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Published online
doi:10.1083/jcb.200905103
The Journal of Cell Biology, Vol. 187, No. 1, 135-148
The Rockefeller University Press, 0021-9525 $30.00
© Pigino et al.
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Article

Electron-tomographic analysis of intraflagellar transport particle trains in situ



Gaia Pigino1, Stefan Geimer2, Salvatore Lanzavecchia3, Eugenio Paccagnini1, Francesca Cantele3, Dennis R. Diener4, Joel L. Rosenbaum4, and Pietro Lupetti1

1 Dipartimento di Biologia Evolutiva, Università di Siena, 53100 Siena, Italy
2 Zellbiologie/Elektronenmikroskopie, Universität Bayreuth, 95440 Bayreuth, Germany
3 Dipartmento di Chimica Strutturale, Università degli Studi di Milano, 20133 Milano, Italy
4 Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520

Correspondence to Pietro Lupetti: lupetti{at}unisi.it

Intraflagellar transport (IFT) is the bidirectional movement of multipolypeptide particles between the ciliary membrane and the axonemal microtubules, and is required for the assembly, maintenance, and sensory function of cilia and flagella. In this paper, we present the first high-resolution ultrastructural analysis of trains of flagellar IFT particles, using transmission electron microscopy and electron-tomographic analysis of sections from flat-embedded Chlamydomonas reinhardtii cells. Using wild-type and mutant cells with defects in IFT, we identified two different types of IFT trains: long, narrow trains responsible for anterograde transport; and short, compact trains underlying retrograde IFT. Both types of trains have characteristic repeats and patterns that vary as one sections longitudinally through the trains of particles. The individual IFT particles are highly complex, bridged to each other and to the outer doublet microtubules, and are closely apposed to the inner surface of the flagellar membrane.

Abbreviations used in this paper: DIC, differential interference contrast; IFT, intraflagellar transport; MT, microtubule; TEM, transmission electron microscopy; WT, wild type.

© 2009 Pigino et al.
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