Purification and Localization of Intraflagellar Transport Particles and Polypeptides
The growth and maintenance of almost all cilia and flagella are dependent on the proper functioning of the process of intraflagellar transport (IFT). This includes the primary cilia of most cells in humans that are in interphase or the Go phase of the cell cycle. The model system for the study of IFT is the flagella of the bi-flagellate green alga Chlamydomonas. It is in this organism that IFT was first discovered, and genetic data from a Chlamydomonas mutant first linked the process of IFT to polycystic kidney disease in humans. The information given in this chapter addresses procedures to purify IFT particles from flagella and localize these particles, and their associated motor proteins, in flagella using light and electron microscopic approaches.
Key words:Flagella Cilia Intraflagellar transport Motility Immunofluorescence Immunogold EM
- 8.D. Signor, K.P. Wedaman, J.T. Orozco, N.D. Dwyer, C.I. Bargmann, L.S. Rose, and J.M. Scholey (1999). Role of a class DHC1b dynein in retrograde transport of IFT motors and IFT raft particles along cilia, but not dendrites, in chemosensory neurons of living Caenorhabditis elegans. J Cell Biol 147 519–530CrossRefPubMedGoogle Scholar
- 10.D.G. Cole, D.R. Diener, A.L. Himelblau, P.L. Beech, J.C. Fuster, and J.L. Rosenbaum (1998). Chlamydomonas kinesin-II-dependent intraflagellar transport (IFT): IFT particles contain proteins required for ciliary assembly in Caenorhabditis elegans sensory neurons. J Cell Biol 141 993–1008CrossRefPubMedGoogle Scholar
- 17.S.H. Hutner, L. Provasoli, Albert Schatz, and C. P. Haskins (1950). Some approaches to the study of the role of metals in the metabolism of microorganisms. Proc Am Philos Soc 94 152–170Google Scholar