TY - JOUR
T1 - Three-dimensional architecture of murine rod cilium revealed by Cryo-EM
AU - Wensel, Theodore G.
AU - Gilliam, Jared C.
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2015
Y1 - 2015
N2 - The connecting cilium of the rod photoreceptor is a tubular structure that bridges two adjacent cellular compartments, the inner segment, the major site of biosynthesis and energy metabolism, and the outer segment, a highly specialized ciliary structure responsible for phototransduction. The connecting cilium allows for active processes of protein sorting and transport to occur between them. Mutations affecting the cargo, their transporters, and the structural components of the primary cilium and basal body lead to aberrant trafficking and photoreceptor cell death. Understanding the overall design of the cilium, its architectural organization, and the function of varied protein complexes within the structural hierarchy of the cilium requires techniques for visualizing their native three-dimensional structures at high magnification. Here we describe methods for isolating retinas from mice, purifying fragments of rod cells that include much of the inner segment and the rod photoreceptor cilia, vitrifying the cell fragments, and determining their structures by cryo-electron tomography.
AB - The connecting cilium of the rod photoreceptor is a tubular structure that bridges two adjacent cellular compartments, the inner segment, the major site of biosynthesis and energy metabolism, and the outer segment, a highly specialized ciliary structure responsible for phototransduction. The connecting cilium allows for active processes of protein sorting and transport to occur between them. Mutations affecting the cargo, their transporters, and the structural components of the primary cilium and basal body lead to aberrant trafficking and photoreceptor cell death. Understanding the overall design of the cilium, its architectural organization, and the function of varied protein complexes within the structural hierarchy of the cilium requires techniques for visualizing their native three-dimensional structures at high magnification. Here we describe methods for isolating retinas from mice, purifying fragments of rod cells that include much of the inner segment and the rod photoreceptor cilia, vitrifying the cell fragments, and determining their structures by cryo-electron tomography.
KW - Cryo-electron tomography
KW - Intraflagellar transport
KW - Primary cilia
KW - Rod outer segment
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U2 - 10.1007/978-1-4939-2330-4_18
DO - 10.1007/978-1-4939-2330-4_18
M3 - Article
AN - SCOPUS:84923675640
SN - 1064-3745
VL - 1271
SP - 267
EP - 292
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -