A high-definition approach for quantitative experimental measurements of membrane protein mechanosensitivity

Current approaches to treat neurodegenerative diseases rely on invasively implanted electrodes to stimulate brain neurons. However, such treatments have shown high risks of infection or invasive damage. A high potential alternative is optogenetics, which combines optics with genetics, to manipulate neuronal activity. However, this method is unable to target deep brain regions. To address this issue, sonogenetics is a promising toolkit for targeted neuromodulation through acoustic fields. Sonogenetics, which focuses on modulation of sound sensitive neurons via the expression of mechanosensitive channel proteins, is non-invasive and reaches through intact thin bone and deep tissue. Recently, Fan et al. demonstrated the expression of ultrasound sensing protein mPrestin in dopaminergic neurons, resulting in a reduction in neurodegeneration. The exact mechanisms by which ultrasound stimulates neurons and mechanosensitive proteins remains poorly understood. Therefore, fundamental research is required to identify mechanosensitive proteins and investigate the mechanisms by which acoustic fields impact protein responses. Giant unilamellar vesicles (GUVs) are useful cell membrane models for investigating membrane-bound proteins that are selectively activated by sound. After integrating transmembrane proteins in GUV constructs, mechanosensitive proteins are artificially activated through acoustic energies and can generate spatiotemporally localized effects on demand. This project aims to develop a promising strategy for the investigation of mechanosensitive membrane proteins. In order to understand the fundamental mechanisms that activate mechanosensitive proteins, a testing platform is essential to gather useful experimental data. A microfluidic device is used to induce and measure expression of membrane-bound proteins, investigating the mechanisms underlying sonogenetic stimulation. These proteins are selectively stimulated via the application of acoustic fields, modulating neuron behaviour and thus making them a promising therapeutic toolkit to treat neurodegenerative diseases. Moreover, this work further improves our understanding of the optimal parameters for neural modulation.