Dataset Information

Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin.

ABSTRACT: Messenger ribonucleoprotein particles (mRNPs) move randomly within nucleoplasm before they exit from the nucleus. To further understand mRNP trafficking, we have studied the intranuclear movement of a specific mRNP, the BR2 mRNP, in salivary gland cells in Chironomus tentans. Their polytene nuclei harbor giant chromosomes separated by vast regions of nucleoplasm, which allows us to study mRNP mobility without interference of chromatin. The particles were fluorescently labeled with microinjected oligonucleotides (DNA or RNA) complementary to BR2 mRNA or with the RNA-binding protein hrp36, the C. tentans homologue of hnRNP A1. Using high-speed laser microscopy, we followed the intranuclear trajectories of single mRNPs and characterized their motion within the nucleoplasm. The Balbiani ring (BR) mRNPs moved randomly, but unexpectedly, in a discontinuous manner. When mobile, they diffused with a diffusion coefficient corresponding to their size. Between mobile phases, the mRNPs were slowed down 10- to 250-fold but were never completely immobile. Earlier electron microscopy work has indicated that BR particles can attach to thin nonchromatin fibers, which are sometimes connected to discrete fibrogranular clusters. We propose that the observed discontinuous movement reflects transient interactions between freely diffusing BR particles and these submicroscopic structures.

SUBMITTER: Siebrasse JP

PROVIDER: S-EPMC2629337 | biostudies-literature | 2008 Dec

REPOSITORIES: biostudies-literature

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Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin.

Siebrasse Jan Peter JP Veith Roman R Dobay Akos A Leonhardt Heinrich H Daneholt Bertil B Kubitscheck Ulrich U

Proceedings of the National Academy of Sciences of the United States of America 20081212 51

Messenger ribonucleoprotein particles (mRNPs) move randomly within nucleoplasm before they exit from the nucleus. To further understand mRNP trafficking, we have studied the intranuclear movement of a specific mRNP, the BR2 mRNP, in salivary gland cells in Chironomus tentans. Their polytene nuclei harbor giant chromosomes separated by vast regions of nucleoplasm, which allows us to study mRNP mobility without interference of chromatin. The particles were fluorescently labeled with microinjected ...[more]

PMID: 19074261

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Project description:Suspended particles flowing through complex porous spaces exhibit clogging mechanisms determined by factors including their size, deformability, and the geometry of the confinement. This study describes the clogging of rigid particles in a microfluidic device made up of parallel microchannels that taper from the inlet to the outlet, where the constriction width is approximately equal to the particle size. This converging geometry summarizes the dynamics of clogging in flow channels with constrictions that narrow over multiple length scales. Our novel approach allows the investigation of suspension flow dynamics in confined systems where clogs are formed both by sieving and bridging mechanisms simultaneously. Here, flow tests are conducted at constant driving pressures for different particle volume fractions, and a power-law decay which appears to be peculiar to the channels' tapered geometry is observed in all cases. Compared to non-tapered channels, the power-law behavior shows flowrate decay is significantly weaker in tapered channels. This weaker flowrate decay is explained by the formation of discontinuous clogs within each channel. Micrographs of the clogged channels reveal clogs do not grow continuously from their initial positions around the channels' outlet. Rather, new clogs spanning the width of the channel at their points of inception are successively formed as the cake grows toward the inlet area in each microchannel. The results show changes in particle volume fraction at constant driving pressure affect the clogging rate without impacting the underlying dynamics. Unexpectedly, analyses of the particles packing behavior in the microchannels, and post-clogging permeability of the microfluidic devices, reveal the presence of two distinct regimes of driving pressure, though only a small portion of the total device volume and channels surface area are occupied by clogs, regardless of the particle volume fraction. This novel investigation of discontinuous clogging over multiple particle diameters provides unique insights into additional mechanisms to control flow losses in filtration and other confined systems.

Dataset Information

Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin.

Publications

Discontinuous movement of mRNP particles in nucleoplasmic regions devoid of chromatin.

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