Identical sequences but different expression patterns of Hira gene in gynogenetic and gonochoristic crucian carps

Hir/Hira (histone regulation) genes were first identified in yeast as negative regulators of histone gene expression. It has been confirmed that HIRA is a conserved family of proteins present in various animals and plants. In this paper, the cDNAs of the Hira homolog named CagHira and CaHira were isolated from gynogenetic gibel carp (gyno-carp) and gonochoristic color crucian carp (gono-carp) respectively. The full-length CagHira is 3,860 bp in length with an open reading frame (ORF) of 3,033 bp that encodes 1,011 amino acids, while the full-length CaHira is 3,748 bp in length and also has an ORF of 3,033 bp. The deduced amino acid sequences of both Hira homologs contain seven WD domains and show high identity with other HIRA family members. RT-PCR analyses revealed strong expression of Hira in the ovaries, whereas no expression was detected in the testes of either of the fishes. Hira transcription was not detected in the liver of gyno-carp, but a high level of Hira mRNA was observed in gono-carp. The temporal expression pattern showed that the Hira mRNA is consistently expressed during all embryonic development stages in gyno-carp. However, the abundance of CaHira mRNA significantly decreased (P < 0.05) shortly after fertilization and then increased again and remained stable from gastrula till hatching. The varying spatiotemporal expression patterns of Hira genes in gyno-carp and gono-carp may be associated with the differing reproductive modes used by these two closely related fishes. Our results suggest that Hira may play a role not only in the decondensation of sperm nucleus and the formation of pronucleus during fertilization, but also in gastrulation and the subsequent development of embryos.

Karyotypic diversity in polyploid gibel carp, Carassius auratus gibelio Bloch

Polyploid gibel carp, Carassius auratus gibelio, is an excellent model system for evolutionary genetics owing to its specific genetic background and reproductive modes. Comparative karyotype studies were performed in three cultured clones, one artificially manipulated group, and one mated group between two clones. Both the clones A and P had 156 chromosomes in their karyotypes, with 36 metacentric, 54 submetacentric, 36 subtelocentric, 24 acrocentric, and six small chromosomes. The karyotype of clone D contained 162 chromosomes, with 42 metacentric, 54 submetacentric, 36 subtelocentric, 24 acrocentric, and six small chromosomes. All the three clones had six small chromosomes in common. Group G, being originated from the clone D by artificial manipulation, showed supernumerary microchromosomes or chromosomal fragments, in addition to the normal chromosome complement that was identical to the clone D. The offspring from mating between clones D and A had 159 chromosomes. Comparing with the clone A, the DA offspring showed three extra metacentric chromosomes. In addition, variable RAPD fingerprint patterns and unusual SCAR marker inheritance were, respectively, detected among individuals of artificial group G and in the mated DA offspring. Both the chromosome and molecular findings suggest that genome reshuffling might have occurred by manipulation or mating of the clones.

Molecular analysis of silver crucian carp (Carassius auratus gibelio Bloch) clones by SCAR markers

Random amplified polymorphic DNA (RAPD) molecular markers specific for one, two or three clones have been identified from five gynogenetic clones of silver crucian carp (Carassius auratus gibelio Bloch) using RAPD markers developed earlier. In this study, three RAPD markers (RA1-PA, RA2-EF and RA4-D) produced by Opj-1, and two RAPD DNA fragments (RA3-PAD and RA5-D) produced by Opj-7, were selected for molecular cloning and sequencing. Sequence data indicated that there were identical 801-bp nucleotide sequences in the shared marker RA1-PA cloned respectively from clones P and A, and the shared marker RA2-EF (which was cloned from clones E and F), were also of identical 958-by nucleotide sequences. The nucleotide sequences of the shared marker RA3-PAD fragments were also similar for 1181 by among clones P, A and D. The specific fragment RA4-D was composed of 628 bp, and the fragment RA5-D from clone D contained 385 nucleotides. According to the nucleotide sequences, we designed and synthesized five pairs of sequence characterized amplified regions (SCAR) primers to identify the specific fragments in these gynogenetic clones of silver crucian carp. Only individuals from clones P and A amplified a specific band using a pair of SCI-PA primers synthesized according to the marker RA1-PA sequences, whereas no products were detected in individuals from clones D, E and F. The PCR products amplified using SC2-EF and SC3-PAD primers were as expected. Furthermore, the pair of SC4-D primers amplified specific bands only in individuals from clone D, although weak bands could be produced in all individuals of the five clones when lower annealing temperatures were used. However, an additional pair of SC5-D primers designed from the RA5-D marker sequences could amplify a DNA band in individuals from clones P, A and D, and the same weak band was produced in clone E, whereas no products were detected in individuals from clone F. Searches in GenBank revealed that the 385-bp DNA fragment from RA5-D was homologous to the 5' end of gonadotropin I beta subunit 2 gene and growth hormone gene. No homologous sequences were found for other markers in GenBank. The SCAR markers identified in this study will offer a powerful, easy, and rapid method for discrimination of different clones and for genetic analyses that examine their origins and unique reproductive modes in crucian carp. Furthermore, they will likely benefit future selective breeding programs as reliable and reproducible molecular markers. (C) 2001 Elsevier Science B.V. All rights reserved.

Genetic evidence for gonochoristic reproduction in gynogenetic silver crucian carp (Carassius auratus gibelio bloch) as revealed by RAPD assays

Sex evolution has been a debating focus in evolutionary genetics. In lower vertebrates of reptiles, amphibians, and fish, a species or a bioform reproduces either sexually or asexually but never both. A few species were found to consist of all females in fish. These all-female species can propagate by asexual reproduction modes, such as gynogenesis and hybridogenesis. However, the coexistence of sexuality and asexuality in a single species was recently noted only in a cyprinid fish silver crucian carp, Carassius auratus gibelio. This fish had been demonstrated to be capable of gynogenesis stimulated by sperm from other related species. Surprisingly, natural populations of this fish consist of a minor but significant portion (approx. 20%) of males. As different clones with specific phenotypic and genetic characteristics have been found, and RAPD markers specific to each clone have recently been identified, this fish offers many advantages for analyzing whether or not genetic recombination occurs between different clones. In this study, artificial propagation was performed in clone F and clone D. Ovulated eggs from clone F were divided into two parts and respectively inseminated with sperm from a clone D male and from a red common carp (Cyprinus carpio) male. The control clone D individuals were selected from gynogenetic offspring of clone D activated by sperm of red common carp. The phenotype and sex ratio in the experimental groups were also observed. Using RAPD molecular markers, which allow for reliable discrimination and genetic analysis of different clones, we have revealed direct molecular evidence for gonochoristic reproduction in the gynogenetic silver crucian carp and confirmed a previous hypothesis that the silver crucian carp might reproduce both gynogenetically and gonochoristically. Therefore, we conclude that the silver crucian carp possesses two reproductive modes, i.e., gynogenetic and gonochoristic reproduction. The response mechanism of two reproductive development modes may be the first discovery in vertebrates. Additionally, we discuss the evolutionary implication between gynogenetic and gonochoristic reproduction modes and the contribution of the minor proportion of males to genetic flexibility in the gynogenetic silver crucian carp.

Comparative investigation on spindle behavior and MPF activity changes during oocyte maturation between gynogenetic and amphimictic crucian carp

The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimictic colored crucian carp. MPF activity was measured by using histone I-Il as phosphorylation substrate. There were two similar oscillatory MPF kinase activity changes during oocyte maturation in two kinds of fishes with different reproductive modes, but there existed some subtle difference between them. The subtle difference was that the first peak of MPF kinase activity was kept to a longer-lasting time in the gynogenetic silver crucian carp than in the amphimictic colored crucian carp. It was suggested that the difference may be related to the spindle behavior changes, such as tripolar spindle formation and spindle rearrangement in the gynogenetic crucian carp.

Failure modes of doubly supported capacitive RF MEMS switches

For the design of radio frequency micro-electro-mechanical systems (RF MEMS) switches, the reliability issue becomes increasingly important. This paper represents some failure phenomena of doubly supported capacitive RF MEMS switches that include observable destruction failure and directly measurable parameter degradation obtained from the actuating-voltage testing and scanning electron microscope (SEM) observation. The relevant failure modes as well as their failure mechanisms are identified.

On failure modes and strength characterization of brittle disordered materials under uniaxial compression and tension

With a newly developed Material Failure Process Analysis code (MFPA(2D)), influence of hetero geneity on fracture processes and strength characterization of brittle disorder materials such as rock or concrete is numerically studied under uniaxial compression and tension conditions. It is found th at, due to the heterogeneity of the disordered material, relatively more diffused micro-fractures appear in the early stage of loading. Different from homogeneous materials such as glass, macro-crack nucleation starts well before the peak stress is reached and the crack propagation and coalescence can be traced, which can be taken as a precursory to predict the macro-fracture of the material. The presence of residual strength in the post-peak region and the resemblance in the stress-strain curves between tension and compression are significant results and are found to be dependent on the heterogeneity of the specimens. Examples showing the tentative applications of MFPA(2D) in modeling failure of composite materials and rock or civil engineering problem are also given in this paper.

Specklegram in a multiple-mode fiber and its dependence on longitudinal modes of the laser source

A specklegram in a multimode fiber (MMF) has successfully been used as a sensor for detecting external disturbance. Our experiments showed that the sensitivity in the sensor with a multiple longitudinal-mode laser as its source was much higher than that with a single longitudinal-mode laser. In addition, the near-field pattern observations indicated that the coupling between different transverse modes in the MMF is quite weak. Based on the experimental results, a theoretical model for the speckle formation is proposed, taking a bend-caused phase factor into consideration. It is shown in the theoretical analysis that the interferences between different longitudinal modes make a larger contribution to the specklegram signals. (C) 2007 Optical Society of America.

Scattering by two degenerate anisotropic modes in square-lattice dielectric photonic crystals

We systematically investigate the square-lattice dielectric photonic crystals that have been used to demonstrate flat slab imaging experimentally. A right-handed Bloch mode is found in the left-handed frequency region by using the plane wave expansion method to analyze the photonic band structure and equifrequency contours. Using the multiple scattering theory, numerical simulations demonstrate that the left-handed mode and the right-handed mode are excited simultaneously by a point source and result in two kinds of transmitted waves. Impacted by the evanescent waves, superposition of these transmitted waves brings on complicated near field distributions such as the so-called imaging and its disappearance.

Fields of apertured polychromatic laser beams with Gaussian and Hermite-Gaussian transverse modes

Starting from the Huygens-Fresnel diffraction integral, the field expressions of apertured polychromatic laser beams with Gaussian and Hermite-Gaussian transverse modes are derived. Influence of the bandwidth on the intensity distributions of the laser beams is analyzed. It is found that when the bandwidth increases, the amplitudes and numbers of the intensity spikes decrease and beam uniformity is improved in the near field and the width of transverse intensity distribution of the apertured beams decreases in the far field. Thus, the smoothing and narrowing effects can be achieved by increasing the bandwidth. Also, these effects are found in the laser beams with Hermite-Gaussian transverse modes as the bandwidth increases.(c) 2006 Elsevier Ltd. All rights reserved.

A large-area photolithography technique based on surface plasmons leakage modes

An atomic force microscope (AFM) assisted surface plasmons leakage radiation photolithography technique has been numerically demonstrated by using two-dimensional finite-difference time-domain (2D-FDTD) method. With the aid of a metallic AFM tip, particular characteristic of the Kretstchmann configuration to excite surface plasmons (SPs) is utilized to achieve large-area patterns with high spatial resolution and contrast, the photoresist could be exposed with low power laser due to the remarkable local field enhancement at the metal/dielectric interface and the resonant localized SPs modes near the tip. Good tolerance on the film thickness and incident angle has been obtained, which provides a good practicability for experiments. This photolithography technique proposed here can realize large-area, high-resolution, high-contrast, nondestructive, arbitrary-structure fabrication of nanoscale devices. (c) 2007 Elsevier B.V. All rights reserved.

Laser-induced damage threshold of the antireflection coatings on quartz and sapphire under different laser modes

The laser-induced damage threshold (LIDT) and damage morphology of antireflection (AR) coatings on quartz and sapphire are investigated. A very interesting phenomena is found in the measurement. In the case of a single pulse laser, the LIDT of the AIR coatings on quartz is higher than that of sapphire. On the contrary, for a free-pulse laser, the LIDT of AIR coatings on sapphire is higher than that of quartz. (C) 2004 Society of Photo-Optical Instrumentation Engineers.

Laser-induced damage of Ta2O5/S1O2 narrow-band interference filters under different 1064 nm Nd : YAG laser modes

The laser-induced damage (LID) behavior of narrow-band interference filters was investigated with a Nd:YAG laser at 1064 nm under single-pulse mode and free-running mode. The absorption measurement of such coatings was performed with surface thermal lensing (STL) technique. The damage morphologies under the two different laser modes were also studied in detail. It was found that all the filters exhibited a pass-band-center-dependent absorption and laser-induced damage threshold (LIDT) behavior, but the damage morphologies were diverse. The explanation was given with the analysis of the electric field distribution and the operational behavior of the irradiation laser. (c) 2005 Elsevier B.V. All rights reserved.