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- EMDB-20547: Outer Hair Cell, Lateral Wall -

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Basic information

Entry
Database: EMDB / ID: EMD-20547
TitleOuter Hair Cell, Lateral Wall
Map data
Sample
  • Tissue: Cochlea
Biological speciesCavia porcellus (domestic guinea pig)
Methodelectron tomography / cryo EM / negative staining
AuthorsAuer M
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute on Deafness and Other Communication Disorders United States
CitationJournal: Front Cell Neurosci / Year: 2019
Title: 3D Ultrastructure of the Cochlear Outer Hair Cell Lateral Wall Revealed By Electron Tomography.
Authors: William Jeffrey Triffo / Hildur Palsdottir / Junha Song / David Gene Morgan / Kent L McDonald / Manfred Auer / Robert M Raphael /
Abstract: Outer Hair Cells (OHCs) in the mammalian cochlea display a unique type of voltage-induced mechanical movement termed electromotility, which amplifies auditory signals and contributes to the ...Outer Hair Cells (OHCs) in the mammalian cochlea display a unique type of voltage-induced mechanical movement termed electromotility, which amplifies auditory signals and contributes to the sensitivity and frequency selectivity of mammalian hearing. Electromotility occurs in the OHC lateral wall, but it is not fully understood how the supramolecular architecture of the lateral wall enables this unique form of cellular motility. Employing electron tomography of high-pressure frozen and freeze-substituted OHCs, we visualized the 3D structure and organization of the membrane and cytoskeletal components of the OHC lateral wall. The subsurface cisterna (SSC) is a highly prominent feature, and we report that the SSC membranes and lumen possess hexagonally ordered arrays of particles. We also find the SSC is tightly connected to adjacent actin filaments by short filamentous protein connections. Pillar proteins that join the plasma membrane to the cytoskeleton appear as variable structures considerably thinner than actin filaments and significantly more flexible than actin-SSC links. The structurally rich organization and rigidity of the SSC coupled with apparently weaker mechanical connections between the plasma membrane (PM) and cytoskeleton reveal that the membrane-cytoskeletal architecture of the OHC lateral wall is more complex than previously appreciated. These observations are important for our understanding of OHC mechanics and need to be considered in computational models of OHC electromotility that incorporate subcellular features.
History
DepositionAug 1, 2019-
Header (metadata) releaseFeb 5, 2020-
Map releaseFeb 5, 2020-
UpdateFeb 5, 2020-
Current statusFeb 5, 2020Processing site: RCSB / Status: Released

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Structure visualization

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

emd_20547.png

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Map

FileDownload / File: emd_20547.map.gz / Format: CCP4 / Size: 450 MB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
Voxel sizeX=Y=Z: 8.14 Å
Density
Minimum - Maximum0 - 3906
Average (Standard dev.)1318.3076 (±87.30751)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-225
Dimensions10241024225
Spacing10241024225
CellA: 8335.36 Å / B: 8335.36 Å / C: 1831.5001 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Integer*27
Å/pix. X/Y/Z8.148.148.14
M x/y/z10241024225
origin x/y/z0.0000.0000.000
length x/y/z8335.3608335.3601831.500
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS00-225
NC/NR/NS10241024225
D min/max/mean0.0003906.0001318.308

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Supplemental data

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Sample components

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Entire : Cochlea

EntireName: Cochlea
Components
  • Tissue: Cochlea

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Supramolecule #1: Cochlea

SupramoleculeName: Cochlea / type: tissue / ID: 1 / Parent: 0
Source (natural)Organism: Cavia porcellus (domestic guinea pig)

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Experimental details

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Structure determination

Methodnegative staining, cryo EM
Processingelectron tomography
Aggregation statetissue

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Sample preparation

BufferpH: 7
StainingType: POSITIVE / Material: Uranyl Acetate
Sugar embeddingMaterial: Epon
GridDetails: unspecified
VitrificationCryogen name: NITROGEN
High pressure freezingInstrument: OTHER
Details: The value given for _emd_high_pressure_freezing.instrument is BALTEC. This is not in a list of allowed values set(['LEICA EM PACT2', 'LEICA EM PACT', 'EMS-002 RAPID IMMERSION FREEZER', ...Details: The value given for _emd_high_pressure_freezing.instrument is BALTEC. This is not in a list of allowed values set(['LEICA EM PACT2', 'LEICA EM PACT', 'EMS-002 RAPID IMMERSION FREEZER', 'OTHER', 'LEICA EM HPM100', 'BAL-TEC HPM 010']) so OTHER is written into the XML file.
SectioningUltramicrotomy - Instrument: Leica EM UC6 / Ultramicrotomy - Temperature: 298 K / Ultramicrotomy - Final thickness: 100 nm
Fiducial markerManufacturer: Electron Microscopy Sciences / Diameter: 5 nm

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Electron microscopy

MicroscopeFEI/PHILIPS CM200FEG
Image recordingFilm or detector model: TVIPS TEMCAM-F216 (2k x 2k) / Average electron dose: 10.0 e/Å2
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD

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Image processing

Final reconstructionNumber images used: 130

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