Drosophila TNF modulates tissue tension in the embryo to facilitate macrophage invasive migration

Migrating cells penetrate tissue barriers during development, inflammatory responses, and tumor metastasis. We study if migration in vivo in such three-dimensionally confined environments requires changes in the mechanical properties of the surrounding cells using embryonic Drosophila melanogaster hemocytes, also called macrophages, as a model. We find that macrophage invasion into the germband through transient separation of the apposing ectoderm and mesoderm requires cell deformations and reductions in apical tension in the ectoderm. Interestingly, the genetic pathway governing these mechanical shifts acts downstream of the only known tumor necrosis factor superfamily member in Drosophila, Eiger, and its receptor, Grindelwald. Eiger-Grindelwald signaling reduces levels of active Myosin in the germband ectodermal cortex through the localization of a Crumbs complex component, Patj (Pals-1-associated tight junction protein). We therefore elucidate a distinct molecular pathway that controls tissue tension and demonstrate the importance of such regulation for invasive migration in vivo

Avaliação do contributo da expressão genética na tolerância à desidratação em Rosa × hybrida cultivada em humidade relativa elevada

Cultivation of cut flowers with high postharvest longevity is of extreme importance for consumer’s satisfaction. A negative water balance is indicated as the most important factor reducing longevity. This problem is more severe in plants grown at high relative air humidity (i.e. RH > 85%) mainly due to a poor stomatal functioning associated to lower abscisic acid (ABA) concentrations, leading to higher water loss. In this paper it is analysed whether a sensitive cut rose cultivar to high RH show different gene expression related to ABA metabolism when grown at moderate (60%) or high (90%) RH. RNA was isolated from fully developed leaves, harvested 180 min after the beginning of the dark period (stomatal closing stimulus) and the relative gene expression was evaluated in genes involved in: biosynthesis (AAO3 and NCED1), oxidation (CYP707A1 and CYP707A3) and conjugation (UGT75B2) of ABA. The stomatal conductivity in the dark period was 5.6-fold higher in intact plants developed at high RH compared to moderate RH-grown plants, showing the reduced closure capacity of the stomata during darkness. After 4 h of leaflet desiccation, leaflets grown at high RH showed a relative water content (RWC) 69% lower than the moderate RH-grown leaflets, validating the sensitivity of K023 to high RH during growth. At last, from the five studied genes only the expression of CYP707A3 was significantly reduced by high RH, without significant differences found in the other genes. In conclusion, the sensitivity of the genotype K023 to high RH cannot be explained by the expression of the genes evaluated in this study, using leaves harvested 180 min after the beginning of the dark period (closing stimulus). We expect that after transferring the plants to the postharvest conditions, inducing water stress, a more contrasting gene expression among RH levels will appear.O cultivo de flores de corte com elevada longevidade pós-colheita é de extrema importância para a satisfação do consumidor. O stress hídrico é uma das principais causas de uma longevidade reduzida. Este problema é agravado em plantas produzidas em ambiente de humidade relativa elevada (i.e. HR > 85%) devido maioritariamente a disfunções estomáticas associadas a baixas concentrações de ácido abcísico (ABA), conduzindo a acentuadas perdas de água. Neste estudo é analisado se uma cultivar sensível à HR elevada apresenta diferente expressão de genes relacionados com o metabolismo do ABA quando cultivada em HR moderada (60%) ou elevada (90%). Isolou-se RNA de folhas completamente expandidas, colhidas 180 min depois do início do período noturno, e avaliou-se a expressão relativa de genes envolvidos nos processos de: biossíntese (AAO3 e NCED1), oxidação (CYP707A1 e CYP707A3) e conjugação (UGT75B2) do ABA. A condutividade estomática noturna revelou-se 5,6 vezes mais elevada em plantas intactas desenvolvidas em HR elevada comparativamente a plantas desenvolvidas em HR moderada, mostrando a reduzida capacidade de fecho dos estomas durante o período de obscuridade. Ao fim de 4 h de desidratação, folíolos cultivados em HR elevada apresentaram um conteúdo relativo em água (CRA) 69% inferior aos folíolos cultivados em HR moderada, comprovando-se a sensibilidade do genótipo K023 à elevada HR durante o crescimento. Por fim, dos cinco genes estudados apenas a expressão de CYP707A3 foi significativamente reduzida pela HR elevada, não se verificando diferenças significativas nos restantes genes. Concluiu-se que para o genótipo K023 a sensibilidade à HR elevada não pode ser explicada pela expressão dos genes avaliados neste estudo, recorrendo a folhas colhidas após 180 min de obscuridade (estímulo de fecho). Será de esperar que após a transferência das plantas para condições de pós-colheita, induzindo stress hídrico, a expressão genética entre níveis de HR será mais contrastante.info:eu-repo/semantics/publishedVersio

Evolutionary History of the Vertebrate Mitogen Activated Protein Kinases Family

Background: The mitogen activated protein kinases (MAPK) family pathway is implicated in diverse cellular processes and pathways essential to most organisms. Its evolution is conserved throughout the eukaryotic kingdoms. However, the detailed evolutionary history of the vertebrate MAPK family is largely unclear. Methodology/Principal Findings: The MAPK family members were collected from literatures or by searching the genomes of several vertebrates and invertebrates with the known MAPK sequences as queries. We found that vertebrates had significantly more MAPK family members than invertebrates, and the vertebrate MAPK family originated from 3 progenitors, suggesting that a burst of gene duplication events had occurred after the divergence of vertebrates from invertebrates. Conservation of evolutionary synteny was observed in the vertebrate MAPK subfamilies 4, 6, 7, and 11 to 14. Based on synteny and phylogenetic relationships, MAPK12 appeared to have arisen from a tandem duplication of MAPK11 and the MAPK13-MAPK14 gene unit was from a segmental duplication of the MAPK11-MAPK12 gene unit. Adaptive evolution analyses reveal that purifying selection drove the evolution of MAPK family, implying strong functional constraints of MAPK genes. Intriguingly, however, intron losses were specifically observed in the MAPK4 and MAPK7 genes, but not in their flanking genes, during the evolution from teleosts to amphibians and mammals. The specific occurrence of intron losses in the MAPK4 and MAPK7 subfamilies might be associated with adaptive evolution of the vertebrates by enhancing the gen

The INT6 Cancer Gene and MEK Signaling Pathways Converge during Zebrafish Development

BACKGROUND: Int-6 (integration site 6) was identified as an oncogene in a screen of tumorigenic mouse mammary tumor virus (MMTV) insertions. INT6 expression is altered in human cancers, but the precise role of disrupted INT6 in tumorigenesis remains unclear, and an animal model to study Int-6 physiological function has been lacking. PRINCIPAL FINDINGS: Here, we create an in vivo model of Int6 function in zebrafish, and through genetic and chemical-genetic approaches implicate Int6 as a tissue-specific modulator of MEK-ERK signaling. We find that Int6 is required for normal expression of MEK1 protein in human cells, and for Erk signaling in zebrafish embryos. Loss of either Int6 or Mek signaling causes defects in craniofacial development, and Int6 and Erk-signaling have overlapping domains of tissue expression. SIGNIFICANCE: Our results provide new insight into the physiological role of vertebrate Int6, and have implications for the treatment of human tumors displaying altered INT6 expression

The epigenetic regulator Histone Deacetylase 1 promotes transcription of a core neurogenic programme in zebrafish embryos

<p>Abstract</p> <p>Background</p> <p>The epigenetic regulator Histone Deacetylase 1 (Hdac1) is required for specification and patterning of neurones and myelinating glia during development of the vertebrate central nervous system (CNS). This co-ordinating function for Hdac1 is evolutionarily conserved in zebrafish and mouse, but the mechanism of action of Hdac1 in the developing CNS is not well-understood.</p> <p>Results</p> <p>A genome-wide comparative analysis of the transcriptomes of Hdac1-deficient and wild-type zebrafish embryos was performed, which identified an extensive programme of gene expression that is regulated by Hdac1 in the developing embryo. Using time-resolved expression profiling of embryos, we then identified a small subset of 54 genes within the Hdac1-regulated transcriptome that specifically exhibit robust and sustained Hdac1-dependent expression from early neurogenesis onwards. 18 of these 54 stringently Hdac1-regulated genes encode DNA-binding transcription factors that are implicated in promoting neuronal specification and CNS patterning, including the proneural bHLH proteins Ascl1a and Ascl1b, as well as Neurod4 and Neurod. Relatively few genes are strongly repressed by Hdac1 but expression of the Notch target gene <it>her6 </it>is attenuated by Hdac1 in specific sub-regions of the developing CNS, from early stages of neurogenesis onwards. Selected members of the stringently Hdac1-regulated group of genes were tested for Hdac1 binding to their promoter-proximal <it>cis</it>-regulatory elements. Surprisingly, we found that Hdac1 is specifically and stably associated with DNA sequences within the promoter region of <it>ascl1b </it>during neurogenesis, and that this Hdac1-<it>ascl1b </it>interaction is abolished in <it>hdac1 </it>mutant embryos.</p> <p>Conclusions</p> <p>We conclude that Hdac1 regulates histone acetylation and methylation in the developing zebrafish embryo and promotes the sustained, co-ordinate transcription of a small set of transcription factor genes that control expansion and diversification of cell fates within the developing CNS. Our <it>in vivo </it>chromatin immunoprecipitation results also suggest a specific function for Hdac1 in directly regulating transcription of a key member of this group of genes, <it>ascl1b</it>, from the beginning of neurogenesis onwards. Taken together, our observations indicate a novel role for Hdac1 as a positive regulator of gene transcription during development of the vertebrate CNS, in addition to its more well-established function in transcriptional repression.</p

High-Throughput Chemical Screen Identifies a Novel Potent Modulator of Cellular Circadian Rhythms and Reveals CKIα as a Clock Regulatory Kinase

A novel compound “longdaysin” was found to dramatically slow down the speed of the circadian clock through simultaneous inhibition of protein kinases CKIδ, CKIα, and ERK2

Is biotechnology (more) acceptable when it enables a reduction in phytosanitary treatments? A European comparison of the acceptability of transgenesis and cisgenesis

Reduced pesticide use is one of the reasons given by Europeans for accepting new genetic engineering techniques. According to the advocates of these techniques, consumers are likely to embrace the application of cisgenesis to apple trees. In order to verify the acceptability of these techniques, we estimate a Bayesian multilevel structural equation model, which takes into account the multidimensional nature of acceptability and individual, national, and European effects, using data from the Eurobarometer 2010 73.1 on science. The results underline the persistence of clear differences between European countries and whilst showing considerable defiance, a relatively wider acceptability of vertical gene transfer as a means of reducing phytosanitary treatments, compared to horizontal transfer

The Germinal Center Kinase GCK-1 Is a Negative Regulator of MAP Kinase Activation and Apoptosis in the C. elegans Germline

The germinal center kinases (GCK) constitute a large, highly conserved family of proteins that has been implicated in a wide variety of cellular processes including cell growth and proliferation, polarity, migration, and stress responses. Although diverse, these functions have been attributed to an evolutionarily conserved role for GCKs in the activation of ERK, JNK, and p38 MAP kinase pathways. In addition, multiple GCKs from different species promote apoptotic cell death. In contrast to these paradigms, we found that a C. elegans GCK, GCK-1, functions to inhibit MAP kinase activation and apoptosis in the C. elegans germline. In the absence of GCK-1, a specific MAP kinase isoform is ectopically activated and oocytes undergo abnormal development. Moreover, GCK-1- deficient animals display a significant increase in germ cell death. Our results suggest that individual germinal center kinases act in mechanistically distinct ways and that these functions are likely to depend on organ- and developmental-specific contexts