Drosophila CG3303 is an essential endoribonuclease linked to TDP-43-mediated neurodegeneration

Endoribonucleases participate in almost every step of eukaryotic RNA metabolism, acting either as degradative or biosynthetic enzymes. We previously identified the founding member of the Eukaryotic EndoU ribonuclease family, whose components display unique biochemical features and are flexibly involved in important biological processes, such as ribosome biogenesis, tumorigenesis and viral replication. Here we report the discovery of the CG3303 gene product, which we named DendoU, as a novel family member in Drosophila. Functional characterisation revealed that DendoU is essential for Drosophila viability and nervous system activity. Pan-neuronal silencing of dendoU resulted in fly immature phenotypes, highly reduced lifespan and dramatic motor performance defects. Neuron-subtype selective silencing showed that DendoU is particularly important in cholinergic circuits. At the molecular level, we unveiled that DendoU is a positive regulator of the neurodegeneration-associated protein dTDP-43, whose downregulation recapitulates the ensemble of dendoU-dependent phenotypes. This interdisciplinary work, which comprehends in silico, in vitro and in vivo studies, unveils a relevant role for DendoU in Drosophila nervous system physio-pathology and highlights that DendoU-mediated neurotoxicity is, at least in part, contributed by dTDP-43 loss-of-function

WiFi Related Radiofrequency Electromagnetic Fields Promote Transposable Element Dysregulation and Genomic Instability in Drosophila melanogaster

Exposure to artificial radio frequency electromagnetic fields (RF-EMFs) has greatly increased in recent years, thus promoting a growing scientific and social interest in deepening the biological impact of EMFs on living organisms. The current legislation governing the exposure to RF-EMFs is based exclusively on their thermal effects, without considering the possible non-thermal adverse health effects from long term exposure to EMFs. In this study we investigated the biological non-thermal effects of low-level indoor exposure to RF-EMFs produced by WiFi wireless technologies, using Drosophila melanogaster as the model system. Flies were exposed to 2.4 GHz radiofrequency in a Transverse Electromagnetic (TEM) cell device to ensure homogenous controlled fields. Signals were continuously monitored during the experiments and regulated at non thermal levels. The results of this study demonstrate that WiFi electromagnetic radiation causes extensive heterochromatin decondensation and thus a general loss of transposable elements epigenetic silencing in both germinal and neural tissues. Moreover, our findings provide evidence that WiFi related radiofrequency electromagnetic fields can induce reactive oxygen species (ROS) accumulation, genomic instability, and behavioural abnormalities. Finally, we demonstrate that WiFi radiation can synergize with RasV12 to drive tumor progression and invasion. All together, these data indicate that radiofrequency radiation emitted from WiFi devices could exert genotoxic effects in Drosophila and set the stage to further explore the biological effects of WiFi electromagnetic radiation on living organisms

Stress, transposons and genome evolution

After Darwin's book on the origin of species by the natural selection, the theory of his precursor Lamarck was never completely abandoned. Over time, the observation of strange natural phenomena has occasionally resurrected the concept of the heredity of acquired characters. To explain, in Darwinian sense, some of the apparent Lamarckian-like phenomena, Waddington elaborated the “canalization and assimilation” concepts (Waddington, 1959). He observed that some phenotypic traits induced in Drosophila pupae by heat shock treatment and selected for a number of generations in the presence of the same stress, became heritable, thereby showing that an induced phenotypic trait could be inherited trough the germ line. Waddington hypothesized the existence of a cryptic genetic variation that is maintained hidden due to the robustness of the developmental process that he indicated as “canalization”. If an environmental stress is strong enough to overcome this robustness, the development pathway can change because of the expression of a cryptic genetic variant. Then, this variant can be selected and become heritable by an “assimilation” process. During the last few years, data supporting this view and providing possible molecular explanations were published. Rutheford and Lindquist (1998) showed that, in Drosophila, impairment of Hsp90 function induces morphogenetic variants that occasionally became fixed and stably transmitted. The interpretation was that Hsp90 is a capacitor of morphological evolution and buffers a pre-existing genetic variation that is not expressed and accumulates in neutral conditions. The stress sensitive storage and release of genetic variation by Hsp90 would favour adaptive evolution. However, our recent study has suggested a different explanation of these results (Specchia et al., 2010). It has been demonstrated that Hsp90 is involved in repression of transcription and mobilization of transposable elements in germ cells by affecting piRNA biogenesis. The reduction of HSP90 causes stress response-like activation and transposition of mobile elements along with a wide range of phenotypic variants due to the transposon insertions to the corresponding genes. On the basis of these observations, we have suggested that Hsp90, rather than functioning as a capacitor, acts, when absent, as a mutator, capable of causing activation and transposition of mobile elements through impairment of piRNAi silencing. Consequently, we propose that stress causes the activation of transposons that would induce de novo gene mutations, affecting development pathways; mutations can be expressed and fixed across subsequent generations by an assimilation process consisting of a co-selection of a somatic and a germinal event giving the same phenotype. This view implies that transposon activation is a major reaction of genomes to environmental stresses and represents a powerful adaptive response

Retrotransposon activation and genomic instability participate to Huntington Disease pathogenesis in a Drosophila melanogaster model

Huntington's disease (HD) is a late-onset, autosomal dominant disorder characterized by progressive motor dysfunction, early death and psychiatric disturbances. The disease is caused by a CAG repeat expansion in the IT15 gene, which elongates a stretch of polyglutamine (polyQ) at the amino-terminus of the HD protein, huntingtin (Htt). Despite the accumulated data on the molecular basis of neurodegeneration, no cure is still available. It is therefore important to keep investigating potential previously unnoticed pathways that may be altered in HD and target of therapeutic treatments. Transposable elements (TEs) are mobile genetic elements that constitute a large fraction of eukaryotic genomes. Retrotransposons represent approximately 40% and 30% of the human and Drosophila genomes. Mounting evidences suggest mammalian L1 elements are normally active during neurogenesis. Interestingly, recent reports show that unregulated activation of TE is associated with neurodegenerative diseases. Our experimental results show that retrotransposon transcripts are up-regulated in HD brain and that their inhibition determines the block of polyQ-dependent neurodegeneration. Moreover, we found a high rate of DNA damage and chromosomal abnormalities in HD brains. Taken together, these data suggest that TE activation and genomic instability represent two important pieces in the complicated puzzle of polyQ-induced neurotoxicity

Drosophila melanogaster as a model to study in vivo the functional role of Transposable Elements in Huntington’s Disease pathogenesis

Huntington's disease (HD) is a late-onset disorder characterized by progressive motor dysfunction, cognitive decline and psychiatric disturbances. The disease is caused by a CAG repeat expansion in the IT15 gene, which elongates a stretch of polyglutamine at the amino terminus of the huntingtin protein. Despite the impressive data that have been accumulated on the molecular basis of neurodegeneration, no cure is still available. It is therefore important to keep investigating potential previously unnoticed pathways that may be altered in HD and target of therapeutic treatments. Transposable elements (TEs) are mobile genetic elements that constitute a large fraction of eukaryotic genomes. Retrotransposons replicate through an RNA intermediate and represent approximately 40% and 30% of the human and Drosophila genomes. Mounting evidences suggest mammalian L1 elements are normally active during neurogenesis. Interestingly, recent reports show that unregulated activation of TEs is associated with neuropathology. Our experimental results obtained in Drosophila HD model, suggest that TE activation may represent an important piece in the complicated puzzle of polyQ-induced neurotoxicity

Canalization by selection of de novo-induced mutations

One of the most fascinating scientific problems, a subject of intense debate, is that of the mechanisms of biological evolution. In this context, Waddington elaborated the concepts of "canalization and assimilation" to explain how an apparently somatic variant induced by stress could become heritable through the germline in Drosophila. He resolved this seemingly Lamarckian phenomenon by positing the existence of cryptic mutations that can be expressed and selected under stress. To investigate the relevance of such mechanisms, we performed experiments following the Waddington procedure, then isolated and fixed three phenotypic variants along with another induced mutation that was not preceded by any phenocopy. All the fixed mutations we looked at were actually generated de novo by DNA deletions or transposon insertions, highlighting a novel mechanism for the assimilation process. Our study shows that heat-shock stress produces both phenotypic variants and germline mutations, and suggests an alternative explanation to that of Waddington for the apparent assimilation of an acquired character. The selection of the variants, under stress, for a number of generations allows for the co-selection of newly induced corresponding germline mutations, making the phenotypic variants appear heritable

WiFi Related Radiofrequency Electromagnetic Fields Promote Transposable Element Dysregulation and Genomic Instability in <i>Drosophila melanogaster</i>

Exposure to artificial radio frequency electromagnetic fields (RF-EMFs) has greatly increased in recent years, thus promoting a growing scientific and social interest in deepening the biological impact of EMFs on living organisms. The current legislation governing the exposure to RF-EMFs is based exclusively on their thermal effects, without considering the possible non-thermal adverse health effects from long term exposure to EMFs. In this study we investigated the biological non-thermal effects of low-level indoor exposure to RF-EMFs produced by WiFi wireless technologies, using Drosophila melanogaster as the model system. Flies were exposed to 2.4 GHz radiofrequency in a Transverse Electromagnetic (TEM) cell device to ensure homogenous controlled fields. Signals were continuously monitored during the experiments and regulated at non thermal levels. The results of this study demonstrate that WiFi electromagnetic radiation causes extensive heterochromatin decondensation and thus a general loss of transposable elements epigenetic silencing in both germinal and neural tissues. Moreover, our findings provide evidence that WiFi related radiofrequency electromagnetic fields can induce reactive oxygen species (ROS) accumulation, genomic instability, and behavioural abnormalities. Finally, we demonstrate that WiFi radiation can synergize with RasV12 to drive tumor progression and invasion. All together, these data indicate that radiofrequency radiation emitted from WiFi devices could exert genotoxic effects in Drosophila and set the stage to further explore the biological effects of WiFi electromagnetic radiation on living organisms