PUBLICATION

Presynaptic architecture of the larval zebrafish neuromuscular junction

Authors
Helmprobst, F., Frank, M., Stigloher, C.
ID
ZDB-PUB-150315-10
Date
2015
Source
The Journal of comparative neurology   523(13): 1984-97 (Journal)
Registered Authors
Stigloher, Christian
Keywords
electron microscopy, electron tomography, freeze substitution, high-pressure freezing, synapse
MeSH Terms
  • Age Factors
  • Animals
  • Freeze Fracturing
  • Imaging, Three-Dimensional
  • Larva/cytology*
  • Larva/ultrastructure
  • Microscopy, Electron
  • Models, Anatomic
  • Neuromuscular Junction/growth & development
  • Neuromuscular Junction/metabolism
  • Neuromuscular Junction/ultrastructure*
  • Presynaptic Terminals/metabolism
  • Presynaptic Terminals/ultrastructure*
  • Secretory Vesicles/ultrastructure
  • Zebrafish
PubMed
25766140 Full text @ J. Comp. Neurol.
Abstract
In this study we show the ultrastructural architecture of larval zebrafish (Danio rerio) neuromuscular junctions in three dimensions. We compare classical electron microscopy fixation techniques with high-pressure freezing followed by freeze substitution (HPF/FS) in combination with electron tomography. Further, we compare the structure of neuromuscular junctions in 4 and 8 dpf zebrafish larvae with HPF/FS as this allows for close-to-native ultrastructural preservation. We discovered that synaptic vesicles of 4 dpf zebrafish larvae are larger compared to 8 dpf larvae. Furthermore, we described two types of dense core vesicles and quantified a filamentous network of small filaments interconnecting synaptic vesicles as well as tethers connecting synaptic vesicles to the presynaptic cell membrane. In the center of active zones we found elaborate electron dense projections physically connecting vesicles of the synaptic vesicle pool to the presynaptic membrane. Altogether this study establishes the basis for systematic comparisons of synaptic architecture at high resolution in three dimensions of an intact vertebrate in a close-to-native state. Furthermore, we provide quantitative information that build the basis for diverse systems biology approaches in neuroscience, spanning from comparative anatomy to cellular simulations. This article is protected by copyright. All rights reserved.
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