Treatment with THI, an inhibitor of sphingosine-1-phosphate lyase, modulates glycosphingolipid metabolism and results therapeutically effective in experimental models of Huntington's disease.

Huntington's disease (HD) is a fatal neurodegenerative disorder with no effective cure currently available. Over the past few years our research has shown that alterations in sphingolipid metabolism represent a critical determinant in HD pathogenesis. In particular, aberrant metabolism of sphingosine-1-phosphate (S1P) has been reported in multiple disease settings, including human postmortem brains from HD patients. In this study, we investigate the potential therapeutic effect of the inhibition of S1P degradative enzyme SGPL1, by the chronic administration of the 2-acetyl-5-tetrahydroxybutyl imidazole (THI) inhibitor. We show that THI mitigated motor dysfunctions in both mouse and fly models of HD. The compound evoked the activation of pro-survival pathways, normalized levels of brain-derived neurotrophic factor, preserved white matter integrity, and stimulated synaptic functions in HD mice. Metabolically, THI restored normal levels of hexosylceramides and stimulated the autophagic and lysosomal machinery, facilitating the reduction of nuclear inclusions of both wild-type and mutant huntingtin proteins.

Methods to Test Endocrine Disruption in Drosophila melanogaster.

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor chemicals (EDCs). These chemicals may alter the normal balance of endocrine systems and lead to adverse effects, as well as an increasing number of hormonal disorders in the human population or disturbed growth and reduced reproduction in the wildlife species. For some EDCs, there are documented health effects and restrictions on their use. However, for most of them, there is still no scientific evidence in this sense. In order to verify potential endocrine effects of a chemical in the full organism, we need to test it in appropriate model systems, as well as in the fruit fly, Drosophila melanogaster. Here we report detailed in vivo protocols to study endocrine disruption in Drosophila, addressing EDC effects on the fecundity/fertility, developmental timing, and lifespan of the fly. In the last few years, we used these Drosophila life traits to investigate the effects of exposure to 17-α-ethinylestradiol (EE2), bisphenol A (BPA), and bisphenol AF (BPA F). Altogether, these assays covered all Drosophila life stages and made it possible to evaluate endocrine disruption in all hormone-mediated processes. Fecundity/fertility and developmental timing assays were useful to measure the EDC impact on the fly reproductive performance and on developmental stages, respectively. Finally, the lifespan assay involved chronic EDC exposures to adults and measured their survivorship. However, these life traits can also be influenced by several experimental factors that had to be carefully controlled. So, in this work, we suggest a series of procedures we have optimized for the right outcome of these assays. These methods allow scientists to establish endocrine disruption for any EDC or for a mixture of different EDCs in Drosophila, although to identify the endocrine mechanism responsible for the effect, further essays could be needed.

Effects of the synthetic estrogen 17-α-ethinylestradiol on Drosophila melanogaster: Dose and gender dependence.

17-a-ethinylestradiol (EE2) belongs to the increasing list of Endocrine Disruptors Chemicals (EDCs), able to interfere with the endocrine system in both vertebrates and invertebrates. Regardless of its great dispersion in the environment, to date there is still little knowledge about its action mechanisms and harmful effects in invertebrates. To better evaluate its potential role in invertebrates, we used the model system Drosophila melanogaster, an insect in which the hormonal response has been widely described. The effects of EE2 in D.melanogaster adults have been evaluated by using life traits as well as molecular endpoints. It was found that EE2 significantly decreases survival and fertility in both sexes, with a higher effect in female flies, as well as affects the expression of the Ecdysone Receptor (EcR), Estrogen Related Receptor (ERR), Yolk protein2 (Yp2) and yolkless (yl) genes. In conclusion, our results suggest that EE2 treatment may have potential toxic and endocrine effects on Drosophila melanogaster adults of both sexes. In particular, our data provide an indication that, after EE2 treatment, two of the genes involved in the vitellogenesis process (yl and Yp2) are transcribed in adult males where are mostly silent, and suggest future studies forward their use as potential molecular markers to EDCs exposure in Drosophila male.