs but also accelerates the development of microstructure evolution from metallic stacked precursors to metal chalcogenide compounds.

8.
Nanotechnology ; 24(9): 095706, 2013 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-23396187

RESUMO

Cu(2)ZnSnS(4) (CZTS) thin films were prepared by the sequential sulfurization of a co-sputtered precursor with a multitarget (Cu, ZnS, and SnS(2)) sputtering system. In order to investigate the crystallization behaviour of the thin films, the precursors were sulfurized in a tube furnace at different temperatures for different time durations. The Raman spectra of the sulfurized thin films showed that their crystallinity gradually improved with an increase in the sulfurization temperature and duration. However, transmission electron microscopy revealed an unexpected result-the precursor thin films were not completely transformed to the CZTS phase and showed the presence of uncrystallized material when sulfurized at 250-400 °C for 60 min and at 500 °C for 30 min. Thus, the crystallization of the co-sputtered precursor thin films showed a strong dependence on the sulfurization temperature and duration. The crystallization mechanism of the precursor thin films was understood on the basis of these results and has been described in this paper. The understanding of this mechanism may improve the standard preparation method for high-quality CZTS absorber layers.

9.
J Reprod Dev ; 59(3): 258-65, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23449310

RESUMO

Degradation of maternally stored mRNAs after fertilization is an essential process for mammalian embryogenesis. Maternal mRNA degradation depending on deadenylases in mammalian early embryos has been mostly speculated, rather than directly demonstrated. Previously, we found that gene expression of nocturnin, which functions as a circadian clock-controlled deadenylase in mammalian cells, was clearly changed during the maternal-to-zygotic transition (MZT). Here, we investigated the possible role of nocturnin during mouse MZT. First, we examined the expression profile and localization of nocturnin in mouse oocytes and early embryos. The abundance of Nocturnin mRNA level was significantly decreased from the MII to 4-cell stages and slightly increased from the 8-cell to blastocyst stages, whereas the Nocturnin protein level was almost stable in all examined cells including GV and MII oocytes and early embryos. Nocturnin was localized in both the cytoplasm and the nucleus of all examined cells. We then examined the effect of loss or gain of Nocturnin function on early embryonic development. Knockdown of Nocturnin by injection of Nocturnin antisense expression vector into 1-cell embryos resulted in the delay of early embryonic development to the early blastocyst stage. Moreover, Nocturnin-overexpressed embryos by injection of Nocturnin expression vector impaired their development from the 1-cell to 2-cell or 4-cell stages. These results suggest that precise expression of nocturnin is critical to proper development of early mouse embryos. Functional analysis of nocturnin may contribute to the understanding of the possible role of the deadenylase at mouse MZT.


Assuntos
Blastocisto/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Proteínas Nucleares/fisiologia , Fatores de Transcrição/fisiologia , Zigoto/fisiologia , Animais , Blastocisto/citologia , Núcleo Celular/metabolismo , Ritmo Circadiano , Citoplasma/metabolismo , Técnicas de Cultura Embrionária , Feminino , Fertilização , Fertilização in vitro , Perfilação da Expressão Gênica , Camundongos , Camundongos Endogâmicos ICR , Oócitos/citologia , Gravidez , Prenhez , Fatores de Tempo
10.
ACS Appl Mater Interfaces ; 15(17): 21123-21133, 2023 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-37083398

RESUMO

The coupling of oxygen evolution reaction (OER) catalysts with photoanodes is a promising strategy for enhancing the photoelectrochemical (PEC) performance by passivating photoanode's surface defect states and facilitating charge transfer at the photoanode/electrolyte interface. However, a serious interface recombination issue caused by poor interface and OER catalysts coating quality often limits further performance improvement of photoanodes. Herein, a rapid Fenton-like reaction method is demonstrated to produce ultrathin amorphous Ni:FeOOH catalysts with in situ-induced oxygen vacancies (Vo) to improve the water oxidation activity and stability of BiVO4 photoanodes. The combined physical characterizations, PEC studies, and density functional theory calculations revealed that the reductive environment in a Fenton-like reaction in situ produces abundant Vo in Ni:FeOOH catalysts, which significantly improves charge separation and charge transfer efficiency of BiVO4 while also offering abundant active sites and a reduced energy barrier for OER. As a result, Ni:FeOOH-Vo catalysts yielded a more than 2-fold increased photocurrent density in the BiVO4 photoanode (from 1.54 to 4.15 mA cm-2 at 1.23 VRHE), accompanied by high stability for 5 h. This work not only highlights the significance of abundant Vo in catalysts but also provides new insights into the rational design and fabrication of efficient and stable solar water-splitting systems.

11.
Neurosci Lett ; 767: 136298, 2022 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-34673147

RESUMO

Alzheimer's disease (AD) is characterized by amyloid beta (Aß) plaques and neurofibrillary tangles. AD drug development has been limited due to the presence of the blood-brain barrier (BBB), which prevents efficient uptake of therapeutics into the brain. To solve this problem, we used trans-activator of transcription (TAT)-transducing domain and added the human serum albumin (HSA) carrier to increase the half-life of the drug within the body. In addition, we included the protein of interest for lowering Aß deposition and/or neurofibrillary tangles. We made HSA fusion protein (designated AL04) which contains Cystatin C (CysC) as core mechanism of action moiety in the construct containing tandem repeat TAT (dTAT). After purification of 80KDa AL04, we investigate the therapeutic potential of AL04 in vitro and AD mouse model Tg2576. We evaluated the permeability of AL04 through the BBB using a cell-basedhuman BBB model and show that dTAT plays a role in facilitating the delivery of 80 kDa protein. We found out that AL04 attenuates Aß-induced neurotoxicity in PC12 cells. In Tg2576 mice brain, Aß plaques were dramatically reduced in AL04 treated mice. These data suggest that BBB-crossing albumin fusion protein AL04 with CysC active moiety can be a disease modifying treatment for AD.


Assuntos
Doença de Alzheimer , Peptídeos beta-Amiloides/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Cistatina C/farmacocinética , Portadores de Fármacos/farmacocinética , Albumina Sérica Humana/farmacocinética , Animais , Barreira Hematoencefálica , Encéfalo/metabolismo , Encéfalo/patologia , Cistatina C/administração & dosagem , Portadores de Fármacos/química , Produtos do Gene tat/farmacocinética , Humanos , Camundongos , Células PC12 , Ratos , Albumina Sérica Humana/química
12.
ACS Appl Mater Interfaces ; 13(3): 3959-3968, 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33463150

RESUMO

A cation substitution in Cu2ZnSn(S,Se)4 (CZTSSe) offers a viable strategy to reduce the open-circuit voltage (Voc)-deficit by altering the characteristics of band-tail states, antisite defects, and related defect clusters. Herein, we report a facile single process, i.e., simply introducing a thin Ag layer on a metallic precursor, to effectively improve the device characteristics and performances in kesterite (Agx,Cu1-x)2ZnSn(Sy,Se1-y)4 (ACZTSSe) solar cells. Probing into the relationship between the external quantum efficiency derivative (dEQE/dλ) and device performances revealed the Voc-deficit characteristics in the ACZTSSe solar cells as a function of Cu and Ag contents. The fabricated champion ACZTSSe solar cell device showed an efficiency of 12.07% and a record low Voc-deficit of 561 mV. Thorough investigations into the mechanism underpinning the improved performance in the ACZTSSe device further revealed the improved band-tailing characteristic, effective minority carrier lifetime, and diode factors as well as reduced antisite defects and related defect clusters as compared to the CZTSSe device. This study demonstrates the feasibility of effectively suppressing antisite defects, related defect clusters, and band-tailing characteristics by simply introducing a thin Ag layer on a metallic precursor in the kesterite solar cells, which in turn effectively reduces the Voc-deficit.

13.
ACS Appl Mater Interfaces ; 13(11): 13425-13433, 2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-33706505

RESUMO

Point defect engineering in Cu2ZnSnSe4 (CZTSe) thin films is the main issue to improve its device performance. This study reveals the correlation between the reaction pathway and the point defects in the CZTSe film. The reaction pathway from a metallic precursor (Mo/Zn/Sn/Cu) to a kesterite CZTSe film is varied by changing the annealing process. The synthesized CZTSe films under different reaction pathways induce different device performances with different defect energy levels, although all CZTSe films have similar structural and optical properties (Eg ∼ 1.0 eV). The admittance spectroscopy demonstrates the correlations between point defect types (VZn, ZnSn, ZnCu, CuZn, and VCu) and the reaction pathways for the formation of CZTSe films. The different growth rates of binary selenides, such as ZnSe and/or Sn-Se phases, during the annealing process are especially strongly related to the formation of point defects, leading to the different open-circuit voltages (396-451 mV) and fill factors (51-65%). The results of this study suggest that controlling the reaction pathway is an effective approach to adjust the formation of defects in the kesterite CZTSe film as well as to fabricate high-performance solar cell devices.

14.
J Nanosci Nanotechnol ; 10(5): 3686-90, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20359028

RESUMO

The nanocrystalline Zinc Sulfide (ZnS) thin films were prepared on glass substrates by chemical bath deposition (CBD) method using aqueous solutions of zinc acetate, thiourea and tri-sodium citrate in alkaline medium at 80 degrees C. The tri-sodium citrate acts as a complexing agent. The effects of complexing agent and annealing atmosphere (95%N2 + 5%H2S) on structural, morphological and optical properties of ZnS thin films were studied using X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and optical absorption. XRD study revealed that single phase ZnS powder was formed in the solution with tri-sodium citrate, however, ZnS and ZnO mixed phase powder was formed in the solution without tri-sodium citrate. The films deposited with trisodium citrate showed ZnS with hexagonal wurtzite phase. However, annealed film in (N2 + H2S) atmosphere showed cubic (zincblende) phase. FE-SEM images show that grain size of as-deposited and annealed ZnS films are about 20 nm and 50 nm, respectively. Optical absorption study showed that the films have moderate optical transmission from 65% to 75% in the visible region and the optical band gap energy of as-deposited ZnS film is 3.91 eV and it decreases to 3.73 eV after annealing.

15.
J Reprod Dev ; 56(6): 655-63, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20814167

RESUMO

In mammalian oocytes, the ubiquitin-proteasome system (UPS) is suggested to play important roles in oocyte meiosis resumption, spindle assembly, polar body emission and pronuclear formation by regulating cyclin B1 degradation. However, little is known about the direct relationship between zygotic gene activation (ZGA) and degradation of maternal proteins. Here, we investigated the role of the UPS in the onset of ZGA in early mouse embryos. First, we found degradation of cyclin B1 protein in fertilized oocytes at 1 hpi by western blot analysis and used these oocytes throughout this study. Subsequently, we determined optimal experimental conditions for transient inhibition of proteasomal activity by specific and reversible proteasomal inhibitor MG132 in the G1 phase of the first cell cycle. Under the selected optimal conditions, we subjected transient MG132-treated embryos to reverse transcription (RT)-PCR analysis of expression of four ZGA genes, i.e., the hsp70.1, MuERV-L, eif-1a and zscan4d genes. As a result, we found that onset of expression of the four examined ZGA genes was delayed in both normally developed 2-cell embryos and arrested 1-cell embryos. Our results indicate that proteasomal degradation of proteins by the UPS plays a pivotal role in the molecular mechanisms of ZGA in early mouse embryos.


Assuntos
Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/fisiologia , Ativação Transcricional/efeitos dos fármacos , Ubiquitina/antagonistas & inibidores , Zigoto/efeitos dos fármacos , Animais , Ciclina B1/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Fator de Iniciação 1 em Eucariotos/genética , Fator de Iniciação 1 em Eucariotos/metabolismo , Fase G1/efeitos dos fármacos , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Cinética , Leupeptinas/farmacologia , Camundongos , Camundongos Endogâmicos ICR , Inibidores de Proteassoma , Proteínas/genética , Proteínas/metabolismo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Zigoto/metabolismo , Zigoto/ultraestrutura
16.
J Reprod Dev ; 56(6): 607-15, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20710121

RESUMO

We investigated the contribution of phosphorylated RNA polymerase II (RNAP II) and dynamic epigenetic changes to the onset of minor zygotic gene activation (ZGA). Using immunofluorescence staining, we observed that the nuclear localization of RNAP II was initiated by 6 hours post insemination (hpi), whereas RNAP II phosphorylated at serine residue 5 of the carboxyl-terminal domain (CTD) was localized by 9 hpi, and then RNAP II phosphorylated at serine residue 2 of the CTD was localized in the nucleus of embryos by 12 hpi. In a transient gene expression assay using a plasmid reporter gene (pß-actin/luciferase+/SV40) injected during 6-9 hpi into the male pronucleus, the luciferase+ gene was actively transcribed and translated by 13 and 15 hpi, respectively, indicating that a transcriptionally silent state persisted for at least 4 hours after injection. We found that the methylation status in the chicken ß-actin promoter region of the plasmid reporter gene may not be associated with the transcriptionally silent state before minor ZGA. Exposure to trichostatin A did not induce premature expression of the silent reporter gene injected into 1-cell embryos containing histone deacetylase activity and did not affect the amount of luciferase produced per embryo. Acetylated histone H3 lysine 9/14 and acetylated histone H4 lysine 12 and 16 were enriched preferentially in the injected reporter gene at least until 13 hpi, which coincided with the transcriptionally active state. Taken together, these results suggest that deposition of selectively acetylated histones onto the chromatin of 1-cell embryos functions together with transcriptional elongation by RNAP II and that this sequential chromatin remodeling is involved in the molecular mechanism associated with the onset of minor ZGA in the preimplantation mouse embryo.


Assuntos
Blastocisto/metabolismo , Epigênese Genética , Histonas/metabolismo , Regiões Promotoras Genéticas , RNA Polimerase II/metabolismo , Ativação Transcricional , Zigoto/metabolismo , Acetilação/efeitos dos fármacos , Animais , Epigênese Genética/efeitos dos fármacos , Feminino , Genes Reporter/efeitos dos fármacos , Inibidores de Histona Desacetilases/farmacologia , Ácidos Hidroxâmicos/farmacologia , Cinética , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fosforilação , Regiões Promotoras Genéticas/efeitos dos fármacos , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Transporte Proteico , RNA Polimerase II/genética , Ativação Transcricional/efeitos dos fármacos , Zigoto/citologia , Zigoto/efeitos dos fármacos
17.
Biol Open ; 8(10)2019 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-31640975

RESUMO

During the maternal-to-zygotic transition (MZT), mRNAs and proteins stored in oocytes are degraded and zygotic genes are activated. We have previously shown that the ubiquitin-proteasome system (UPS)-mediated degradation of maternal proteins plays a role in the onset of zygotic transcription. However, it is still unclear which maternal proteins should be degraded for zygotic genome activation and ensuring subsequent embryonic development. In this study, we screen for these maternal factors that are degraded via the UPS. We thus identified a maternal protein PIASy (protein inhibitor of activated STATy), which is an E3 SUMO ligase. The overexpression of PIASy in fertilized embryos causes developmental arrest at the two-cell stage due to severe abnormal chromosome segregation and impaired zygotic transcription. We find that this developmental role of PIASy is related to its SUMOylation activity. Moreover, PIASy overexpression leads to increased trimethylation of histone H3 lysine 9 (H3K9me3) in two-cell nuclei and enhanced translocation of H3K9me3 methyltransferase to the pronucleus. Hence, PIASy is a maternal factor that is degraded after fertilization and may be important for the proper induction of zygotic genome activation and embryonic development.

18.
Rev Sci Instrum ; 90(7): 074707, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31370502

RESUMO

In this work, an advantage actor critic (A2C) based intelligent automatic frequency control (AFC) system was developed for X-band linear accelerator (LINAC). A2C is one type of reinforcement learning, which indicates how software agents should perform actions in an environment. In this paper, the A2C based AFC algorithm and its environment design, simulation result, and controller hardware and software processes are described. The objective of our design is to match LINAC and magnetron frequency, and it is implemented via reward shaping using comparison with the reflected power in the adjacent time. The simulation with the A2C algorithm was implemented in two modes, namely, periodic and White Gaussian Noise (WGN) waves, for analysis with temperature and random disturbance, respectively. In order to create artificial disturbance in the experiment, the magnetron shaft was shifted randomly every 0.5 s by using WGN with 18° angle of the step motor. The standard deviation of the reflected power was 5.63 kW, and the average power was 130.9 kW. To obtain maximum reward at the beginning of the A2C training, the adjacent frequency is needed. The measured average reflected power and standard deviation were 122.8 kW and 1.75 kW, respectively, after 2000 iterations. The results show that the reflected power and standard deviation of the AFC with A2C were lower compared to open-loop with artificial disturbance. The RF station of a medical X-band LINAC was used as the test bench, and the performance was confirmed by the results of an experiment conducted at Sungkyunkwan University in Korea.

19.
Nat Commun ; 10(1): 2926, 2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31249315

RESUMO

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

20.
Nat Commun ; 10(1): 2271, 2019 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118423

RESUMO

Following fertilization, cortical granules exocytose ovastacin, a metalloendopeptidase that cleaves ZP2 in the zona pellucida surrounding mouse eggs to prevent additional sperm binding. Using high- and super-resolution imaging with ovastacinmCherry as a fluorescent marker, we characterize cortical granule dynamics at single granule resolution in transgenic mouse eggs. Newly-developed imaging protocols provide an unprecedented view of vesicular dynamics near the plasma membrane in mouse eggs. We discover that cortical granule anchoring in the cortex is dependent on maternal MATER and document that myosin IIA is required for biphasic trafficking to the plasma membrane. We observe local clearance of cortical actin during exocytosis and determine that pharmacologic or genetic disruption of trafficking to the plasma membrane impairs secretion of cortical granules and results in polyspermy. Thus, the regulation of cortical granule dynamics at the cortex-plasma membrane interface is critical for exocytosis and the post-fertilization block to sperm binding that ensures monospermic fertilization.


Assuntos
Grânulos Citoplasmáticos/metabolismo , Exocitose/fisiologia , Interações Espermatozoide-Óvulo/fisiologia , Zona Pelúcida/metabolismo , Animais , Antígenos/metabolismo , Membrana Celular/metabolismo , Proteínas do Ovo/metabolismo , Feminino , Microscopia Intravital , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Masculino , Metaloproteases/química , Metaloproteases/genética , Metaloproteases/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência , Glicoproteínas da Zona Pelúcida/metabolismo , Proteína Vermelha Fluorescente
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