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Isolation of intraflagellar transport trains.

Mencarelli C, Mitchell A, Leoncini R, Rosenbaum J, Lupetti P - Cytoskeleton (Hoboken) (2013)

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.

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Related in: MedlinePlus

A–B. Low magnification electron microscopy views of the material in the gradient pellet after negative staining, showing the presence of sinuous arrays of particles with different lengths and spatial arrangements. Arrows in A point to single strings of particles, while arrowheads indicate double strings or thread-like arrays of particles. In B the frequent association of particles with membrane vesicles (white arrowheads) is shown. Higher magnification views of two arrays of particles are shown in C–D; white arrowheads in D indicate membrane vesicles.
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fig04: A–B. Low magnification electron microscopy views of the material in the gradient pellet after negative staining, showing the presence of sinuous arrays of particles with different lengths and spatial arrangements. Arrows in A point to single strings of particles, while arrowheads indicate double strings or thread-like arrays of particles. In B the frequent association of particles with membrane vesicles (white arrowheads) is shown. Higher magnification views of two arrays of particles are shown in C–D; white arrowheads in D indicate membrane vesicles.

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).


Isolation of intraflagellar transport trains.

Mencarelli C, Mitchell A, Leoncini R, Rosenbaum J, Lupetti P - Cytoskeleton (Hoboken) (2013)

A–B. Low magnification electron microscopy views of the material in the gradient pellet after negative staining, showing the presence of sinuous arrays of particles with different lengths and spatial arrangements. Arrows in A point to single strings of particles, while arrowheads indicate double strings or thread-like arrays of particles. In B the frequent association of particles with membrane vesicles (white arrowheads) is shown. Higher magnification views of two arrays of particles are shown in C–D; white arrowheads in D indicate membrane vesicles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4060975&req=5

fig04: A–B. Low magnification electron microscopy views of the material in the gradient pellet after negative staining, showing the presence of sinuous arrays of particles with different lengths and spatial arrangements. Arrows in A point to single strings of particles, while arrowheads indicate double strings or thread-like arrays of particles. In B the frequent association of particles with membrane vesicles (white arrowheads) is shown. Higher magnification views of two arrays of particles are shown in C–D; white arrowheads in D indicate membrane vesicles.
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).

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Sciences, University of Siena, Via Aldo Moro 2, 53100, Siena, Italy.

Show MeSH
Related in: MedlinePlus