Determinants shaping the nanoscale architecture of the mouse rod outer segment

  1. Matthias Pöge
  2. Julia Mahamid
  3. Sanae S Imanishi
  4. Jürgen M Plitzko
  5. Krzysztof Palczewski  Is a corresponding author
  6. Wolfgang Baumeister  Is a corresponding author
  1. Max Planck Institute of Biochemistry, Germany
  2. European Molecular Biology Laboratory, Germany
  3. Indiana University, United States
  4. University of California, Irvine, United States

Abstract

The unique membrane organization of the rod outer segment (ROS), the specialized sensory cilium of rod photoreceptor cells, provides the foundation for phototransduction, the initial step in vision. ROS architecture is characterized by a stack of identically shaped and tightly packed membrane disks loaded with the visual receptor rhodopsin. A wide range of genetic aberrations have been reported to compromise ROS ultrastructure, impairing photoreceptor viability and function. Yet, the structural basis giving rise to the remarkably precise arrangement of ROS membrane stacks and the molecular mechanisms underlying genetically inherited diseases remain elusive. Here, cryo-electron tomography (cryo-ET) performed on native ROS at molecular resolution provides insights into key structural determinants of ROS membrane architecture. Our data confirm the existence of two previously observed molecular connectors/spacers which likely contribute to the nanometer-scale precise stacking of the ROS disks. We further provide evidence that the extreme radius of curvature at the disk rims is enforced by a continuous supramolecular assembly composed of peripherin-2 (PRPH2) and rod outer segment membrane protein 1 (ROM1) oligomers. We suggest that together these molecular assemblies constitute the structural basis of the highly specialized ROS functional architecture. Our Cryo-ET data provide novel quantitative and structural information on the molecular architecture in ROS and substantiate previous results on proposed mechanisms underlying pathologies of certain PRPH2 mutations leading to blindness.

Data availability

The subvolume averages of ROS disk rims are deposited in EMDB under the accession codes: EMD-13321, EMD-13322, EMD-13323 and EMD-13324.Two representative 4x binned tomograms for each acquisition scheme or mouse strain were deposited in EMPIAR under the accession codes: EMPIAR-10771, EMPIAR-10772 and EMPIAR-10773

Article and author information

Author details

  1. Matthias Pöge

    Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2331-4638
  2. Julia Mahamid

    European Molecular Biology Laboratory, Heidelberg, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6968-041X
  3. Sanae S Imanishi

    Department of Ophthalmology, Indiana University, Indianapolis, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9884-2123
  4. Jürgen M Plitzko

    Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6402-8315
  5. Krzysztof Palczewski

    Gavin Herbert Eye Institute, University of California, Irvine, Irvine, United States
    For correspondence
    kpalczew@uci.edu
    Competing interests
    Krzysztof Palczewski, is Chief Scientific Officer of Polgenix Inc..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0788-545X
  6. Wolfgang Baumeister

    Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
    For correspondence
    baumeist@biochem.mpg.de
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8154-8809

Funding

National Institutes of Health (R01EY030873)

  • Krzysztof Palczewski

National Institutes of Health (R01EY030912)

  • Krzysztof Palczewski

Max Planck Institute for Dynamics of Complex Technical Systems Magdeburg (Open-access funding)

  • Matthias Pöge

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

Ethics

Animal experimentation: The animals used were bred for scientific purposes. At the University of California, Irvine, all mice were housed in the vivarium where they were maintained on a normal mouse chow diet and a 12 h / 12 h light / dark cycle. All animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC, AUP-21-096) of the University of California, Irvine, and were conducted in accordance with the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Visual Research. The research at the Max Planck Institute of Biochemistry did not involve experiments on an animal. All animals were sacrificed prior to the removal of organs in accordance with the European Commission Recommendations for the euthanasia of experimental animals (Part 1 and Part 2). Breeding and housing as well as the euthanasia of the animals are fully compliant with all German (e.g. German Animal Welfare Act) and EC (e.g. Directive 86/609/EEC) applicable laws and regulations concerning care and use of laboratory animals. The Max Planck Institute of Biochemistry has a licence for breeding and housing of Iaboratory animals which includes the killing of animals solely for the use of their organs or tissues (No.4.3.2-5682/MPI für Biochemie - rural districts office).

Reviewing Editor

  1. Giulia Zanetti, Institute of Structural and Molecular Biology, Birkbeck, University of London, United Kingdom

Publication history

  1. Received: August 5, 2021
  2. Accepted: December 9, 2021
  3. Accepted Manuscript published: December 21, 2021 (version 1)
  4. Version of Record published: January 13, 2022 (version 2)
  5. Version of Record updated: January 17, 2022 (version 3)

Copyright

© 2021, Pöge 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.

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  1. Matthias Pöge
  2. Julia Mahamid
  3. Sanae S Imanishi
  4. Jürgen M Plitzko
  5. Krzysztof Palczewski
  6. Wolfgang Baumeister
(2021)
Determinants shaping the nanoscale architecture of the mouse rod outer segment
eLife 10:e72817.
https://doi.org/10.7554/eLife.72817
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