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Role of liposoluble hormones and maternal mRNAs of their cognate receptors in the epigenetic long-term modulation of zebrafish development. In teleost fish, as in other oviparous vertebrates, the ovulated oocytes contain large amounts of liposoluble hormones, such as steroid, thyroid and retinoid hormones, that are taken up from the maternal circulation and accumulated in the egg’s yolk. Aim of this research is to determine whether changes in steroid hormone concentrations or inactivation of their receptors could affect offspring development, thus demonstrating that these hormones are not only correlated with the maternal hormonal condition during the oogenesis, but are also biologically active throughout the first embryogenetic steps. To this end, we have knocked down, in the zebrafish, the mRNAs of the estrogen receptor, ers2a, and of the glucocorticoid receptor, gr, and prepared mutant lines for gr, mr and pr (ongoing) by means of CRISPR/Cas9 approach. Moreover, we have prepared a zebrafish transgenic line that works as a living biosensor model to dynamically trace glucocorticoid transcriptional activity during development and adult life in zebrafish. Molecular characterization of the zebrafish ambra1a, ambra1b and epg5 genes and analysis of silencing effects on zebrafish development and adult life. Since 2011 she has worked on the molecular characterization of zebrafish genes and proteins involved on autophagy: in particular ambra1a, ambra1b (positive regulator of this process) and epg5 (Rb7 effector for the fusion between autophagosome and lysosome). The corresponding genes have been knocked out by means of CRISPR/Cas9 approach. Characterization of the effects on development and adult life is actually ongoing. Silencing of these genes results on impairment of the autophagy process but also of other aspects of zebrafish physiology. In particular we are trying to understand the interplay between autophagy and sex determination as well as reproduction.

Analysis of steroid hormones and their cognate receptors functions during zebrafish development and adult life. The research will be based on the use of zebrafish CRISPR/Cas9 mutant lines for glucocorticoid (gr) receptor and mineralocorticoid (mr) receptors we have recently prepared. Analysis of ambra1a, ambra1b and epg5 functions on zebrafish development and the autophagy process. The research will be based on the use of zebrafish CRISPR/Cas9 mutant lines for these proteins. In particular we plan to analyse the interplay between autophagy and reproduction. In collaboration with Prof. Alessandro Alboresi (Group of Photosynthesis and Plant Biotechnology, Prof. Morosinotto), we are also preparing ambra1a and ambra1b mutant lines of Physcomitrella patens for the study of the roles played, in this plant model, by these proteins.