Background: Synapse formation and maturation is arguably one of the most important steps to ensure the function of neural circuits during development. At the same time, synaptic defects and loss of synaptic functionality are common features across many neurodegenerative diseases. While modelling human neurons in vitro has been an invaluable tool for some time, studying single neurons in random arrangements does not allow to properly investigate the links between circuit architecture, connectivity and the molecular changes in synaptic machinery that occur at different stages of development, but also during the onset of neurodegeneration.
Our teams have recently developed a bioengineered platform that allows to generate complex large format neuronal circuity with controlled architecture, which can be used for live imaging studies but also can be disassembled in their constituent components to perform molecular studies (e.g. RNAseq/proteomics across different regions of the circuit). The aim of this project is to use this humanised model platform, together with live imaging reporters for synaptic activity and neuronal function, as well as engineered human stem cell lines, to characterise the links between circuit complexity and synaptic maturation.
Rotation Project:
During the rotation project we will optimise the protocols for assembly of complex neuronal circuitry with defined architecture using human stem cell derived cortical neurons, to obtain reliably different circuit arrangements (e.g. 2 nodes, 3 nodes, etc) and characterise the timeline of synaptic development by immunostaining across different timepoints
PhD Project:
During the PhD we will use the established protocols to first compare the timeline of synaptic establishment and maturation across different circuit architecture in vitro (Y1), then perform molecular analysis at the RNA and protein level by separating different compartment of the circuits across timepoints (Y2), and finally we will use this system to study synaptopathies mechanisms in neurodegenerative disorders (Y3).
Skills & Training: the student will work in an interdisciplinary team, learning different skills across neurobiology, stem cell differentiation, live imaging, electrophysiology, bioengineering, microfabrication and developmental biology.