Short germ insects utilize both the ancestral and derived mode of Polycomb group-mediated epigenetic silencing of Hox genes

In insect species that undergo long germ segmentation, such as Drosophila, all segments are specified simultaneously at the early blastoderm stage. As embryogenesis progresses, the expression boundaries of Hox genes are established by repression of gap genes, which is subsequently replaced by Polycomb group (PcG) silencing. At present, however, it is not known whether patterning occurs this way in a more ancestral (short germ) mode of embryogenesis, where segments are added gradually during posterior elongation. In this study, two members of the PcG family, Enhancer of zeste (E(z)) and Suppressor of zeste 12 (Su(z)12), were analyzed in the short germ cricket, Gryllus bimaculatus. Results suggest that although stepwise negative regulation by gap and PcG genes is present in anterior members of the Hox cluster, it does not account for regulation of two posterior Hox genes, abdominal-A (abd-A) and Abdominal-B (Abd-B). Instead, abd-A and Abd-B are predominantly regulated by PcG genes, which is the mode present in vertebrates. These findings suggest that an intriguing transition of the PcG-mediated silencing of Hox genes may have occurred during animal evolution. The ancestral bilaterian state may have resembled the current vertebrate mode of regulation, where PcG-mediated silencing of Hox genes occurs before their expression is initiated and is responsible for the establishment of individual expression domains. Then, during insect evolution, the repression by transcription factors may have been acquired in anterior Hox genes of short germ insects, while PcG silencing was maintained in posterior Hox genes

Involvement of dachshund and Distal-less in distal pattern formation of the cricket leg during regeneration

Cricket nymphs have the remarkable ability to regenerate a functional leg following amputation, indicating that the regenerating blastemal cells contain information for leg morphology. However, the molecular mechanisms that underlie regeneration of leg patterns remain poorly understood. Here, we analyzed phenotypes of the tibia and tarsus (three tarsomeres) obtained by knockdown with regeneration-dependent RNA interference (rdRNAi) against Gryllus dachshund (Gb'dac) and Distal-less (Gb'Dll). We found that depletion of Gb'Dll mRNA results in loss of the tarsal segments, while rdRNAi against Gb'dac shortens the tibia at the two most distal tarsomeres. These results indicate that Gb'Dll expression is indispensable for formation of the tarsus, while Gb'dac expression is necessary for elongation of the tibia and formation of the most proximal tarsomere. These findings demonstrate that mutual transcriptional regulation between the two is indispensable for formation of the tarsomeres, whereas Gb'dac is involved in determination of tibial size through interaction with Gb'ds/Gb'ft

Regulation of JH biosynthesis by TGF-β signaling

While butterflies undergo a dramatic morphological transformation from larvae to adult via a pupal stage (e.g., holometamorphosis), crickets undergo a metamorphosis from nymph to adult without formation of a pupa (e.g., hemimetamorphosis). Despite these differences, both processes are regulated by common mechanisms that involve 20-hydroxyecdysone (20E) and juvenile hormone (JH). JH regulates many aspects of insect physiology, such as development, reproduction, diapauses, and metamorphosis. Consequently, strict regulation of JH levels is crucial throughout an insect’s life cycle. However, it remains unclear how JH synthesis is regulated. Here, we report that in the corpora allata (CA) of the cricket, Gryllus bimaculatus (Gb), Myoglianin (Gb’Myo), a homolog of Drosophila Myoglianin/vertebrate GDF8/11, is involved in the down-regulation of JH production by suppressing expression of a gene encoding JH acid O-methyltransferase, Gb’jhamt. In contrast, JH production is up-regulated by Decapentaplegic (Gb’Dpp) and Glass bottom boat/60A (Gb’Gbb) signaling that occurs as part of the transcriptional activation of Gb’jhamt. Gb’Myo defines the nature of each developmental transition by regulating JH titre and the interactions between JH and 20E. When Gb’myo expression is suppressed, activation of Gb’jhamt expression and secretion of 20E induces molting, thereby leading to the next instar prior to the last nymphal instar. Conversely, high Gb’myo expression induces metamorphosis during the last nymphal instar due to cessation of JH synthesis. Gb’myo also regulates final insect size. Since Myoglianin/GDF8/11 and Dpp/BMP2/4-Gbb/BMP5-8 are conserved in both invertebrates and vertebrates, the present findings provide common regulatory mechanisms regarding endocrine control of animal development

Roles of OA1 octopamine receptor and Dop1 dopamine receptor in mediating appetitive and aversive reinforcement revealed by RNAi studies

Revealing reinforcing mechanisms in associative learning is important for elucidation of brain mechanisms of behavior. In mammals, dopamine neurons are thought to mediate both appetitive and aversive reinforcement signals. Studies using transgenic fruit-flies suggested that dopamine neurons mediate both appetitive and aversive reinforcements, through the Dop1 dopamine receptor, but our studies using octopamine and dopamine receptor antagonists and using Dop1 knockout crickets suggested that octopamine neurons mediate appetitive reinforcement and dopamine neurons mediate aversive reinforcement in associative learning in crickets. To fully resolve this issue, we examined the effects of silencing of expression of genes that code the OA1 octopamine receptor and Dop1 and Dop2 dopamine receptors by RNAi in crickets. OA1-silenced crickets exhibited impairment in appetitive learning with water but not in aversive learning with sodium chloride solution, while Dop1-silenced crickets exhibited impairment in aversive learning but not in appetitive learning. Dop2-silenced crickets showed normal scores in both appetitive learning and aversive learning. The results indicate that octopamine neurons mediate appetitive reinforcement via OA1 and that dopamine neurons mediate aversive reinforcement via Dop1 in crickets, providing decisive evidence that neurotransmitters and receptors that mediate appetitive reinforcement indeed differ among different species of insects

Chalcopyrite ZnSnSb_2: A Promising Thermoelectric Material

Ternary compounds with a tetragonal chalcopyrite structure, such as CuGaTe2, are promising thermoelectric (TE) materials. It has been demonstrated in various chalcopyrite systems, including compounds with quaternary chalcopyrite-like structures, that the lattice parameter ratio, c/a, being exactly 2.00 to have a pseudo-cubic structure is key to increase the degeneracy at the valence band edge and ultimately achieve high TE performance. Considering the fact that ZnSnSb_2 with a chalcopyrite structure is reported to have c/a close to 2.00, it is expected to have multiple valence bands leading to a high p-type zT. However, there are no complete investigations on the high temperature TE properties of ZnSnSb_2 mainly because of the difficulty of obtaining a single-phase ZnSnSb_2. In the present study, pure ZnSnSb_2 samples with no impurities are synthesized successfully using a Sn flux-based method and TE properties are characterized up to 585 K. Transport properties and thermal analysis indicate that the structure of ZnSnSb_2 remains chalcopyrite with no order–disorder transition and clearly show that ZnSnSb_2 can be made to exhibit a high zT in the low-to-mid temperature range through further optimization

<Preliminary>Surface Characterization of Wood Charcoal under Different Carbonization Conditions

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Microstructural study of carbonized wood after cell wall sectioning

Wooden blocks of Japanese cedar (Cryptomeria japonica) were carbonized at 700 and 1,800 degrees C. The microstructure was analyzed by transmission electron microscopy (TEM) and mu-Raman spectroscopy of the inner planes of wood cell walls. The predominant structure was of a turbostratic nature and no heterogeneity was observed originating from the original cell walls. TEM observations of samples carbonized at 1,800 degrees C showed ordered regions in the surface layer of cell walls. This result was supported by polarized mu-Raman analysis. It may be caused by the deposition of carbon compounds volatilized from the cell walls during pulse current heating