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- EMDB-11147: Heterozygous Opa1 knockout mouse photoreceptor inner segment mito... -

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Basic information

Entry
Database: EMDB / ID: EMD-11147
TitleHeterozygous Opa1 knockout mouse photoreceptor inner segment mitochondria
Map dataHeterozygous Opa1 knockout mouse photoreceptor inner segment mitochondria
Sample
  • Cell: Mouse retina
Biological speciesMus musculus (house mouse)
Methodelectron tomography / negative staining
AuthorsMeschede IP / Ovenden NC / Seabra MC / Futter CE / Votruba M / Cheetham ME / Burgoyne T
Funding support United Kingdom, 4 items
OrganizationGrant numberCountry
Wellcome Trust093445 United Kingdom
Medical Research Council (MRC, United Kingdom)G0700949 United Kingdom
Wellcome Trust205041 United Kingdom
Medical Research Council (MRC, United Kingdom)G108523 United Kingdom
CitationJournal: Proc Natl Acad Sci U S A / Year: 2020
Title: Symmetric arrangement of mitochondria:plasma membrane contacts between adjacent photoreceptor cells regulated by Opa1.
Authors: Ingrid P Meschede / Nicholas C Ovenden / Miguel C Seabra / Clare E Futter / Marcela Votruba / Michael E Cheetham / Thomas Burgoyne /
Abstract: Mitochondria are known to play an essential role in photoreceptor function and survival that enables normal vision. Within photoreceptors, mitochondria are elongated and extend most of the inner- ...Mitochondria are known to play an essential role in photoreceptor function and survival that enables normal vision. Within photoreceptors, mitochondria are elongated and extend most of the inner-segment length, where they supply energy for protein synthesis and the phototransduction machinery in the outer segment, as well as acting as a calcium store. Here, we examined the arrangement of the mitochondria within the inner segment in detail using three-dimensional (3D) electron microscopy techniques and show they are tethered to the plasma membrane in a highly specialized arrangement. Remarkably, mitochondria and their cristae openings align with those of neighboring inner segments. The pathway by which photoreceptors meet their high energy demands is not fully understood. We propose this to be a mechanism to share metabolites and assist in maintaining homeostasis across the photoreceptor cell layer. In the extracellular space between photoreceptors, Müller glial processes were identified. Due to the often close proximity to the inner-segment mitochondria, they may, too, play a role in the inner-segment mitochondrial arrangement as well as metabolite shuttling. OPA1 is an important factor in mitochondrial homeostasis, including cristae remodeling; therefore, we examined the photoreceptors of a heterozygous knockout mouse model. The cristae structure in the photoreceptors was not greatly affected, but the mitochondria were enlarged and had reduced alignment to neighboring inner-segment mitochondria. This indicates the importance of key regulators in maintaining this specialized photoreceptor mitochondrial arrangement.
History
DepositionJun 7, 2020-
Header (metadata) releaseJul 1, 2020-
Map releaseJul 1, 2020-
UpdateJul 22, 2020-
Current statusJul 22, 2020Processing site: PDBe / Status: Released

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Structure visualization

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  • Imaged by UCSF Chimera
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  • Solid view (volume rendering)
  • Imaged by UCSF Chimera
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Supplemental images

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Map

FileDownload / File: emd_11147.map.gz / Format: CCP4 / Size: 755.8 MB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationHeterozygous Opa1 knockout mouse photoreceptor inner segment mitochondria
Voxel sizeX=Y=Z: 5.867 Å
Density
Minimum - Maximum-128 - 127
Average (Standard dev.)48.076553 (±22.238018)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-60
Dimensions26722672111
Spacing26722672111
CellA: 15676.624 Å / B: 15676.624 Å / C: 651.237 Å
α=β=γ: 90.0 °

CCP4 map header:

modeenvelope stored as signed bytes (from -128 lowest to 127 highest)
Å/pix. X/Y/Z5.8675.8675.867
M x/y/z26722672111
origin x/y/z0.0000.0000.000
length x/y/z15676.62415676.624651.237
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ250250250
MAP C/R/S123
start NC/NR/NS00-60
NC/NR/NS26722672111
D min/max/mean-128.000127.00048.077

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Supplemental data

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Sample components

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Entire : Mouse retina

EntireName: Mouse retina
Components
  • Cell: Mouse retina

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Supramolecule #1: Mouse retina

SupramoleculeName: Mouse retina / type: cell / ID: 1 / Parent: 0 / Details: Mouse photoreceptors
Source (natural)Organism: Mus musculus (house mouse)

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Experimental details

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Structure determination

Methodnegative staining
Processingelectron tomography
Aggregation statetissue

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Sample preparation

BufferpH: 7.4
StainingType: POSITIVE / Material: Lead Citrate and Uranyl Acetate
Sugar embeddingMaterial: Epon
SectioningUltramicrotomy - Instrument: Leica EM UC6 / Ultramicrotomy - Temperature: 300 K / Ultramicrotomy - Final thickness: 150 nm
Fiducial markerManufacturer: BBI Solutions / Diameter: 10 nm

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Electron microscopy

MicroscopeJEOL 1400
Electron beamAcceleration voltage: 120 kV / Electron source: TUNGSTEN HAIRPIN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN ORIUS SC1000 (4k x 2.7k) / Average electron dose: 50.0 e/Å2

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Image processing

Final reconstructionNumber images used: 120

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