1. Neuroscience

Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis

  1. Mahdokht Kohansal-Nodehi
  2. John JE Chua
  3. Henning Urlaub
  4. Reinhard Jahn
  5. Dominika Czernik  Is a corresponding author
  1. Max Planck Institute for Biophysical Chemistry, Germany
  2. National University of Singapore, Singapore
  3. Max-Planck-Institute for Biophysical Chemistry, Germany
https://doi.org/10.7554/eLife.14530
Share this article
Cite this article
  1. Mahdokht Kohansal-Nodehi
  2. John JE Chua
  3. Henning Urlaub
  4. Reinhard Jahn
  5. Dominika Czernik
(2016)
Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
eLife 5:e14530.
https://doi.org/10.7554/eLife.14530
  2. Download BibTeX
  3. Download .RIS

Abstract

Neurotransmitter release is mediated by the fast, calcium-triggered fusion of synaptic vesicles with the presynaptic plasma membrane, followed by endocytosis and recycling of the membrane of synaptic vesicles. While many of the proteins governing these processes are known, their regulation is only beginning to be understood. Here we have applied quantitative phosphoproteomics to identify changes in phosphorylation status of presynaptic proteins in resting and stimulated nerve terminals isolated from the brains of Wistar rats. Using rigorous quantification, we identified 252 phosphosites that are either up- or downregulated upon triggering calcium-dependent exocytosis. Particularly pronounced were regulated changes of phosphosites within protein constituents of the presynaptic active zone, including bassoon, piccolo, and RIM1. Additionally, we have mapped kinases and phosphatases that are activated upon stimulation. Overall, our study provides a snapshot of phosphorylation changes associated with presynaptic activity and provides a foundation for further functional analysis of key phosphosites involved in presynaptic plasticity.

Article and author information

Author details

  1. Mahdokht Kohansal-Nodehi

    Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    Competing interests
    No competing interests declared.

  2. John JE Chua

    Interactomics and Intracellular Trafficking laboratory, Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore
    Competing interests
    No competing interests declared.

  3. Henning Urlaub

    Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    Competing interests
    No competing interests declared.

  4. Reinhard Jahn

    Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    Competing interests
    Reinhard Jahn, Reviewing editor, eLife.

  5. Dominika Czernik

    Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany
    For correspondence
    dominika.czernik@mpibpc.mpg.de
    Competing interests
    No competing interests declared.

Ethics

Animal experimentation: All animal procedures used here fully comply with the guidelines as stipulated in the section 4 of the Animal Welfare Law of the Federal Republic of Germany (section 4 of TierSchG, Tierschutzgesetz der Bundesrepublik Deutschland). All procedures were performed in the animal facility at the Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany registered accordingly to the section 11 Abs. 1 TierSchG as documented by 39 20 00_2a Si/rö, dated 11th Dec 2013 ("Erlaubnis, Wirbeltiere zur Versuchszwecken zu züchten und zu halten"), by the Veterinär- und Verbraucherschutzamt für den Landkreis und die Stadt Göttingen and examined regularly by the supervisory veterinary authority of the Landkreis Göttingen. All procedures were supervised by the animal welfare officer and the animal welfare committee of the Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany established accordingly to the TierSchG and the regulation about animal used in experiments, dated on 1st Aug 2013 (TierSchVersV, Tierschutz-Versuchstier-Verordung).

Copyright

© 2016, Kohansal-Nodehi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,546
    views
  • 1,006
    downloads
  • 44
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Mahdokht Kohansal-Nodehi
  2. John JE Chua
  3. Henning Urlaub
  4. Reinhard Jahn
  5. Dominika Czernik
(2016)
Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
eLife 5:e14530.
https://doi.org/10.7554/eLife.14530

Share this article

https://doi.org/10.7554/eLife.14530

Categories and tags

Research organism

Further reading

    1. Neuroscience

    Parabrachial CGRP neurons modulate active defensive behavior under a naturalistic threat

    Gyeong Hee Pyeon, Hyewon Cho ... Yong Sang Jo
    Research Article Updated

    Recent studies suggest that calcitonin gene-related peptide (CGRP) neurons in the parabrachial nucleus (PBN) represent aversive information and signal a general alarm to the forebrain. If CGRP neurons serve as a true general alarm, their activation would modulate both passive nad active defensive behaviors depending on the magnitude and context of the threat. However, most prior research has focused on the role of CGRP neurons in passive freezing responses, with limited exploration of their involvement in active defensive behaviors. To address this, we examined the role of CGRP neurons in active defensive behavior using a predator-like robot programmed to chase mice. Our electrophysiological results revealed that CGRP neurons encode the intensity of aversive stimuli through variations in firing durations and amplitudes. Optogenetic activation of CGRP neurons during robot chasing elevated flight responses in both conditioning and retention tests, presumably by amplifying the perception of the threat as more imminent and dangerous. In contrast, animals with inactivated CGRP neurons exhibited reduced flight responses, even when the robot was programmed to appear highly threatening during conditioning. These findings expand the understanding of CGRP neurons in the PBN as a critical alarm system, capable of dynamically regulating active defensive behaviors by amplifying threat perception, and ensuring adaptive responses to varying levels of danger.

    1. Evolutionary Biology
    2. Neuroscience

    Vision: The inner workings of a miniature eye

    Gregor Belušič
    Insight

    The first complete 3D reconstruction of the compound eye of a minute wasp species sheds light on the nuts and bolts of size reduction.

    1. Neuroscience

    Cortical tracking of hierarchical rhythms orchestrates the multisensory processing of biological motion

    Li Shen, Shuo Li ... Yi Jiang
    Research Article

    When observing others’ behaviors, we continuously integrate their movements with the corresponding sounds to enhance perception and develop adaptive responses. However, how the human brain integrates these complex audiovisual cues based on their natural temporal correspondence remains unclear. Using electroencephalogram (EEG), we demonstrated that rhythmic cortical activity tracked the hierarchical rhythmic structures in audiovisually congruent human walking movements and footstep sounds. Remarkably, the cortical tracking effects exhibit distinct multisensory integration modes at two temporal scales: an additive mode in a lower-order, narrower temporal integration window (step cycle) and a super-additive enhancement in a higher-order, broader temporal window (gait cycle). Furthermore, while neural responses at the lower-order timescale reflect a domain-general audiovisual integration process, cortical tracking at the higher-order timescale is exclusively engaged in the integration of biological motion cues. In addition, only this higher-order, domain-specific cortical tracking effect correlates with individuals’ autistic traits, highlighting its potential as a neural marker for autism spectrum disorder. These findings unveil the multifaceted mechanism whereby rhythmic cortical activity supports the multisensory integration of human motion, shedding light on how neural coding of hierarchical temporal structures orchestrates the processing of complex, natural stimuli across multiple timescales.