Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The secretory pathway in neurons requires efficient targeting of cargos and regulatory proteins to their release sites. Tomosyn contributes to synapse function by regulating synaptic vesicle (SV) and dense-core vesicle (DCV) secretion. While there is large support for the presynaptic accumulation of tomosyn in fixed preparations, alternative subcellular locations have been suggested. Here we studied the dynamic distribution of tomosyn-1 (Stxbp5) and tomosyn-2 (Stxbp5l) in mouse hippocampal neurons and observed a mixed diffuse and punctate localization pattern of both isoforms. Tomosyn-1 accumulations were present in axons and dendrites. As expected, tomosyn-1 was expressed in about 75% of the presynaptic terminals. Interestingly, also bidirectional moving tomosyn-1 and -2 puncta were observed. Despite the lack of a membrane anchor these puncta co-migrated with synapsin and neuropeptide Y, markers for respectively SVs and DCVs. Genetic blockade of two known tomosyn interactions with synaptotagmin-1 and its cognate SNAREs did not abolish its vesicular co-migration, suggesting an interplay of protein interactions mediated by the WD40 and SNARE domains. We hypothesize that the vesicle-binding properties of tomosyns may control the delivery, pan-synaptic sharing and secretion of neuronal signaling molecules, exceeding its canonical role at the plasma membrane.

Original languageEnglish
Pages (from-to)e0180912
JournalPLoS ONE
Volume12
Issue number7
DOIs
Publication statusPublished - 2017

Fingerprint

SNARE Proteins
secretory granules
Secretory Vesicles
Neurons
Synaptotagmin I
neurons
secretion
Synapsins
neuropeptide Y
binding properties
Synaptic Vesicles
Secretory Pathway
Neuropeptide Y
dendrites
regulatory proteins
Presynaptic Terminals
Cell membranes
synapse
Dendrites
Anchors

Keywords

  • Animals
  • Axons
  • Binding Sites
  • Blotting, Western
  • Cells, Cultured
  • Dendrites
  • Hippocampus
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microscopy, Confocal
  • Microscopy, Immunoelectron
  • Nerve Tissue Proteins
  • Neurons
  • Neuropeptide Y
  • Presynaptic Terminals
  • Protein Binding
  • Protein Transport
  • R-SNARE Proteins
  • Secretory Vesicles
  • Synapsins
  • Synaptic Vesicles
  • Synaptotagmin I
  • Journal Article

Cite this

@article{3c0380d7265440048b2a3408730c69d0,
title = "Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains",
abstract = "The secretory pathway in neurons requires efficient targeting of cargos and regulatory proteins to their release sites. Tomosyn contributes to synapse function by regulating synaptic vesicle (SV) and dense-core vesicle (DCV) secretion. While there is large support for the presynaptic accumulation of tomosyn in fixed preparations, alternative subcellular locations have been suggested. Here we studied the dynamic distribution of tomosyn-1 (Stxbp5) and tomosyn-2 (Stxbp5l) in mouse hippocampal neurons and observed a mixed diffuse and punctate localization pattern of both isoforms. Tomosyn-1 accumulations were present in axons and dendrites. As expected, tomosyn-1 was expressed in about 75{\%} of the presynaptic terminals. Interestingly, also bidirectional moving tomosyn-1 and -2 puncta were observed. Despite the lack of a membrane anchor these puncta co-migrated with synapsin and neuropeptide Y, markers for respectively SVs and DCVs. Genetic blockade of two known tomosyn interactions with synaptotagmin-1 and its cognate SNAREs did not abolish its vesicular co-migration, suggesting an interplay of protein interactions mediated by the WD40 and SNARE domains. We hypothesize that the vesicle-binding properties of tomosyns may control the delivery, pan-synaptic sharing and secretion of neuronal signaling molecules, exceeding its canonical role at the plasma membrane.",
keywords = "Animals, Axons, Binding Sites, Blotting, Western, Cells, Cultured, Dendrites, Hippocampus, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Confocal, Microscopy, Immunoelectron, Nerve Tissue Proteins, Neurons, Neuropeptide Y, Presynaptic Terminals, Protein Binding, Protein Transport, R-SNARE Proteins, Secretory Vesicles, Synapsins, Synaptic Vesicles, Synaptotagmin I, Journal Article",
author = "Geerts, {Cornelia J} and Roberta Mancini and Ning Chen and Koopmans, {Frank T W} and Li, {Ka Wan} and Smit, {August B} and {van Weering}, {Jan R T} and Matthijs Verhage and Groffen, {Alexander J A}",
year = "2017",
doi = "10.1371/journal.pone.0180912",
language = "English",
volume = "12",
pages = "e0180912",
journal = "PLoS ONE",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "7",

}

Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains. / Geerts, Cornelia J; Mancini, Roberta; Chen, Ning; Koopmans, Frank T W; Li, Ka Wan; Smit, August B; van Weering, Jan R T; Verhage, Matthijs; Groffen, Alexander J A.

In: PLoS ONE, Vol. 12, No. 7, 2017, p. e0180912.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains

AU - Geerts, Cornelia J

AU - Mancini, Roberta

AU - Chen, Ning

AU - Koopmans, Frank T W

AU - Li, Ka Wan

AU - Smit, August B

AU - van Weering, Jan R T

AU - Verhage, Matthijs

AU - Groffen, Alexander J A

PY - 2017

Y1 - 2017

N2 - The secretory pathway in neurons requires efficient targeting of cargos and regulatory proteins to their release sites. Tomosyn contributes to synapse function by regulating synaptic vesicle (SV) and dense-core vesicle (DCV) secretion. While there is large support for the presynaptic accumulation of tomosyn in fixed preparations, alternative subcellular locations have been suggested. Here we studied the dynamic distribution of tomosyn-1 (Stxbp5) and tomosyn-2 (Stxbp5l) in mouse hippocampal neurons and observed a mixed diffuse and punctate localization pattern of both isoforms. Tomosyn-1 accumulations were present in axons and dendrites. As expected, tomosyn-1 was expressed in about 75% of the presynaptic terminals. Interestingly, also bidirectional moving tomosyn-1 and -2 puncta were observed. Despite the lack of a membrane anchor these puncta co-migrated with synapsin and neuropeptide Y, markers for respectively SVs and DCVs. Genetic blockade of two known tomosyn interactions with synaptotagmin-1 and its cognate SNAREs did not abolish its vesicular co-migration, suggesting an interplay of protein interactions mediated by the WD40 and SNARE domains. We hypothesize that the vesicle-binding properties of tomosyns may control the delivery, pan-synaptic sharing and secretion of neuronal signaling molecules, exceeding its canonical role at the plasma membrane.

AB - The secretory pathway in neurons requires efficient targeting of cargos and regulatory proteins to their release sites. Tomosyn contributes to synapse function by regulating synaptic vesicle (SV) and dense-core vesicle (DCV) secretion. While there is large support for the presynaptic accumulation of tomosyn in fixed preparations, alternative subcellular locations have been suggested. Here we studied the dynamic distribution of tomosyn-1 (Stxbp5) and tomosyn-2 (Stxbp5l) in mouse hippocampal neurons and observed a mixed diffuse and punctate localization pattern of both isoforms. Tomosyn-1 accumulations were present in axons and dendrites. As expected, tomosyn-1 was expressed in about 75% of the presynaptic terminals. Interestingly, also bidirectional moving tomosyn-1 and -2 puncta were observed. Despite the lack of a membrane anchor these puncta co-migrated with synapsin and neuropeptide Y, markers for respectively SVs and DCVs. Genetic blockade of two known tomosyn interactions with synaptotagmin-1 and its cognate SNAREs did not abolish its vesicular co-migration, suggesting an interplay of protein interactions mediated by the WD40 and SNARE domains. We hypothesize that the vesicle-binding properties of tomosyns may control the delivery, pan-synaptic sharing and secretion of neuronal signaling molecules, exceeding its canonical role at the plasma membrane.

KW - Animals

KW - Axons

KW - Binding Sites

KW - Blotting, Western

KW - Cells, Cultured

KW - Dendrites

KW - Hippocampus

KW - Mice, Inbred C57BL

KW - Mice, Knockout

KW - Microscopy, Confocal

KW - Microscopy, Immunoelectron

KW - Nerve Tissue Proteins

KW - Neurons

KW - Neuropeptide Y

KW - Presynaptic Terminals

KW - Protein Binding

KW - Protein Transport

KW - R-SNARE Proteins

KW - Secretory Vesicles

KW - Synapsins

KW - Synaptic Vesicles

KW - Synaptotagmin I

KW - Journal Article

U2 - 10.1371/journal.pone.0180912

DO - 10.1371/journal.pone.0180912

M3 - Article

VL - 12

SP - e0180912

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 7

ER -