Jonathan Kuo Rose Hills
Clathrin-Mediated Endocytosis in 3D Cerebral Organoids
Clathrin-mediated endocytosis (CME) is a critical process in cells that maintains the balance of signaling and receptor molecules on the cellular surface. This balance is critical in the development of the human cerebral cortex, during which neuroepithelial stem cells must choose to divide or differentiate. These cell fate decisions are dependent on the distribution of membrane signaling molecules and receptors, distributions we hypothesize are regulated by CME. However, technical limitations have, until recently, made it nearly impossible to study the dynamics of CME in the highly polarized NECs of a neuroepithelium within live tissue.
Here, we seek to study CME in live tissues by culturing 3D cerebral organoids that capture the spatial organization and polarity of NECs during development. Using adaptive optics lattice light-sheet microscopy (AO-LLSM) and computational image analysis, we will observe the dynamics of CME in 3D, focusing on AP2, Dynamin2 and ArpC3, 3 key endocytic proteins that we have fluorescently tagged. With this project, we hope to shed new light on the role of CME in early neurodevelopment in live 3D tissue.