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Mechanics has an important role during morphogenesis, both in the generation of forces driving cell shape changes and in determining the effective material properties of cells and tissues. Drosophila dorsal closure has emerged as a reference model system for investigating the interplay between tissue mechanics and cellular activity. During dorsal closure, the amnioserosa generates one of the major forces that drive closure through the apical contraction of its constituent cells. We combined quantitation of live data, genetic and mechanical perturbation and cell biology, to investigate how mechanical properties and contraction rate emerge from cytoskeletal activity. We found that a decrease in Myosin phosphorylation induces a fluidization of amnioserosa cells which become more compliant. Conversely, an increase in Myosin phosphorylation and an increase in actin linear polymerization induce a solidification of cells. Contrary to expectation, these two perturbations have an opposite effect on the strain rate of cells during DC. While an increase in actin polymerization increases the contraction rate of amnioserosa cells, an increase in Myosin phosphorylation gives rise to cells that contract very slowly. The quantification of how the perturbation induced by laser ablation decays throughout the tissue revealed that the tissue in these two mutant backgrounds reacts very differently. We suggest that the differences in the strain rate of cells in situations where Myosin activity or actin polymerization is increased arise from changes in how the contractile forces are transmitted and coordinated across the tissue through ECadherin-mediated adhesion. Altogether, our results show that there is an optimal level of Myosin activity to generate efficient contraction and suggest that the architecture of the actin cytoskeleton and the dynamics of adhesion complexes are important parameters for the emergence of coordinated activity throughout the tissue.
The mitochondrial kinase PINK1 and the ubiquitin ligase Parkin are participating in quality control after CCCP- or ROSinduced mitochondrial damage, and their dysfunction is associated with the development and progression of Parkinson’s disease. Furthermore, PINK1 expression is also induced by starvation indicating an additional role for PINK1 in stress response. Therefore, the effects of PINK1 deficiency on the autophago-lysosomal pathway during stress were investigated. Under trophic deprivation SH-SY5Y cells with stable PINK1 knockdown showed downregulation of key autophagic genes, including Beclin, LC3 and LAMP-2. In good agreement, protein levels of LC3-II and LAMP-2 but not of LAMP-1 were reduced in different cell model systems with PINK1 knockdown or knockout after addition of different stressors. This downregulation of autophagic factors caused increased apoptosis, which could be rescued by overexpression of LC3 or PINK1. Taken together, the PINK1-mediated reduction of autophagic key factors during stress resulted in increased cell death, thus defining an additional pathway that could contribute to the progression of Parkinson’s disease in patients with PINK1 mutations.
It was long assumed that translation initiation in prokaryotes generally occurs via the so-called Shine Dalgarno (SD) mechanism. Recently, it became clear that translation initiation in prokaryotes is more heterogeneous. In the haloarchaeon Haloferax volcanii, the majority of transcripts is leaderless and most transcripts with a 5′-UTR lack a SD motif. Nevertheless, a bioinformatic analysis predicted that 20–30% of all genes are preceded by a SD motif in haloarchaea. To analyze the importance of the SD mechanism for translation initiation in haloarchaea experimentally the monocistronic sod gene was chosen, which contains a 5′-UTR with an extensive SD motif of seven nucleotides and a length of 19 nt, the average length of 5′UTRs in this organism. A translational fusion of part of the sod gene with the dhfr reporter gene was constructed. A mutant series was generated that matched the SD motif from zero to eight positions, respectively. Surprisingly, there was no correlation between the base pairing ability between transcripts and 16S rRNA and translational efficiency in vivo under several different growth conditions. Furthermore, complete replacement of the SD motif by three unrelated sequences did not reduce translational efficiency. The results indicate that H. volcanii does not make use of the SD mechanism for translation initiation in 5′-UTRs. A genome analysis revealed that while the number of SD motifs in 5′-UTRs is rare, their fraction within open reading frames is high. Possible biological functions for intragenic SD motifs are discussed, including re-initiation of translation at distal genes in operons.
Haloferax volcanii uses extracellular DNA as a source for carbon, nitrogen, and phosphorous. However, it can also grow to a limited extend in the absence of added phosphorous, indicating that it contains an intracellular phosphate storage molecule. As Hfx. volcanii is polyploid, it was investigated whether DNA might be used as storage polymer, in addition to its role as genetic material. It could be verified that during phosphate starvation cells multiply by distributing as well as by degrading their chromosomes. In contrast, the number of ribosomes stayed constant, revealing that ribosomes are distributed to descendant cells, but not degraded. These results suggest that the phosphate of phosphate-containing biomolecules (other than DNA and RNA) originates from that stored in DNA, not in rRNA. Adding phosphate to chromosome depleted cells rapidly restores polyploidy. Quantification of desiccation survival of cells with different ploidy levels showed that under phosphate starvation Hfx. volcanii diminishes genetic advantages of polyploidy in favor of cell multiplication. The consequences of the usage of genomic DNA as phosphate storage polymer are discussed as well as the hypothesis that DNA might have initially evolved in evolution as a storage polymer, and the various genetic benefits evolved later.
Poster presentation at 1st International Workshop on Odor Spaces.
Mice are exceptional in their ability to capture their chemical environment, mapping the olfactory world into a basic sensory representation with over one thousand different types of chemical sensors, that is, olfactory sensory neurons (OSNs). OSNs of each type converge in the olfactory bulb onto exclusive distinct physiological areas called glomeruli. The glomeruli constitute the first relay station of olfactory stimulus representation in the mouse brain. Thus, the stimulus induced glomerular input pattern spatially embodies an important part of the sensory representation in the olfactory bulb. Still, topographic organization principles (chemotopy, tunotopy) are under debate. One reason might be that investigation are, due to experimental limitations, only performed on stimuli sets in the size of one hundred odors. But this represents only a tiny snapshot of the vast amount of molecules in the olfactory world and topographic relationships might be disguised in the incomplete representation of molecular receptive ranges (MRR). Therefore we investigated the problem with the MOR18-2 glomerulus as point of reference: First we determined it's MRR. Then, based on a measurement set covering this MRR, we elucidated the topographic embedding. It shows that MOR18-2 is embedded in a hierarchy of patchy tunotopic domains.
A third Supplement to the 1992 Catalog of the Scolytidae and Platypodidae (Coleoptera) of the World (Wood and Bright 1992) is presented. This Supplement updates the taxonomy, distribution, and biology pertaining to these families of Coleoptera from 2000 to the end of 2010. A few 2011 taxonomic references are included in order to make the nomenclature as current as possible. The format of this Supplement differs slightly from the format of the original 1992 Catalog and the two previous Supplements. Only references relating to general biology, systematics and distribution were selected to be included. Tribal arrangement follows the scheme established in the 1992 World Catalog, with additions. Each genus is listed alphabetically under the current tribe and the species are listed alphabetically under the current genus. Additional information on figures, distribution, hosts, and references relating to the above are given. The bibliography used with this Supplement lists 580 references, most published from 2000 to the end of 2010. New synonymy proposed: Phloeotribus carinatus Burgos and Equihua, 2003 (= Phloeotribus ebeneus Wood, 2007). New combinations proposed: Phloeosinus kinabaluensis Bright to Hyledius; Phloeosinus phyllocladus Bright to Hyledius.
An argument is made for the retention of the family status of the Scolytidae and Platypodidae as members of the superfamily Curculionoidea. A new subfamily and tribal arrangement is proposed: SCOLYTIDAE comprising 13 subfamilies, Hylesininae, with 12 tribes, Hyorrhynchinae, with one tribe; Scolytinae, with one tribe; Hexacolinae, with one tribe; Cylindrobrotinae, with one tribe; Scolytoplatypodinae, with one tribe; Cactopininae, with one tribe; Carphodicticinae, with one tribe; Micracidinae, with one tribe; Crypturginae, with one tribe; Ipinae, with six tribes; Cryphalinae, with one tribe and Corthylinae, with two tribes and PLATYPODIDAE comprising three subfamilies, Coptonotinae, with three tribes; Tesserocerinae, with two tribes and Platypodinae, with one tribe.
The use of reporter gene fusions to assess cellular processes such as protein targeting and regulation of transcription or translation is established technology in archaeal, bacterial, and eukaryal genetics. Fluorescent proteins or enzymes resulting in chromogenic substrate turnover, like β -galactosidase, have been particularly useful for microscopic and screening purposes. However, application of such methodology is of limited use for strictly anaerobic organisms due to the requirement of molecular oxygen for chromophore formation or color development. We have developed β -lactamase from Escherichia coli (encoded by bla) in conjunction with the chromogenic substrate nitrocefin into a reporter system usable under anaerobic conditions for the methanogenic archaeon Methanosarcina acetivorans. By using a signal peptide of a putative flagellin from M. acetivorans and different catabolic promoters, we could demonstrate growth substrate-dependent secretion of β -lactamase, facilitating its use in colony screening on agar plates. Furthermore, a series of fusions comprised of a constitutive promoter and sequences encoding variants of the synthetic tetracycline-responsive riboswitch (tc-RS) was created to characterize its influence on translation initiation in M. acetivorans. One tc-RS variant resulted in more than 11-fold tetracycline-dependent regulation of bla expression, which is in the range of regulation by naturally occurring riboswitches. Thus, tc-RS fusions represent the first solely cis-active, that is, factor-independent system for controlled gene expression in Archaea.
Background: Endometriosis is characterized by the presence of functional endometrial tissue outside of the uterine cavity. It affects 1 in 10 women of reproductive age. This chronic condition commonly leads to consequences such as pelvic pain, dysmenorrhea, infertility and an elevated risk of epithelial ovarian cancer. Despite the prevalence of endometriosis and its impact on women's lives, there are relatively few in vitro and in vivo models available for studying the complex disease biology, pathophysiology, and for use in the preclinical development of novel therapies. The goal of this study was to develop a novel three-dimensional (3D) cell culture model of ovarian endometriosis and to test whether it is more reflective of endometriosis biology than traditional two dimensional (2D) monolayer cultures.
Methods: A novel ovarian endometriosis epithelial cell line (EEC16) was isolated from a 34-year old female with severe endometriosis. After characterization of cells using in vitro assays, western blotting and RNA-sequencing, this cell line and a second, already well characterized endometriosis cell line, EEC12Z, were established as in vitro 3D spheroid models. We compared biological features of 3D spheroids to 2D cultures and human endometriosis lesions using immunohistochemistry and real-time semi-quantitative PCR.
Results: In comparison to normal ovarian epithelial cells, EEC16 displayed features of neoplastic transformation in in vitro assays. When cultured in 3D, EEC16 and EEC12Z showed differential expression of endometriosis-associated genes compared to 2D monolayer cultures, and more closely mimicked the molecular and histological features of human endometriosis lesions.
Conclusions: To our knowledge, this represents the first report of an in vitro spheroid model of endometriosis. 3D endometriosis models represent valuable experimental tools for studying EEC biology and the development of novel therapeutic approaches.
The haloarchaeon Haloferax volcanii was shown to contain 145 intergenic and 45 antisense sRNAs. In a comprehensive approach to unravel various biological roles of haloarchaeal sRNAs in vivo, 27 sRNA genes were selected and deletion mutants were generated. The phenotypes of these mutants were compared to that of the parent strain under ten different conditions, i.e. growth on four different carbon sources, growth at three different salt concentrations, and application of four different stress conditions. In addition, cell morphologies in exponential and stationary phase were observed. Furthermore, swarming of 17 mutants was analyzed. 24 of the 27 mutants exhibited a difference from the parent strain under at least one condition, revealing that haloarchaeal sRNAs are involved in metabolic regulation, growth under extreme conditions, regulation of morphology and behavior, and stress adaptation. Notably, 7 deletion mutants showed a gain of function phenotype, which has not yet been described for any other prokaryotic sRNA gene deletion mutant. Comparison of the transcriptomes of one sRNA gene deletion mutant and the parent strain led to the identification of differentially expressed genes. Genes for flagellins and chemotaxis were up-regulated in the mutant, in accordance with its gain of function swarming phenotype. While the deletion mutant analysis underscored that haloarchaeal sRNAs are involved in many biological functions, the degree of conservation is extremely low. Only 3 of the 27 genes are conserved in more than 10 haloarchaeal species. 22 of the 27 genes are confined to H. volcanii, indicating a fast evolution of haloarchaeal sRNA genes.
Genome-wide association studies are widely used to correlate phenotypic traits with genetic variants. These studies usually compare the genetic variation between two groups to single out certain Single Nucleotide Polymorphisms (SNPs) that are linked to a phenotypic variation in one of the groups. However, it is necessary to have a large enough sample size to find statistically significant correlations. Direct-To-Consumer (DTC) genetic testing can supply additional data: DTC-companies offer the analysis of a large amount of SNPs for an individual at low cost without the need to consult a physician or geneticist. Over 100,000 people have already been genotyped through Direct-To-Consumer genetic testing companies. However, this data is not public for a variety of reasons and thus cannot be used in research. It seems reasonable to create a central open data repository for such data. Here we present the web platform openSNP, an open database which allows participants of Direct-To-Consumer genetic testing to publish their genetic data at no cost along with phenotypic information. Through this crowdsourced effort of collecting genetic and phenotypic information, openSNP has become a resource for a wide area of studies, including Genome-Wide Association Studies. openSNP is hosted at http://www.opensnp.org, and the code is released under MIT-license at http://github.com/gedankenstuecke/snpr.
Three neonicotinoids, imidacloprid, clothianidin and thiacloprid, agonists of the nicotinic acetylcholine receptor in the central brain of insects, were applied at non-lethal doses in order to test their effects on honeybee navigation. A catch-and-release experimental design was applied in which feeder trained bees were caught when arriving at the feeder, treated with one of the neonicotinoids, and released 1.5 hours later at a remote site. The flight paths of individual bees were tracked with harmonic radar. The initial flight phase controlled by the recently acquired navigation memory (vector memory) was less compromised than the second phase that leads the animal back to the hive (homing flight). The rate of successful return was significantly lower in treated bees, the probability of a correct turn at a salient landscape structure was reduced, and less directed flights during homing flights were performed. Since the homing phase in catch-and-release experiments documents the ability of a foraging honeybee to activate a remote memory acquired during its exploratory orientation flights, we conclude that non-lethal doses of the three neonicotinoids tested either block the retrieval of exploratory navigation memory or alter this form of navigation memory. These findings are discussed in the context of the application of neonicotinoids in plant protection.
A novel xanthomonadin-dialkylresorcinol hybrid named arcuflavin was identified in Azoarcus sp. BH72 by a combination of feeding experiments, HPLC-MS and MALDI-MS and gene clusters encoding the biosynthesis of this non-isoprenoid aryl-polyene containing pigment are reported. A chorismate-utilizing enzyme from the XanB2-type producing 3- and 4-hydroxybenzoic acid and an AMP-ligase encoded by these gene clusters were characterized, that might perform the first two steps of the polyene biosynthesis. Furthermore, a detailed analysis of the already known or novel biosynthesis gene clusters involved in the biosynthesis of polyene containing pigments like arcuflavin, flexirubin and xanthomonadin revealed the presence of similar gene clusters in a wide range of bacterial taxa, suggesting that polyene and polyene-dialkylresorcinol pigments are more widespread than previously realized.
A revision of the genus Asterostegus Mortensen, 1933 (Echinodermata: Ophiuroidea: Euryalidae) is based on seven specimens, including the holotype of Asterostegus maini McKnight, 2003. A new species, Asterostegus sabineae sp. nov., is described from off Reunion Island and two other species, A. tuberculatus Mortensen, 1933 and A. maini, are redescribed. A tabular key to the three species of the genus Asterostegus is provided. Some terminology of the taxonomy of euryalid ophiuroids is revised.
The types of nominal species of Diapriinae in the collection of the Natural History Museum, London, are catalogued. Lectotypes are designated for the following taxa: Diapria peraffinis Ashmead, 1896; D. smithii Ashmead, 1896; Galesus bipunctatus Ashmead,1894; G. (G.) foersteri var. nigricornis Kieffer, 1911; G. sexpunctatus Ashmead, 1893; G. walkeri Kieffer, 1907; Idiotypa nigriceps Kieffer, 1909; I. nigriceps Kieffer, 1911; I. pallida Ashmead, 1893; I. pallida Ashmead in Riley, Ashmead & Howard, 1894; Paramesius angustipennis Kieffer, 1911; P. cameroni Kieffer, 1911; Phaenopria cameroni Kieffer, 1911; P. halterata Kieffer, 1911; P. magniclavata Ashmead, 1896; Tropidopsis clavata Ashmead, 1893; T. clavata Ashmead in Riley, Ashmead & Howard, 1894. New combinations are proposed: Aneuropria bifurcata comb. nov. for Mantara bifurcata Dodd, 1920; Basalys quadridens comb. nov. for Microgalesus quadridens Kieffer, 1912; Coptera cratocerus comb. nov. for Galesus cratocerus Cameron, 1912; Coptera sexpunctata comb. nov. for Galesus sexpunctatus Ashmead, 1893; Doliopria magniclavata comb. nov. for Phaenopria magniclavata Ashmead, 1896; Spilomicrus aterrimus comb. nov. for Hoplopria aterrima Dodd,
1920; Spilomicrus campbellanus comb. nov. for Antarctopria campbellana Yoshimoto, 1964; Spilomicrus coelopae comb. nov. for Antarctopria coelopae Early, 1978; Spilomicrus diomedeae comb. nov. for Antarctopria diomedeae Early, 1978; Spilomicrus helosciomyzae comb. nov. for Malvina helosciomyzae Early & Horning, 1978; Spilomicrus insulae comb. nov. for Malvina insulae Early, 1980; Spilomicrus latigaster comb. nov. for Antarctopria latigaster Brues in Tillyard, 1920; Spilomicrus punctatus comb. nov. for Malvina punctata Cameron, 1889; Spilomicrus rekohua comb. nov. for Antarctopria rekohua Early, 1978; Trichopria bouceki comb. nov. for Oxypria bouceki Masner, 1959; Trichopria nigriceps comb. nov. for Tropidopria nigriceps Ashmead in Riley, Ashmead & Howard, 1894; Trichopria nigriceps comb. nov. for Xyalopria nigriceps Kieffer, 1907; Trichopria spinosiceps comb. nov. for Acidopria spinosiceps Dodd, 1920; Trichopria walkeri comb. nov. for Diapria walkeri Dalla Torre, 1890. New replacement names are proposed: Coptera mosselensis nom. nov. for C. nigricornis Nixon, 1930 preocc.; Coptera pijiguaorum nom. nov. for C. sexpunctata Montilla & García, 2008 preocc.; Spilomicrus kozlovi nom. nov. for S. punctatus Kozlov, 1978 preocc.; Trichopria fluminis nom. nov. for T. nigriceps (Kieffer, 1907) preocc.; T. thermarum nom. nov. for T. nigriceps (Kieffer, 1913) preocc. New specific synonyms are proposed: Basalys cursitans (Kieffer, 1911) = B. pedisequa (Kieffer, 1911) syn. nov. (the former removed from synonymy with B. parvus Thomson, 1858); B. iphicla Nixon, 1980 = B. macroptera (Kieffer, 1911) syn. nov.; Coptera bipunctata (Ashmead in Riley, Ashmead & Howard, 1894) = C. sexpunctata (Ashmead, 1893) syn. nov.; Idiotypa nigriceps Kieffer, 1911 = I. nigriceps Kieffer, 1909 syn. nov.; I. pallida Ashmead in Riley, Ashmead & Howard, 1894 = I. pallida Ashmead, 1893 syn. nov.; Psilus nigricornis (Kieffer, 1911) = P. fuscipennis (Curtis, 1831) syn. nov.; P. walkeri (Kieffer, 1907) = P. fuscipennis (Curtis, 1831) syn. nov.; T. bouceki (Masner, 1959) = T. conotoma (Kieffer, 1911) syn. nov.; Trichopria halterata (Kieffer, 1911) = T. halterata (Kieffer, 1909) syn. nov. New generic synonyms are proposed: Antarctopria Brues in Tillyard, 1920 = Spilomicrus Westwood, 1832 syn. nov.; Malvina Cameron, 1889 = Spilomicrus Westwood, 1832 syn. nov.; Mantara Dodd, 1920 = Aneuropria Kieffer, 1905 syn. nov.; Microgalesus Kieffer, 1912 = Basalys Westwood, 1833 syn. nov.; Xyalopria Kieffer, 1907 = Trichopria Ashmead, 1893 syn. nov. (Xyalopria is removed from synonymy with Megaplastopria Ashmead, 1903). A brief account of some aspects of the history of these types is given.
Three fossil leafhopper inclusions from Eocene Baltic amber, representing three new extinct genera and species, are described and illustrated. Eomegophthalmus lithuaniensis gen. et sp. nov. is tentatively placed in Megophthalminae, although it may represent the stem group from which Megophthalminae, Ulopinae, and Membracidae arose. Xestocephalites balticus gen. et sp. nov. and Brevaphrodella nigra gen. et sp. nov. are placed in Aphrodinae: Xestocephalini based on the structure of the head, leg chaetotaxy, and male genital capsule. These new genera and species represent the oldest known representatives of their respective subfamilies and the latter is the oldest known brachypterous adult leafhopper.
The study of the Portuguese marine ichthyofauna has a long historical tradition, rooted back in the 18th Century. Here we present an annotated checklist of the marine fishes from Portuguese waters, including the area encompassed by the proposed extension of the Portuguese continental shelf and the Economic Exclusive Zone (EEZ). The list is based on historical literature records and taxon occurrence data obtained from natural history collections, together with new revisions and occurrences. It comprises a total of 1191 species, distributed among 3 superclasses, 4 classes, 42 orders, 212 families and 617 genera. If considering only the EEZ and present territorial waters, this list represents an increase of 230 species (27.8%) and of 238 species (29.0%), when compared to the information available in FishBase (2012) and in the last checklist of marine and estuarine fishes of Portugal (1993), respectively. The order Perciformes shows the highest diversity, with 54 families, 162 genera and 299 species. Stomiidae (80 species), Myctophidae (71 species) and Macrouridae (37 species) are the richest families. From the listed species, 734 are present off mainland Portugal, 857 off the Azores and 766 off Madeira. Within the limits of the examined area, three species are reported for the first time in mainland Portugal and twenty-nine records are identified as doubtful. A total of 133 species have been recorded from the extended Portuguese continental shelf (2 off mainland Portugal, 117 off the Azores and 14 off Madeira), two of which are common to the Azores and Madeira extensions. Biogeographically, the Atlantic group is the most important (548 species – 46.01%), followed by the Lusitanian group (256 species – 21.49%), the African group (71 species – 5.96%), the Boreal group (34 species – 2.85%), the Mediterranean group (31 species – 2.60%), the Macaronesian group (21 species – 1.76%), the Atlantic/African group (19 species – 1.60%) and the Mediterranean/African and the Arctic groups, each with only 1 species (0.08%). Regarding the preferences for vertical habitat, the demersal fishes are the most important group (305 species – 25.61%), followed by the mesopelagic group (228 species – 19.14%), the bathypelagic group (164 species – 13.77%), the benthopelagic group (147 species – 12.34%), the bathydemersal group (115 species – 9.66%), the reef-associated group (88 species – 7.39%), the pelagic group (74 species – 6.21%), the epipelagic group (58 species – 4.87%) and 1 species (0.08%) of the benthic group. The oceanic habitat is the best represented group comprising 446 species (37.45%), followed by the shelf group (199 species – 16.71%), the slope group (164 species – 13.77%), the inner shelf group (89 species – 7.47%), the coastal group (70 species – 5.88%), the outer shelf group (29 species – 2.43%) and the oceanic/shelf group (7 species – 0.59%).