Formation of Golgi-Derived Active Zone Precursor Vesicles

Department of Psychiatry and Behavioral Sciences, Nancy Pritzker Laboratory, Stanford University, Palo Alto, California 94304-5485, USA.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience (Impact Factor: 6.34). 08/2012; 32(32):11095-108. DOI: 10.1523/JNEUROSCI.0195-12.2012
Source: PubMed


Vesicular trafficking of presynaptic and postsynaptic components is emerging as a general cellular mechanism for the delivery of scaffold proteins, ion channels, and receptors to nascent and mature synapses. However, the molecular mechanisms leading to the selection of cargos and their differential transport to subneuronal compartments are not well understood, in part because of the mixing of cargos at the plasma membrane and/or within endosomal compartments. In the present study, we have explored the cellular mechanisms of active zone precursor vesicle assembly at the Golgi in dissociated hippocampal neurons of Rattus norvegicus. Our studies show that Piccolo, Bassoon, and ELKS2/CAST exit the trans-Golgi network on a common vesicle that requires Piccolo and Bassoon for its proper assembly. In contrast, Munc13 and synaptic vesicle proteins use distinct sets of Golgi-derived transport vesicles, while RIM1α associates with vesicular membranes in a post-Golgi compartment. Furthermore, Piccolo and Bassoon are necessary for ELKS2/CAST to leave the Golgi in association with vesicles, and a core domain of Bassoon is sufficient to facilitate formation of these vesicles. While these findings support emerging principles regarding active zone differentiation, the cellular and molecular analyses reported here also indicate that the Piccolo-Bassoon transport vesicles leaving the Golgi may undergo further changes in protein composition before arriving at synaptic sites.

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    • "The Golgi apparatus also ensures the polarity of protein and lipid transport to axons and dendrites (Tang, 2001; Ramírez and Couve, 2011; Bonifacino, 2014). In particular, the Golgi apparatus is required for the assembly and axonal transport of synaptic vesicle precursors (Maas et al., 2012). Alterations of the Golgi apparatus could therefore be accompanied by loss or gain of function in protein sorting, processing and transport to the axons and synapses, leading to their degeneration. "
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    ABSTRACT: Pathological alterations of the Golgi apparatus, such as its fragmentation represent an early pre-clinical feature of many neurodegenerative diseases and have been widely studied in the motor neuron disease amyotrophic lateral sclerosis (ALS). Yet, the underlying molecular mechanisms have remained cryptic. In principle, Golgi fragmentation may result from defects in three major classes of proteins: structural Golgi proteins, cytoskeletal proteins and molecular motors, as well as proteins mediating transport to and through the Golgi. Here, we present the different mechanisms that may underlie Golgi fragmentation in animal and cellular models of ALS linked to mutations in SOD1, TARDBP (TDP-43), VAPB, and C9Orf72 and we propose a novel one based on findings in progressive motor neuronopathy (pmn) mice. These mice are mutated in the TBCE gene encoding the cis-Golgi localized tubulin-binding cofactor E, one of five chaperones that assist in tubulin folding and microtubule polymerization. Loss of TBCE leads to alterations in Golgi microtubules, which in turn impedes on the maintenance of the Golgi architecture. This is due to down-regulation of COPI coat components, dispersion of Golgi tethers and strong accumulation of ER-Golgi SNAREs. These effects are partially rescued by the GTPase ARF1 through recruitment of TBCE to the Golgi. We hypothesize that defects in COPI vesicles, microtubules and their interaction may also underlie Golgi fragmentation in human ALS linked to other mutations, spinal muscular atrophy (SMA), and related motor neuron diseases. We also discuss the functional relevance of pathological Golgi alterations, in particular their potential causative, contributory, or compensatory role in the degeneration of motor neuron cell bodies, axons and synapses.
    Full-text · Article · Dec 2015 · Frontiers in Neuroscience
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    • " through active transport ( Goldstein et al . , 2008 ; Hirokawa et al . , 2010 ; Figure 1A ) . It has been suggested that several AZ proteins , including bassoon , piccolo and ELKS , are trafficked as preassembled complexes , whereas SV proteins and a distinct set of AZ proteins are transported by other types of vesicles ( Shapira et al . , 2003 ; Maas et al . , 2012 ) . However , this notion has become questionable since SV proteins and AZ proteins were reported to be co - trafficked , likely via heterogeneous multi - vesicle transport complexes ( Tao - Cheng , 2007 ; Bury and Sabo , 2011 ; Wu et al . , 2013 ) . Due to the heterogeneity of these vesicles and potential co - trafficking of various ca"
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    ABSTRACT: The axon is the single long fiber that extends from the neuron and transmits electrical signals away from the cell body. The neuronal cytoskeleton, composed of microtubules (MTs), actin filaments and neurofilaments, is not only required for axon formation and axonal transport but also provides the structural basis for several specialized axonal structures, such as the axon initial segment (AIS) and presynaptic boutons. Emerging evidence suggest that the unique cytoskeleton organization in the axon is essential for its structure and integrity. In addition, the increasing number of neurodevelopmental and neurodegenerative diseases linked to defect in actin- and microtubule-dependent processes emphasizes the importance of a properly regulated cytoskeleton for normal axonal functioning. Here, we provide an overview of the current understanding of actin and microtubule organization within the axon and discuss models for the functional role of the cytoskeleton at specialized axonal structures.
    Full-text · Article · Aug 2015 · Frontiers in Molecular Neuroscience
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    • "In this overview, we will focus on the composition and mode of action of the presynaptic active zone. For specific aspects of the release sites, the reader is referred to recent reviews8910111213. The progress in the profiling of synaptosome proteomics have been reviewed in detail by Bai and Witzmann[14]. "

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