Presynaptic Rac1 controls synaptic strength through the regulation of synaptic vesicle priming

  1. Christian Keine
  2. Mohammed Al-Yaari
  3. Tamara Radulovic
  4. Connon I Thomas
  5. Paula Valino Ramos
  6. Debbie Guerrero-Given
  7. Mrinalini Ranjan
  8. Holger Taschenberger
  9. Naomi Kamasawa
  10. Samuel M Young Jr.  Is a corresponding author
  1. Carl von Ossietzky University of Oldenburg, Germany
  2. University of Iowa, United States
  3. Max Planck Florida Institute for Neuroscience, United States
  4. Göttingen Graduate School for Neurosciences, Biophysics, and Molecular Biosciences, Germany
  5. Max Planck Institute for Multidisciplinary Sciences, Germany

Abstract

Synapses contain a limited number of synaptic vesicles (SVs) that are released in response to action potentials (APs). Therefore, sustaining synaptic transmission over a wide range of AP firing rates and timescales depends on SV release and replenishment. Although actin dynamics impact synaptic transmission, how presynaptic regulators of actin signaling cascades control SV release and replenishment remains unresolved. Rac1, a Rho GTPase, regulates actin signaling cascades that control synaptogenesis, neuronal development, and postsynaptic function. However, the presynaptic role of Rac1 in regulating synaptic transmission is unclear. To unravel Rac1’s roles in controlling transmitter release, we performed selective presynaptic ablation of Rac1 at the mature mouse calyx of Held synapse. Loss of Rac1 increased synaptic strength, accelerated EPSC recovery after conditioning stimulus trains, and augmented spontaneous SV release with no change in presynaptic morphology or AZ ultrastructure. Analyses with constrained short-term plasticity models revealed faster SV priming kinetics and, depending on model assumptions, elevated SV release probability or higher abundance of tightly docked fusion-competent SVs in Rac1-deficient synapses. We conclude that presynaptic Rac1 is a key regulator of synaptic transmission and plasticity mainly by regulating the dynamics of SV priming and potentially SV release probability.

Data availability

All numerical data used to generate the figures are part of the respective source files. Experimental raw data and custom-written software central to the conclusion of this study are available at http://dx.doi.org/10.17632/c4b7gn8bh7 under the terms of the Creative Commons Attribution 4.0 License (CC BY 4.0).

Article and author information

Author details

  1. Christian Keine

    Department of Human Medicine, Carl von Ossietzky University of Oldenburg, Oldenburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Mohammed Al-Yaari

    Department of Anatomy and Cell Biology, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Tamara Radulovic

    Department of Human Medicine, Carl von Ossietzky University of Oldenburg, Oldenburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Connon I Thomas

    Electron Microscopy Core Facility, Max Planck Florida Institute for Neuroscience, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Paula Valino Ramos

    Department of Anatomy and Cell Biology, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Debbie Guerrero-Given

    Electron Microscopy Core Facility, Max Planck Florida Institute for Neuroscience, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Mrinalini Ranjan

    Göttingen Graduate School for Neurosciences, Biophysics, and Molecular Biosciences, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Holger Taschenberger

    Department of Molecular Neurobiology, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Naomi Kamasawa

    Electron Microscopy Core Facility, Max Planck Florida Institute for Neuroscience, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Samuel M Young Jr.

    Department of Anatomy and Cell Biology, University of Iowa, Iowa City, United States
    For correspondence
    samuel-m-young@uiowa.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7589-7612

Funding

National Institute on Deafness and Other Communication Disorders (R01 DC014093)

  • Samuel M Young Jr.

National Institute of Neurological Disorders and Stroke (R01 NS110742)

  • Samuel M Young Jr.

Deutsche Forschungsgemeinschaft (420075000)

  • Christian Keine

Max Planck Institute for Multidisciplinary Sciences (open access funding)

  • Mrinalini Ranjan
  • Holger Taschenberger

Max Planck Florida Institute (open access funding)

  • Connon I Thomas
  • Naomi Kamasawa

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All experiments were performed following animal welfare laws and approved by the Institutional Committee for Care and Use of Animals at the University of Iowa (PHS Assurance No. D16-00009 (A3021-01) (Animal Protocol 0021952) and complied with accepted ethical best practices.

Copyright

© 2022, Keine 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

  • 2,002
    views
  • 391
    downloads
  • 14
    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. Christian Keine
  2. Mohammed Al-Yaari
  3. Tamara Radulovic
  4. Connon I Thomas
  5. Paula Valino Ramos
  6. Debbie Guerrero-Given
  7. Mrinalini Ranjan
  8. Holger Taschenberger
  9. Naomi Kamasawa
  10. Samuel M Young Jr.
(2022)
Presynaptic Rac1 controls synaptic strength through the regulation of synaptic vesicle priming
eLife 11:e81505.
https://doi.org/10.7554/eLife.81505

Share this article

https://doi.org/10.7554/eLife.81505

Further reading

    1. Cell Biology
    Affiong Ika Oqua, Kin Chao ... Alejandra Tomas
    Research Article

    G protein-coupled receptors (GPCRs) are integral membrane proteins which closely interact with their plasma membrane lipid microenvironment. Cholesterol is a lipid enriched at the plasma membrane with pivotal roles in the control of membrane fluidity and maintenance of membrane microarchitecture, directly impacting on GPCR stability, dynamics, and function. Cholesterol extraction from pancreatic beta cells has previously been shown to disrupt the internalisation, clustering, and cAMP responses of the glucagon-like peptide-1 receptor (GLP-1R), a class B1 GPCR with key roles in the control of blood glucose levels via the potentiation of insulin secretion in beta cells and weight reduction via the modulation of brain appetite control centres. Here, we unveil the detrimental effect of a high cholesterol diet on GLP-1R-dependent glucoregulation in vivo, and the improvement in GLP-1R function that a reduction in cholesterol synthesis using simvastatin exerts in pancreatic islets. We next identify and map sites of cholesterol high occupancy and residence time on active vs inactive GLP-1Rs using coarse-grained molecular dynamics (cgMD) simulations, followed by a screen of key residues selected from these sites and detailed analyses of the effects of mutating one of these, Val229, to alanine on GLP-1R-cholesterol interactions, plasma membrane behaviours, clustering, trafficking and signalling in INS-1 832/3 rat pancreatic beta cells and primary mouse islets, unveiling an improved insulin secretion profile for the V229A mutant receptor. This study (1) highlights the role of cholesterol in regulating GLP-1R responses in vivo; (2) provides a detailed map of GLP-1R - cholesterol binding sites in model membranes; (3) validates their functional relevance in beta cells; and (4) highlights their potential as locations for the rational design of novel allosteric modulators with the capacity to fine-tune GLP-1R responses.

    1. Cell Biology
    2. Immunology and Inflammation
    Alejandro Rosell, Agata Adelajda Krygowska ... Esther Castellano Sanchez
    Research Article

    Macrophages are crucial in the body’s inflammatory response, with tightly regulated functions for optimal immune system performance. Our study reveals that the RAS–p110α signalling pathway, known for its involvement in various biological processes and tumourigenesis, regulates two vital aspects of the inflammatory response in macrophages: the initial monocyte movement and later-stage lysosomal function. Disrupting this pathway, either in a mouse model or through drug intervention, hampers the inflammatory response, leading to delayed resolution and the development of more severe acute inflammatory reactions in live models. This discovery uncovers a previously unknown role of the p110α isoform in immune regulation within macrophages, offering insight into the complex mechanisms governing their function during inflammation and opening new avenues for modulating inflammatory responses.