Regulation of zygotic gene activation in the preimplantation mouse embryo: Global activation and repression of gene expression

Superimposed on the activation of the embryonic genome in the preimplantation mouse embryo is the formation of a transcriptionally repressive state during the two-cell stage. This repression appears mediated at the level of chromatin structure, because it is reversed by inducing histone hyperacetylation or inhibiting the second round of DNA replication. We report that of more than 200 amplicons analyzed by mRNA differential display, about 45% of them are repressed between the two-cell and four-cell stages. This repression is scored as either a decrease in amplicon expression that occurs between the two-cell and four-cell stages or on the ability of either trichostatin A tan inhibitor of histone deacetylases) or aphidicolin tan inhibitor of replicative DNA polymerases) to increase the level of amplicon expression. Results of this study also indicate that about 16% of the amplicons analyzed likely are novel genes whose sequence doesn't correspond to sequences in the current databases, whereas about 20% of the sequences expressed during this transition likely are repetitive sequences. Lastly, inducing histone hyperacetylation in the two-cell embryos inhibits cleavage to the four-cell stage. These results suggest that genome activation is global and relatively promiscuous and that a function of the transcriptionally repressive state is to dictate the appropriate profile of gene expression that is compatible with further development.

Identification of differential transcript profiles between mutual crossbred embryos of zebrafish (Danio rerio) and Chinese rare minnow (Gobiocypris rarus) by cDNA-AFLP

The crosstalk between naive nucleus and maternal factors deposited in egg cytoplasm before zygotic genome activation is crucial for early development. In this study, we utilized two laboratory fishes, zebrafish (Danio rerio) and Chinese rare minnow and Chinese rare minnow (Gobiocypris rarus), to obtain mutual crossbred embroys and examine the interaction between nucleus and egg cytoplasm from different species. Although these two types of crossbred embryos originated from common nuclei, various developmental capacities were gained due to different origins of the egg cytoplasm. Using cDNA amplified fragment length polymorphism (cDNA-AFLP), We Compared transcript profiles between the mutual crossbred embryos at two developmental stages (50%- and 90%-epiholy). Three thousand cDNA fragments were generated in four cDNA pools with 64 primer combinations. All differently displayed transcript-derived fragments (TDFs) were screened by (lot blot hybridization, and the selected sequences were further analyzed by semi-quantitative RT-PCR and quantitative real-time RT-PCR. Compared with ZR embryos, 12 genes were up-regulated and 12 were down-regulated in RZ embryos. The gene fragments were sequenced and subjected to BLASTN analysis. The sequences encoded various proteins which functioned at various levels of proliferation, growth, and development. One gene (ZR6), dramatically down-regulated in RZ embryos, was chosen for loss-of-function study; the knockdown of ZR6 gave rise to the phenotype resembling that of RZ embryos. (c) 2008 Elsevier Inc. All rights reserved.

Constitutive expression of thymidylate synthase from LCDV-C induces a transformed phenotype in fish cells

Thymidylate synthase (TS), an essential enzyme in DNA synthesis and repair, plays a key role in the events of cell cycle regulation and tumor formation. Here, an investigation was presented about subcellular location and biological function of viral TS from lymphocystis disease virus from China (LCDV-C) in fish cells. Fluorescence microscopy revealed that LCDV-C TS was predominantly localized in the cytoplasm in fish cells. Cell cycle analysis demonstrated that LCDV-C TS promoted cell cycle progression into S and G2/M phase in the constitutive expressed cells. As a result, the cells have a faster growth rate compared with the control cells as revealed by cell growth curves. For foci assay, the TS-expressed cells gave rise to foci 4-5 weeks after incubation. Microscopic examination of the TS-induced foci revealed multilayered growth and crisscross morphology characteristic of transformed cells. Moreover, LCDV-C TS predisposed the transfected cells to acquire an anchorage-independent phenotype and could grow in 0.3% soft agar. So the data reveal LCDV-C TS is sufficient to induce a transformed phenotype in fish cells in vitro and exhibits its potential ability in cell transformation. To our knowledge, it is the first report on viral TS sequences associated with transforming activity. (C) 2007 Elsevier Inc. All rights reserved.

Free radical scavenging and antioxidant enzymes activation of polysaccharide extract from Nostoc sphaeroides

Nostoc sphaeroides Kuetzing has been used as a traditional medicine in China to treat a variety of ailments. This research identified the antioxidant activities of polysaccharide extract from Nostoc sphaeroides. The extract, which contains 46.2% carbohydrates, exhibited an effective scavenging capability on superoxide radical, hydroxyl radicals in non site-specific as well as site-specific assays, and also performed lipid peroxidation inhibition in a dose-dependent manner. Polysaccharide extract had no 1,1-diphenyl-2-picrylhydrazyl radical scavenging potential at all test concentrations. Activities of superoxide dismutase, catalase, and glutathione peroxidase in human embryo kidney 293 cells were increased effectively when Nostoc sphaeroides extract was applied. These results suggested that the use of N. sphaeroides in treating ailments may be based on the antioxidant capacities of polysaccharide composition.

Molecular identification and expression analysis of natural resistance associated macrophage protein (Nramp) cDNA from Japanese flounder (Paralichthys olivaceus)

Natural resistance associated macrophage protein (Nramp) controls partially innate resistance to intracellular parasites. Its function is to enhance the ability of macrophages to kill pathogens. However, little is known about the structure and function of Nramp in lower vertebrates such as teleosts. We have recently isolated a cDNA encoding Nramp from Japanese flounder (Paratichthys olivaceus). The full-length cDNA of the Nramp is 3066 bp in length, including 224 bp 5' terminal UTR, 1662 bp encoding region and 1180 bp 3' terminal UTR. The 1662-nt open reading frame was found to code for a protein with 554 amino acid residues. Comparison of amino acid sequence indicated that Japanese flounder Nramp consists of 12 transmembrane (TM) domains. A consensus transport motif (CTM) containing 20 residues was observed between transmembrane domains 8 and 9. The deduced amino acid sequence of Japanese flounder had 77.30%, 82.71%, 82.67%, 79.64%, 80.72%, 90.97%, 91.16%, 60.14%, 71.48%, 61.69%, 72.37% identity with that of rainbow trout Nramp alpha and beta, channel catfish Nramp, fathead minnow Nramp, common carp Nramp, striped sea bass Nramp, red sea bream Nramp, mouse Nramp 1 and 2, human Nramp 1 and 2, respectively. RT-PCR indicated that Nramp transcripts were highly abundant in spleen, head kidney, abundant in intestine, liver and gill, and less abundant in heart. The level of Nramp mRNA in embryos gradually increases during embryogenesis from 4 h (8 cell stage) to 80 h (hatched stage) after fertilization. (c) 2005 Elsevier Ltd. All rights reserved.

Annealing and activation of silicon implanted in semi-insulating InP substrates

We have investigated the annealing and activation of silicon implanted in both as-grown Fe-doped semi-insulating (SI) InP substrate and undoped SI InP substrate obtained by annealing high purity conductive InP wafer (wafer-annealed). Si implantations were performed at an energy of 500 keV and a dose of 1 X 10(15) cm(-2). Following the implantations, rapid thermal annealing (RTA) cycles were carried out for 30 s at different temperatures. The results of Raman measurements show that for 700degreesC/30s RTA, the two Si-implanted SI InP substrates have acquired a high degree of lattice recovery and electrical activation. However, further Hall measurements indicate that the carrier concentration of the wafer-annealed SI InP substrate is about three times higher than that of the as-grown Fe-doped SI InP substrate. The difference can be ascribed to the low Fe concentration of the wafer-annealed SI InP substrate.These experimental data imply that the use of the wafer-annealed SI InP substrate can be conducive to the improvement of InP-based device performances. (C) 2003 Elsevier Ltd. All rights reserved.

Solid-phase crystallization and dopant activation of amorphous silicon films by pulsed rapid thermal annealing

An improved pulsed rapid thermal annealing method has been used to crystallize amorphous silicon films prepared by PECVD. The solid-phase crystallization and dopant activation process can be completed with time-temperature budgets such as 10 cycles of 60-s 550 degrees C thermal bias/l-s 850 degrees C thermal pulse. A mean grain size more than 1000 Angstrom and a Hall mobility of 24.9 cm(2)/V s are obtained in the crystallized films. The results indicate that this annealing method possesses the potential for fabricating large-area and good-quality polycrystalline silicon films on low-cost glass substrate. (C) 1998 Elsevier Science B.V. All rights reserved.

Thermal activation and thermal transfer of localized excitons in InAs self-organized quantum dots

We have investigated the temperature dependence of photoluminescence (PL) properties of a number of InAs/GaAs heterostructures with InAs layer thickness ranging from 0.5 monolayer (ML) to 3 ML. The temperature dependence of the InAs exciton energy and linewidth was found to display a significant difference when the InAs layer thickness is smaller or larger than the critical thickness around 1.7 ML, indicating spontaneous formation of quantum dots (QDs). A model, involving exciton recombination and thermal activation and transfer, is proposed to explain the experimental data. In the PL thermal quenching study, the measured thermal activation energies of different samples demonstrate that the InAs wetting layer may act as a barrier for thermionic emission of carriers in high quality InAs multilayers, while in InAs monolayers and submonolayers the carriers are required to overcome the GaAs barrier to thermally escape from the localized states. (C) 1998 Academic Press Limited.

RECRYSTALLIZATION, IMPURITY MIGRATION AND OPTICAL ACTIVATION OF YB-IMPLANTED SILICON INDUCED BY RAPID THERMAL ANNEALING

The rapid thermal annealing temperature dependence of the recrystallization, Yb migration and its optical activation were studied for Yb-implanted silicon. For the annealing regime 800-1000-degrees-C, the Yb segregates both at the crystal/amorphous interface and at the surface, which is different from the usual segregation of Er at the crystal/amorphous interface, and the efficiency of optical activation also increases with annealing temperature. However, the amorphous layer regrows completely and no photoluminescence is observed after the annealing at 1200-degrees-C.

MBE of GaN with DC-plasma source for nitrogen activation

In this paper we report on the first results of epitaxial growth of GaN layers on GaAs (100) substrates using a modified MBE system, equipped with a DC-plasma source for nitrogen activation in configuration of reverse magnetron at ultra-low pressures.

Carrier relaxation and thermal activation of localized excitons in self-organized InAs multilayers grown on GaAs substrates

We have investigated the temperature dependence of photoluminescence (PL) properties of a number of self-organized InAs/GaAs heterostructures with InAs layer thickness ranging from 0.5 to 3 ML. The temperature dependence of InAs exciton emission and linewidth was found to display a significant difference when the InAs layer thickness is smaller or larger than the critical thickness around 1.7 ML. The fast redshift of PL energy and an anomalous decrease of linewidth with increasing temperature were observed and attributed to the efficient relaxation process of carriers in multilayer samples, resulting from the spread and penetration of the carrier wave functions in coupled InAs quantum dots. The measured thermal activation energies of different samples demonstrated that the InAs wetting layer may act as a barrier for the thermionic emission of carriers in high-quality InAs multilayers, while in InAs monolayers and submonolayers the carriers are required to overcome the GaAs barrier to escape thermally from the localized states.

Photoluminescence and activation on SI-GaAs by Si+ implantation and following rapid thermal annealing

Photoluminescence (PL) and electrical characteristics of SI-GaAs, Si+-implanted and following rapid thermal annealing (RTA), were investigated, The PL spectra of Si-GA-C-As, Ga-i-Si-As, and V-As-Si-As were obtained. This paper concentrates on the PL peak at 1.36 eV which was proven as an emission of the Si-Ga-V-Ga combination by Si+ + P+ dual implantation. The results indicate that the peak at 1.36 eV appears when the ratio of As:Ga increased during the processing. Also high activation was obtained for the sample under the same conditions. More discussion on the mechanism of Si+ implanted SI-GaAs has been made based on the Morrow model [J. Appl. Phys, 64 (1988) 1889].

Catalytic Activation of Mg-Doped GaN by Hydrogen Desorption Using Different Metal Thin Layers

The annealing of Mg-doped GaN with Pt and Mo layers has been found to effectively improve the hole concentration of such material by more than 2 times as high as those in the same material without metal. Compared with the Ni and Mo catalysts, Pt showed good activation effect for hydrogen desorption and ohmic contact to the Ni/Au electrode. Despite the weak hydrogen desorption, Mo did not diffuse into the GaNepilayer in the annealing process, thus suppressing the carrier compensation phenomenon with respect to Ni and Pt depositions, which resulted in the high activation of Mg acceptors. For the GaN activated with the Ni, Pt, and Mo layers, the blue emission became dominant, followed by a clear peak redshift and the degradation of photoluminescence signal when compared with that of GaN without metal.