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Title: Defining the layers of a sensory cilium with STORM and cryoelectron nanoscopy

Abstract

Primary cilia carry out numerous signaling and sensory functions, and defects in them, “ciliopathies,” cause a range of symptoms, including blindness. Understanding of their nanometer-scale ciliary substructures and their disruptions in ciliopathies has been hindered by limitations of conventional microscopic techniques. In this work, we have combined cryoelectron tomography, enhanced by subtomogram averaging, with superresolution stochastic optical reconstruction microscopy (STORM) to define subdomains within the light-sensing rod sensory cilium of mouse retinas and reveal previously unknown substructures formed by resident proteins. Domains are demarcated by structural features such as the axoneme and its connections to the ciliary membrane, and are correlated with molecular markers of subcompartments, including the lumen and walls of the axoneme, the membrane glycocalyx, and the intervening cytoplasm. Within this framework, we report spatial distributions of key proteins in wild-type (WT) mice and the effects on them of genetic deficiencies in 3 models of Bardet–Biedl syndrome.

Authors:
 [1];  [1];  [1];  [1]; ORCiD logo [2];  [1]; ORCiD logo [1]
  1. Baylor College of Medicine, Houston, TX (United States)
  2. McGovern Medical School, Houston, TX (United States); Yale School of Medicine, New Haven, CT (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE; National Institutes of Health (NIH)
OSTI Identifier:
1596756
Grant/Contract Number:  
[AC02-76SF00515; R01-EY007981; R01-EY026545; P30-EY002520; F32-EY027171; F31-EY028025; P41-GM103832; Q0035; AU-1714]
Resource Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
[ Journal Volume: 116; Journal Issue: 47]; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; primary cilia; photoreceptors; ciliopathies; superresolution; cryoelectron tomography

Citation Formats

Robichaux, Michael A., Potter, Valencia L., Zhang, Zhixian, He, Feng, Liu, Jun, Schmid, Michael F., and Wensel, Theodore G. Defining the layers of a sensory cilium with STORM and cryoelectron nanoscopy. United States: N. p., 2019. Web. doi:10.1073/pnas.1902003116.
Robichaux, Michael A., Potter, Valencia L., Zhang, Zhixian, He, Feng, Liu, Jun, Schmid, Michael F., & Wensel, Theodore G. Defining the layers of a sensory cilium with STORM and cryoelectron nanoscopy. United States. doi:10.1073/pnas.1902003116.
Robichaux, Michael A., Potter, Valencia L., Zhang, Zhixian, He, Feng, Liu, Jun, Schmid, Michael F., and Wensel, Theodore G. Tue . "Defining the layers of a sensory cilium with STORM and cryoelectron nanoscopy". United States. doi:10.1073/pnas.1902003116.
@article{osti_1596756,
title = {Defining the layers of a sensory cilium with STORM and cryoelectron nanoscopy},
author = {Robichaux, Michael A. and Potter, Valencia L. and Zhang, Zhixian and He, Feng and Liu, Jun and Schmid, Michael F. and Wensel, Theodore G.},
abstractNote = {Primary cilia carry out numerous signaling and sensory functions, and defects in them, “ciliopathies,” cause a range of symptoms, including blindness. Understanding of their nanometer-scale ciliary substructures and their disruptions in ciliopathies has been hindered by limitations of conventional microscopic techniques. In this work, we have combined cryoelectron tomography, enhanced by subtomogram averaging, with superresolution stochastic optical reconstruction microscopy (STORM) to define subdomains within the light-sensing rod sensory cilium of mouse retinas and reveal previously unknown substructures formed by resident proteins. Domains are demarcated by structural features such as the axoneme and its connections to the ciliary membrane, and are correlated with molecular markers of subcompartments, including the lumen and walls of the axoneme, the membrane glycocalyx, and the intervening cytoplasm. Within this framework, we report spatial distributions of key proteins in wild-type (WT) mice and the effects on them of genetic deficiencies in 3 models of Bardet–Biedl syndrome.},
doi = {10.1073/pnas.1902003116},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = [47],
volume = [116],
place = {United States},
year = {2019},
month = {11}
}

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