1,720,966 research outputs found
Chromatin Preparation and Chromatin Immunoprecipitation from Drosophila Heads
No full textChromatin immunoprecipitation (ChIP) is a widely used method to map protein-DNA interactions in vivo. Formaldehyde cross-linked chromatin is fragmented, and the protein of interest is immunoprecipitated using a specific antibody. The co-immunoprecipitated DNA is then purified and analyzed by quantitative PCR (ChIP-qPCR) or next-generation sequencing (ChIP-seq). Therefore, from the amount of DNA recovered, it can be inferred the localization and abundance of the target protein at specific loci or throughout the entire genome. This protocol describes how to perform ChIP from Drosophila adult fly heads
The role of transposable elements activity in aging and their possible involvement in laminopathic diseases
Full text linkEukaryotic genomes contain a large number of transposable elements, part of which are still active and able to transpose in the host genome. Mobile element activation is repressed to avoid deleterious effects, such as gene mutations or chromosome rearrangements. Control of transposable elements includes a variety of mechanisms comprising silencing pathways, which are based on the production of small non-coding RNAs. Silencing can occur either through transposable element RNA degradation or through the targeting of DNA sequences by heterochromatin formation and consequent transcriptional inhibition. Since the important role of the heterochromatin silencing, the gradual loss of heterochromatin marks in constitutive heterochromatin regions during the aging process promotes derepression of transposable elements, which is considered a cause of the progressive increase in genomic instability and of the activation of inflammatory responses. This review provides an overview of the effects of heterochromatin loss on the activity of transposable elements during the aging process and the possible impact on genome function. In this context, we discuss the possible role of the nuclear lamina, a major player in heterochromatin dynamics, in the regulation of transposable element activity and potential implications in laminopathic diseases
Spatial activation and repression of the drosophila vitelline membrane gene VM32E are switched by a complex cis-regulatory system
Full text linkComparative expression profiling of wild type Drosophila Malpighian tubules and von Hippel-Lindau haploinsufficient mutant
Full text linkThe von-Hippel Lindau (VHL) disease is a hereditary genetic disorder that predisposes to the onset of several highly vascularized benign and malignant tumors, developing with elevate frequency in the central nervous system and kidneys. The most-aggressive VHL tumor is ccRCC, the clear-cell renal cell carcinoma, affecting the kidney. VHL disease etiology can be attributed to the inheritance of a VHL loss-of-function allele, typically a deletion (Gnarra et al., 1994; Herman et al., 1994); this facilitates the somatic inactivation of the other allele (through amorphic mutations or gene silencing through promoter methylation), leading to the onset of the tumorous phenotype (Latif et al., 1993). This reveals the haploinsufficient behavior of the VHL gene.
The high vascularization of VHL tumors can be explained considering that human VHL protein is the substrate-binding subunit of an E3 ubiquitin ligase (Lonergan et al., 1998; Iwai et al., 1999; Kamura et al., 1999) involved in the poly-ubiquitination of HIF-1α transcription factor. This post-translational modification leads HIF-1α to proteosomal degradation (Maxwell et al., 1999). Loss of VHL function causes the stabilization of HIF-1α, triggering cellular response and adaptation to hypoxic conditions (expression of genes involved in glycolysis, angiogenesis and erythropoiesis) (Bader and Hsu, 2012). While this represents the canonical function of VHL, other HIF-1α-independent function of VHL have been identified, thanks to the contribution of model organisms (Hsu, 2012). Indeed, VHL gene function is conserved and also Drosophila has a VHL homolog, the dVHL gene (Adryan et al., 2000; Aso et al., 2000). dVHL is involved in the development of Drosophila vascular system (Adryan et al., 2000; Hsouna et al., 2010) and in morphogenesis of follicular epithelium of the egg chamber (Duchi et al., 2010). Interestingly, some VHL functions are mediated by Awd, an endocytic mediator whose human orthologs are NME1/2 metastasis suppressors (Rosengard et al., 1989). Awd is broadly required during Drosophila development since it is involved in epithelial morphogenesis (Nallamothu et al., 2008; Woolworth et al., 2009; Ignesti et al., 2014) and required for maintaining genomic stability (Romani et al., 2017). Moreover, Awd is also present into the extracellular fluids of Drosophila larvae (Romani et al., 2016, 2018).
In Drosophila, two pairs of monolayered epithelial Malpighian tubules, each composed of 100-150 cells, absolve to osmoregulation and excretion functions (Denholm and Skaer, 2009). Transcriptomic analysis of Malpighian tubules revealed that among genes that are here enriched there are homologs of human genes implicated into renal pathologies (Wang et al., 2004). This justifies the use of Drosophila Malpighian tubules as model system to gain insights into pathophysiology of human kidneys (Dow and Romero, 2010; Miller et al., 2013).
The dVHL1.1 allele is a loss of function mutation of the dVHL locus (Duchi et al., 2010; Hsouna et al., 2010). dVHL1.1/+ flies mimic the genetic condition of VHL patients. We carried out a genome-wide gene expression profiling of whole Malpighian tubules dissected from Drosophila females both heterozygous for the dVHL1.1 mutation and with two wild type copies of the dVHL gene. The comparison of differentially expressed genes in the two genetic backgrounds potentially allows to identify genes that are sensible to dVHL functional copy number. Quality control assessments of the data were performed and results obtained from the differential expression analysis were confirmed by qRT-PCR. With this approach we aimed to provide a well-controlled dataset for a better understanding of the VHL disease. Indeed, even if further molecular and functional characterization are needed, human homologs of the differentially expressed genes, if existing, could have a role in the somatic inactivation of the wild type copy of VHL and/or into the very first phase of cancer onset
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
The impact of drosophila awd/nme1/2 levels on notch and wg signaling pathways
Full text linkAwd, the Drosophila homologue of NME1/2 metastasis suppressors, plays key roles in many signaling pathways. Mosaic analysis of the null awdJ2A4 allele showed that loss of awd gene function blocks Notch signaling and the expression of its target genes including the Wingless (Wg/Wnt1) morphogen. We also showed that RNA interference (RNAi)-mediated awd silencing (awdi) in larval wing disc leads to chromosomal instability (CIN) and to Jun amino-terminal kinases (JNK)-mediated cell death. Here we show that this cell death is independent of p53 activity. Based on our previous finding showing that forced survival of awdi-CIN cells leads to aneuploidy without the hyperproliferative effect, we investigated the Wg expression in awdi wing disc cells. Interestingly, the Wg protein is expressed in its correct dorso-ventral domain but shows an altered cellular distribution which impairs its signaling. Further, we show that RNAi-mediated knock down of awd in wing discs does not affect Notch signaling. Thus, our analysis of the hypomorphic phenotype arising from awd downregulation uncovers a dose-dependent effect of Awd in Notch and Wg signaling
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Specific domains drive VM32E protein distribution and integration in Drosophila eggshell layers
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