Three-dimensional Analysis of Clathrin-mediated Membrane Trafficking in Living Cells

Clathrin-coated vesicles are the most prominent carriers of membrane traffic from cell surface to endosomes (endocytosis), a pathway by which hormones, transferrin, immunoglobulins, LDL, viruses, and their receptors enter cells. They are also important for traffic between endosomes and the trans-Golgi network. In this presentation, I will discuss (i) technological and analytical advances that I developed to directly visualize clathrin-mediated membrane traffic in three dimensions and in living cells; (ii) data obtained using these advances that defined a role for actin filament polymerization in counteracting membrane tension during clathrin-coated vesicle budding at the apical surface of polarized epithelial cells; and (iii) how these advances can be used to study a wide variety of biological processes that occur in living cells and tissues.