Cassani, Davide Ariberto Domenico; (2019) Mechanics and Architecture of the Cellular Actomyosin Cortex. Doctoral thesis (Ph.D), UCL (University College London).

Preview

Text Cassani_DADCassani_PhDThesis.pdf Download (157MB) | Preview

Abstract

Cell shape changes are key to cell physiology and underlie fundamental processes such as cell migration, cell division and tissue morphogenesis. Cell shape changes are precisely controlled by cell surface mechanics. One of the main determinant of cell surface mechanics is the actomyosin cortex, a thin network of actin, myosin and actin associated proteins underlying the plasma membrane. Cortex mechanical properties are not determined only by the cortical network composition but also by its nano-scale architecture. Although previous studies have shown a relationship between cortex organization and mechanics, which aspects of cortex architecture contribute to the regulation of cell surface mechanics remain largely unknown. This gap in understanding is mostly due to the small dimensions of the cortex, with a thickness close to the diffraction limit of light microscopy, which makes the investigation of cortical nanoscale organization challenging. In my PhD, I investigated cortex nano-scale organization and asked how cortex architecture affects cortex mechanics. In order to assess cortex mechanics, I established an AFM-based assay to measure cell cortex tension. By combining AFM measurements with an analysis on cortex thickness and targeted protein depletion, I demonstrated that cortical network organization directly affects cortex tension. I then further explored cortical organization focusing on nano-scale architecture. Due to the high density of the cortex, previous techniques, including super-resolution imaging, had failed to unveil actin filament organization. To assess this, I established a method to image actin filaments in the cortex using cryo electron tomography. I used isolated cellular blebs, which have been shown to repolymerise a cortex similar to that of intact cells and have dimensions compatible with cryo electron tomography. Actin cortex organization in blebs was analyzed using custom-made software and key parameters of cortical architecture were extracted. My investigation on cortex nano-scale architecture provides, to my knowledge, the first quantification of cortical actin organization and brings new insights into how cortex microscopic properties regulate the mesoscopic properties of cells.

Download activity - last month

Export as CSVJSONXML

Download activity - last 12 months

Export as XMLJSONCSV

Downloads by country - last 12 months

Export as JSONXMLCSV

Archive Staff Only

View Item