Abstract
Cryo-electron tomography has been a valuable tool in the analysis of 3D structures of cilia at molecular and cellular levels. It opened a way to reconstruct 3D conformations of proteins in cilia at 3-nm resolution, revealed networks of a number of component proteins in cilia, and has even allowed the study of component dynamics. In particular, we have identified the locations and conformations of all the regular inner and outer dyneins, as well as various regulators such as radial spokes. Since the mid 2000s, cryo-electron tomography has provided us with new knowledge, concepts, and questions in the area of cilia research. Now, after nearly 10 years of application of this technique, we are turning a corner and are at the stage to discuss the next steps. We expect further development of this technique for specimen preparation, data acquisition, and analysis. While combining this tool with other methodologies has already made cryo-electron tomography more biologically significant, we need to continue this cooperation using recently developed biotechnology and cell biology approaches.In this review, we will provide an up-to-date overview of the biological insights obtained by cryo-electron tomography and will discuss future possibilities of this technique in the context of cilia research.Electronic supplementary materialThe online version of this article (doi:10.1186/s13630-014-0012-7) contains supplementary material, which is available to authorized users.
Highlights
Cryo-electron tomography has been a valuable tool in the analysis of 3D structures of cilia at molecular and cellular levels
In motile cilia, we can locate the approximate position of target molecules such as microtubule doublets, axonemal dyneins, and radial spokes based on the ninefold symmetry and 96-nm periodicity along each doublet
In addition to nexin/dynein regulatory chain (DRC), which protrudes between inner dynein arm (IDA) and outer dynein arm (ODA) and links all the adjacent microtubule doublet (MTD), we found two linkers, which we named IDL2 and IDL3 (Figure 3C)
Summary
Cryo-electron tomography has been a valuable tool in the analysis of 3D structures of cilia at molecular and cellular levels. Cryo-electron tomography and subtomogram averaging have been successfully revealing structures of the axoneme.
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