DC 8: Mechanisms in neuropsychiatric-like behavioral phenotypes and brain connectivity in TSC

Project part of Work Package 2

Objectives: Using zebrafish as an animal model, the PhD candidate will test various compounds and potentially epistatic genes for modification of neuropsychiatric-like behavioral phenotypes in the Tsc2 mutant zebrafish (automated behavioral analysis using Zebrabox with automated data analysis). The compounds will be selected based on our previously obtained RNAseq data from brain tissue. In addition, compounds acting as new pharmacological inhibitors (University of Tübingen- M. Gehringer and the University of Basel – M. Wymann) or interfering with movel molecular targets (Université Paris-Cité – M.Pende, University Clinic of Essen – K. Thedieck) will also be tested. For compounds that modify behaviors, the brain morphology will be assessed by whole- mount immunofluorescence of brain markers in the intact brains (single-cell-resolution whole brain imaging using light-sheet microscopy) and then the activity of specific regions and the connectivity development will be examined using transgenic lines with Ca-sensitive fluorescent probe GcaMP or fluorescently-labelled specific neuronal populations, respectively (live time-lapse imaging of connectivity development using light-sheet microscopy).

Expected Results: List of molecular pathways that modify neuropsychiatric-like behaviours associated with TSC linked to brain morphology or connectivity alterations.

Planned secondments: 3 months at UniFR for proteomics analysis of TSC mutant zebrafish brains. 3 months at Topadur for testing the in vivo activity of putative mTOR inhibitors.