The efficient delivery of cellular constituents to their proper location is of fundamental importance for all cells and is of particular interest to neuroscientists, because of the unique functions and complex architecture of neurons. Protein Trafficking in Neurons examines mechanisms of protein trafficking and the role of trafficking in neuronal functioning from development to plasticity to disease. The book is divided into seven sections that review mechanisms of protein transport, the role of protein trafficking in synapse formation, exo- and endocytosis, transport of receptors, trafficking of ion channels and transporters, comparison of trafficking mechanisms in neuronal vs. non-neuronal cell types, and the relationship between trafficking and neuronal diseases such as Alzheimer's, Huntington's and Prion Diseases. - Provides a comprehensive examination of membrane/protein movement in neuronal function - Sections on synapse development, synaptic transmission, and the role of trafficking in neurological disease - Includes a focus on Molecular Mechanisms - Illustrated with color summary pictures - The only book examining protein trafficking and its functional implications, written by leaders in the field

This book covers the past, present and future of the intra-cellular trafficking field, which has made a quantum leap in the last few decades. It details how the field has developed and evolved as well as examines future directions.

The Neuronal Cytoskeleton, Motor Proteins, and Organelle Trafficking in the Axon, a new volume in the Methods in Cell Biology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in neuronal cells, and includes sections on such topics as actin transport in axons and neurofilament transport. - Covers an increasingly appreciated field in cell biology - Includes both established and new technologies - Contributed by experts in the field

Neurons are highly polarized cells with extremely long projections: dendrites and axons (together referred to as neurites). The central secretory dogma states that all protein synthesis and early secretory events take place in the cell body of a neuron, the soma. Yet, computational studies fail to reconcile experimental evidence with this notion proposing that there must be local protein synthesis and trafficking routes in distal neurites to support the function of a neuronal cell (reviewed in Chapter 1). Indeed, multiple lines of evidence from transcriptomics and protein translation studies indicate that local protein synthesis takes place independently of the soma. Amongst locally translated proteins, there are numerous secreted and membrane proteins. Yet, it is not clear how these newly synthesized proteins can be secreted locally, mainly, because the secretory pathway is largely understudied, especially in human neurons. Thus, I used human-induced pluripotent stem cells that I differentiated into cortical neurons to study the secretory pathway in distal neurites. In Chapter 2, I show that the secretory events occur in distal neuronal projections independently of the cell body. In fact, I find that nascent proteins are trafficked using an unconventional pathway via recycling endosomes. In addition, I uncover surprising new roles for TFG, a ubiquitously expressed protein and an important player in the early secretory pathway. Missense mutations in TFG have been identified in patients with neurodegenerative disorders though it is not clear how TFG results in disease. In Chapter 2, I show that TFG is not only important in the early secretory events in the conventional ER-to-Golgi trafficking, but it also mediates the unconventional secretory route where newly synthesized proteins exit ER and enter endosomes locally in distal neurites. Curiously, I find that TFG abundantly localizes with endosomes, suggesting that it must have an additional role in the endosomal pathway. Thus, I find that TFG has three distinct roles in neuronal cells: 1) conventional ER-to-Golgi trafficking in the soma, 2) unconventional ER-to-endosome route in distal neurites, 3) regulation of endosomal pathway. Future work, discussed in Chapter 3, will shed light on how TFG regulates the latter and how mutations in TFG lead to neurodegeneration.

Proper neuronal development and function requires precise sorting and delivery of various elements from the soma to the synapse. Important mediators of intracellular transport events are the actin-based class V myosin motors, which are involved in organelle transport in various cell types. Two myosin V family members, myosin Va and Vb, are present in the brain, however, the identity of cargoes transported by these motors is unknown. The objective of this thesis was to conduct molecular and cell biological studies to identify and characterize novel myosin V cargoes in neurons. The first approach I used was to characterize the distribution of candidate protein cargoes after blocking the function of endogenous myosin Va and Vb with dominant-negative (DN) versions. I found that in developing neurons, expression of DN myosin Vb, but not DN myosin Va, resulted in the accumulation in the soma of the AMPA-type glutamate receptor subunit, GluR1, and a reduction of its surface expression. I also found that myosin Vb-mediated trafficking of GluR1 required an interaction with the GTPase Rab11. These results reveal a novel mechanism for the transport of a specific glutamate receptor subunit mediated by myosin Vb and Rab11. As an alternative approach to identify myosin Va binding partners in the brain, we conducted a yeast-two hybrid screen of a rat brain cDNA library using the cargo binding domain of myosin Va. Among the proteins identified in our screen, I selected a protein of unknown function previously identified as Rab-lysosomal-interacting protein like 2 (RILPL2) and further assessed its function. I found that RILPL2 expression in non-neuronal cells resulted in morphological changes and activation of the Rho GTPase Rac1. In developing neurons, gain or loss of RILPL2 function altered the density of dendritic spine protrusions and increased phosphorylation of the Rac1 effector Pak. These findings uncover a novel role for the myosin Va-interacting protein, RILPL2, in regulati.

A do-it-yourself manual for culturing nerve cells, complete with recipes and protocols.