Isolation of intraflagellar transport trains.
Bottom Line: Moreover, the particles forming isolated IFT trains are structurally similar to the individual particles found in the ∼17S gradient peak.Our results provide the first direct evidence that ∼17S particles do indeed compose the IFT trains.The paper also represents the first isolation of the IFT trains, and opens new possibilities for higher resolution studies on their structure and how particles are attached to each other to form the particle trains.
Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.Show MeSH
Related in: MedlinePlus
Mentions: The gradient pellet was resuspended in buffer, negatively stained, and then analyzed by electron microscopy. It was found to contain roundish particles, frequently arranged into sinuous, flexible strings of variable lengths, which are often associated with what appear to be membrane vesicles (Fig. 4). The overall appearance of this material is quite similar to the IFT immunoreactive beaded structures associated with microtubule doublets detected in whole mount negatively stained axonemes (see Figs. 1 and 2). Particles were found to be arranged into single or double strings (arrows in Fig. 4A) or to form thread-like arrays (arrowheads in Fig. 4A; see also Fig. 4C for a higher magnification image). Frequently, they were associated with membrane patches or vesicles (white arrowheads in Figs. 4B and 4D).
Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.