The sex determination cascades in insects are diversified at the top of the cascade, where different primary molecular signals are employed, while at the bottom of the cascades, particularly the doublesex genes, are highly conserved. Here, we identified the doublesex ortholog (Btau-dsx) of Bactrocera tau, a pumpkin fruit fly, and found that Btau-dsx is composed of six exons and five introns with an additional short "m" exon located in the second intron. Btau-dsx is different from its orthologs in most dipteran insects: Its pre-mRNA is sex-specifically spliced to yield three (two male and one female) instead of two transcript variants. The two deduced proteins produced by the male-specific transcripts are a functional (Btau-DSXM1) and a truncated (Btau-DSXM2) protein, while the female-specific transcript produces the functional Btau-DSXF protein. These three proteins contain all conserved domains except Btau-DSXM2 which has no OD2 domain. The female-specific transcript is detected in both fertilized and unfertilized eggs and in both somatic and germ cells of the adult females, while the male-specific transcript is detected only in fertilized eggs and in the abdominal tissues and testes of adult males. The presence of the Btau-dsxM1 transcript in fertilized eggs at the early syncytium stage suggests that in XY embryos, the Y-linked M factor gene may function quite soon after fertilization to alter the splicing pattern of Btau-dsx pre-mRNA from the female-specific to the male-specific mode. Injection of Btau-dsxF dsRNA into recently emerging females can reduce the expression of vitellogenin (Btau-Vg) and causes some defects in the ovaries, indicating that Btau-dsxF works upstream of Btau-Vg., Thanaset Thongsaiklaing, Hataichanok Passara, Mingkwan Nipitwathanaphon, Lertluk Ngersiri., and Obsahuje bibliografii
Macrocentrus cingulum is an important polyembryonic endoparasitic wasp that attacks larvae of the Asian corn borer, Ostrinia furnacalis (Guenée) and the European corn borer, O. nubilalis (Hübner). Parasitoids use antennae as the main sensory organ to recognize herbivore-induced plant volatiles as host searching cues. The antennal olfaction proteins, odorant receptors (ORs) and ionotropic receptors (IRs) are involved in olfactory signal transduction pathway as a sensory neuron response. In the present study, we constructed a cDNA library from the male and female antennae for identifying the olfaction-related genes in M. cingulum. For that, we sequenced 3160 unique gene sequences and annotated them with gene ontology (GO), cluster of orthologous groups of proteins (COG), and KEGG ontology (KO). Through the homology search, we identified 9 odorant receptors (ORs), 3 ionotropic receptors (IRs) and 1 odorant binding protein (OBP) genes from the cDNA library sequences. Additionally, the expression patterns of these ORs and IRs in different tissues (antennae, heads, thoraxes, abdomens, and legs) were demonstrated by RT-PCR. The qualitative gene expression analyses showed that most of the OR genes were more highly expressed in female than male antennae; whereas IRs, unlike ORs, were more expressed in various male than females tissues. We are the first to report ORs and IRs in M. cingulum, which should help in deciphering the molecular basis of olfaction system in this wasp., Tofael Ahmed, Tian-Tao Zhang, Zhen-Ying Wang, Kang-Lai He, Shu-Xiong Bai., and Obsahuje bibliografii
Novel microsatellite markers for the spruce bark beetle (Ips typographus L.) are identified and characterized using next generation sequencing technology. 18 polymorphic loci were obtained by screening 10,684 reads and tested on 35 bark beetle samples from different locations in Bavaria, Germany. Allelic richness ranged from two to 38 alleles, observed heterozygosity from 0.03 to 0.66 and expected heterozygosity from 0.08 to 0.97. Four loci showed significant deviation from the Hardy-Weinberg equilibrium and no linkage between loci was detected. The 18 loci, along with another six loci previously described, provide effective analytical tools for analyzing the fine-scale genetic structure of bark beetle populations. The result of this study demonstrates that next generation sequencing technology is a valuable method for isolating microsatellites of a coleopteran species. and Bernhard C. STOECKLE, Ralph KUEHN.
Novel microsatellite markers for the solitary bee, Anthophora plumipes, were identified and characterised using 454 GS-FLX Titanium pyrosequencing technology. Thirty seven loci were tested using fluorescently labelled primers on a sample of 20 females from Prague. The number of alleles ranged from 1 to 10 (with a mean of 4 alleles per locus), resulting in an observed heterozygosity ranging from 0.05 to 0.9 and an expected heterozygosity from 0.097 to 0.887. None of the loci showed a significant deviation from the Hardy-Weinberg equilibrium and only two loci showed the significant presence of null alleles. No linkage between loci was detected. We further provide information on a single multiplex PCR consisting of 11 of the most polymorphic loci. This multiplex approach provides an effective analytical tool for analysing genetic structure and carrying out parental analyses on Anthophora populations. Most of the 37 loci tested also showed robust amplification in five other Anthophora species (A. aestivalis, A. crinipes, A. plagiata, A. pubescens and A. quadrimaculata). The result of this study demonstrates that next generation sequencing technology is a valuable method for isolating quality microsatellites in non-model species of solitary bees. and Obsahuje seznam literatury
Tapinoma melanocephalum is a worldwide distributed, highly invasive ant species. It lives in close association with human societies and its distribution is human-mediated in large measure. The geographical origin of this ant species is unknown, but its introduction in areas previously devoided of its presence can represent a threat to the native biota, act as an agricultural pest or as a pathogen vector. To investigate the genetic structure and phylogeography of this species we identified 12 new polymorphic microsatellite markers, and in addition, we tested and selected 12 ant-universal microsatellites polymorphic in T. melanocephalum. We genotyped 30 individuals from several islands of Micronesia and Papua-New Guinea. All 24 loci exhibited strong homozygosity excess (45-100%, mean = 86%), while the number of alleles per locus reached usual values (2-18, mean = 6.5), resulting in levels of expected heterozygosity much higher than observed. Based on several robust tests, we were able to exclude artefacts such as null alleles and allelic dropout as a possible cause of the observed pattern. Homozygosity excess might be a consequence of founder effect, bottleneck and/or inbreeding. As our sample population was composed of individuals from several distinct localities, the Wahlund effect might have contributed to the increased homozygosity as well. Despite the provisionally observed deviation from the Hardy-Weinberg equilibrium, the newly developed microsatellites will provide an effective tool for future genetic investigations of population structure as well as for the phylogeographic study of T. melanocephalum., Jan Zima Jr., Ophélie Lebrasseur, Michaela Borovanská, Milan Janda., and Obsahuje bibliografii
Our study was aimed to characterize the phenotype and functional endpoints of local microwave hyperthermia (LHT, 42 °C) on tumor infiltrating and spleen leukocytes. The effectiveness of LHT applied into the tumor of B16F10 melanoma-bearing C57/BL6 mice was compared with anesthetized and non-treated animals. Subpopulations of leukocytes were analyzed using the flow cytometry, and the cytotoxic activity of splenocytes against syngeneic B16F10 melanoma and NK-sensitive YAC-1 tumor cell lines was evaluated in 51 Cr-release assay. Similarly, the in vitro modification of the heat treatment was performed using healthy and melanoma-bearing splenocytes. We found a 40 % increase of activated monocytes (CD11b+CD69+) infiltration into the tumor microenvironment. In the spleen of experimental animals, the numbers of cytotoxic T lymphocytes (CTLs-CD3+CD8+) and NK cell (CD49b+NK1.1+) raised by 22 % and 14 %, respectively, while the NK1.1+ monocytes decreases by 37 %. This was accompanied by an enhancement of cytotoxic effector function against B16F10 and YAC-1 targets in both in vivo and in vitro conditions. These results demonstrate that LHT induces better killing of syngeneic melanoma targets. Furthermore, LHT evokes the homing of activated monocytes into the tumor microenvironment and increases the counts of NK cells and CTL in the spleen., J. Kubeš, J. Svoboda, J. Rosina, M. Starec, A. Fišerová., and Obsahuje bibliografii a bibliografické údaje
The effects of different abiotic agents that may modulate the activity of an insect’s immune system are reviewed. These agents include insecticides, chitin synthesis inhibitors, juvenile hormone analogues, inert particles, antibiotics, heavy metals, radiation and miscellaneous substances. The significance of studying immunomodulation in insects treated with abiotic agents in relation to both insect control and insect-borne parasitic diseases and the link between immunomodulation in insects post-treatment with both abiotic and biotic agents are discussed., El-Sayed H. Shaurub., and Obsahuje seznam literatury
Among animal constructions, the orb webs of spiders are perfect geometrical architectural models. Webs are constructed by means of a succession of behaviours, which are often considered as instinctive and stereotyped. Anomalies in web structure are considered to be a consequence of variability in behaviour. In order to assess the variability in web structure, the spider Zygiella x-notata was allowed to build webs in the laboratory under standardized conditions. Webs (n = 127) were examined for anomalies, which were defined as modifications in the geometrical structure. Ten anomalies in the structure of webs produced by females, aged from 16 to 21 days after their adult moult, are described. The frequency of anomalies per web varies from 5 to 98. A very low percentage of the spiral elements of a web (less than 5%) were affected. Anomalies in the radii were less numerous (mean = 3 ± 3) than those in the sticky spiral (mean = 21 ± 11). Anomalies in the sticky spiral were different and more frequent in the peripheral than central parts of the webs. They were also relatively more numerous in the upper than in the lower part of the webs. Total number of anomalies was positively correlated with particular web characteristics (capture area, length of capture thread, width of the lower part of the web and mean distance between two spiral turns in the lower part) but not to the physical characteristics of the spiders (mass and size). Our results indicate that the orb webs of spiders, which are considered to be perfect geometrical structures, include anomalies mainly in the structure of the sticky spiral. These anomalies were the result of variability in behaviour during web building. In conclusion, their identification and description enable one to use them in spider webs as an index of behavioural variability., Alain Pasquet ... [et al.]., and Obsahuje seznam literatury
Animals, including human beings, tend to respond more strongly to stimuli that are associated with the highest relative rewards. This applies not only to food rewards but also to reproductive success. In the present review article this issue is discussed for insects in connection with intersexual communication and flower-visiting behaviour. Implications of the preference for supernormal visual releasing stimuli are examined from a sensory and evolutionary perspective, including a consideration of the choice of potential mates and recognition of the most rewarding flowers., Karl Kral., and Obsahuje bibliografii
Chronic hypoxia results in hypoxic pulmonary hypertension characterized by fibrotization and muscularization of the walls of peripheral pulmonary arteries. This vessel remodeling is accompanied by an increase in the amount of lung mast cells (LMC) and the presence of small collagen cleavage products in the vessel walls. We hypothesize that hypoxia activates LMC, which release matrix metalloproteinases (MMPs) cleaving collagen and starting increased turnover of connective tissue proteins. This study was designed to determine whether in vitro hypoxia stimulates production of MMPs in rat LMC and increases their collagenolytic activity. The LMC were separated on the Percoll gradient and then were divided into two groups and cultivated for 24 h in 21 % O2 + 5 % CO2 or in 10 % O2 + 5 % CO2. Presence of the rat interstitial tissue collagenase (MMP-13) in LMC was visualized by immunohistological staining and confirmed by Western blot analysis. Total MMPs activity and tryptase activity were measured in both cultivation media and cellular extracts. Exposure to hypoxia in vitro increased the amount of cells positively labeled by anti-MMP-13 antibody as well as activities of all measured enzymes. The results therefore support the concept that LMC are an important source of increased collagenolytic activity in chronic hypoxia., H. Maxová, J. Novotná, L. Vajner, H. Tomášová, R. Vytášek, M. Vízek, L. Bačáková, V. Valoušková, T. Eliášová, J. Herget., and Obsahuje bibliografii a bibliografické odkazy