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Published online May 29, 2007
doi:10.1083/jcb.200612010
The Journal of Cell Biology, Vol. 177, No. 5, 917-925
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Nickell et al.
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Article

 Three-dimensional architecture of murine rod outer segments determined by cryoelectron tomography

Stephan Nickell1, Paul S.-H. Park2, Wolfgang Baumeister1, and Krzysztof Palczewski2

1 Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, D-82152 Martinsried, Germany
2 Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106

Correspondence to Wolfgang Baumeister: baumeist{at}biochem.mpg.de

The rod outer segment (ROS) of photoreceptor cells houses all components necessary for phototransduction, a set of biochemical reactions that amplify and propagate a light signal. Theoretical approaches to quantify this process require precise information about the physical boundaries of the ROS. Dimensions of internal structures within the ROS of mammalian species have yet to be determined with the precision required for quantitative considerations. Cryoelectron tomography was utilized to obtain reliable three-dimensional morphological information about this important structure from murine retina. Vitrification of samples permitted imaging of the ROS in a minimally perturbed manner and the preservation of substructures. Tomograms revealed the characteristic highly organized arrangement of disc membranes stacked on top of one another with a surrounding plasma membrane. Distances among the various membrane components of the ROS were measured to define the space available for phototransduction to occur. Reconstruction of segments of the ROS from single-axis tilt series images provided a glimpse into the three-dimensional architecture of this highly differentiated neuron. The reconstructions revealed spacers that likely maintain the proper distance between adjacent discs and between discs and the plasma membrane. Spacers were found distributed throughout the discs, including regions that are distant from the rim region of discs.

S. Nickell and P.S.-H. Park contributed equally to this paper.

Abbreviations used in this paper: AFM, atomic force microscopy; ET, electron tomography; GARP, glutamic acid–rich protein; ROS; rod outer segment; TEM, transmission EM.


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