Project description:The objective of this project is to study the proteome in different tissues of huntingtin knock-in allelic series in mice. In the current part of the study, heart samples from the complete allelic series collected at 2, 6, and 10 months were analyzed. To this end, comprehensive quantitative, label-free proteome analysis was performed using Evotec’s quantitative Deep Proteome Profiling technology.
Project description:Pregnant C57Bl6N mice were treated with 0 (corn oil), 1.5, 3.0, or 6.0 ug/kg TCDD on gd14.5. Fetal hearts were collected on gd17.5. Hearts from each litter were pooled onto one chip. 4 replicates of each condition were run on affymetrix MG_U74Av2 chips, using standard affymetrix protocols and controls. Keywords: dose-response, 3 doses plus corn oil control, 4 replicates
Project description:The zebrafish has the capacity to regenerate its heart after severe injury. While the function of a few genes during this process has been studied, we are far from fully understanding how genes interact to coordinate heart regeneration. To enable systematic insights into this phenomenon, we generated and integrated a dynamic co-expression network of heart regeneration in the zebrafish and linked systems-level properties to the underlying molecular events. Across multiple post-injury time points, the network displays topological attributes of biological relevance. We show that regeneration steps are mediated by modules of transcriptionally coordinated genes, and by genes acting as network hubs. We also established direct associations between hubs and validated drivers of heart regeneration with murine and human orthologs. The resulting models and interactive analysis tools are available at http://infused.vital-it.ch. Using a worked example, we demonstrate the usefulness of this unique open resource for hypothesis generation and in silico screening for genes involved in heart regeneration.
Project description:The transition from maternal to zygotic gene expression during zygotic genome activation (ZGA) is tightly associated with chromatin accessibility modulated by maternal transcription factors. However, due to technical limitations, it remains elusive how the chromatin regulatory landscape is established in Xenopus tropicalis ZGA, and let alone DNA binding transcription regulators involved in this process. Here, by developing concanavalin A beads-based nucleus capture followed by Tn5-mediated accessible chromatin assay with sequencing (CANTAC-seq), we generated a first genome-wide map of accessible chromatin in X. tropicalis embryos from early blastula to neurula stage. We found that open chromatin landscape is progressively established at cis-regulatory elements during ZGA. Based on the motif analysis and perturbation experiments, we demostrated E2f1, a well-known transcriptional activator, maintains a repressive chromatin environment preceding minor ZGA. Moreover, we identified another maternal factor Otx1 counteracts the inhibitory function of E2f1. Mechanistically, E2f1 and Otx1 co-regulate a set of genes required for zygotic gene transcription and germ layer differentiation before the midblastula transition (MBT) through a seesaw model. Taken together, our data reveals the dynamic chromatin regulatory landscape that accompanies early development and elaborates on the function of a coupled transcriptional repressor-activator regulating zygotic gene transcription in a classical model organism.