Abstract

The secretory route from the Golgi apparatus to the plasma membrane is one of the most important intracellular trafficking pathways. This pathway is essential for delivery of soluble proteins to the extracellular space as well as for integral membrane proteins to the plasma membrane. Despite its importance, this pathway is still poorly understood in neurons, which are polarised cells with two forms of soluble protein secretion (regulated and constitutive), and a complex subcellular organisation. We will use super-resolution imaging to map the secretory route of soluble protein in human iPSC derived neurons. We have already performed a number of screens in non-polarised immortalised cells which led to identifying a host of new proteins directly involved in Golgi apparatus to plasma membrane trafficking. We will determine whether neurons use similar secretory molecular machinery by testing the role of the proteins we have identified. In addition, we will use CRISPRi to abrogate secretory machinery in neurons, and compare single cell morphology and architecture in an unbiased manner, to determine the consequences of abnormal secretion on global neuronal organisation. In summary, we will characterise the secretory protein trafficking route in neurons. We will identify functional machinery that allows this process to happen, and we will work towards a comprehensive understanding on the effects of loss of this machinery.