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- Ebo: Experimental Biology Online Annual 1998.
Details if other :. Thanks for telling us about the problem. Return to Book Page. Preview — Ebo by Christopher R. Bridges Editor ,. Dale Sanders Editor. Adam Curtis Editor. Though it is a pleasure to write a short foreword to this collection of excellent scientific papers covering a range of biological topics, the rather depressing feature is the small number of papers. All-electronic publishing is developing and your Editors do have great faith in it. One problem for potential authors has been the reluctance of the abstracting journals to pa Though it is a pleasure to write a short foreword to this collection of excellent scientific papers covering a range of biological topics, the rather depressing feature is the small number of papers.
One problem for potential authors has been the reluctance of the abstracting journals to pay any attention to electronic journals - perhaps Springer should make a rapid move in this area and start the first all-electronic journal abstracting this type of literature. However, even the paper citation journals are starting to pay attention to the medium. The particular advantages of all-electronic publishing are beginning to emerge more clearly and it is clear that publishing video material is a unique advantage of our format.
Several papers took advantage of this - for example those by Riehle and others on cell behaviour in tunnels, by Bereiter-Hahn and Voss on zonation in the plasmalemma and by Pavlikova, Zicha, Chaloupkova and Vesely on cell motility of tumour cells. These papers made essential and extensive use of video material, publishing some material of great originality. The work on cell pola rity and calcium ions in Fucus embryos by Brownlee, Manison and Anning used animation to present their results in an especially clear way.
The facility of use of animation is another special advantage of our type of publication that should be more widely used. Get A Copy. Paperback , pages. More Details Most of nucleoids remained apparently at the same positions, showing short-distance random motions apparently confined to mitochondrial network, as one can see from overlay of tracks images on mitochondria images Fig 9 ; however, no firm conclusion can be made about the confinement of the tracks within mitochondria, since they were not continuously imaged during tracking time series.
Track mean speed was 0. Only few fast directional displacements per field of view occurred during the observation time window S1 Video. Representative images of a field of view of DMSO-treated cells. Top: two-color SIM image taken before acquisition of the time lapse series. Then we aimed to demonstrate that live cell SIM of SYBR Gold-labelled mitochondrial nucleoids can provide information on a biologically relevant effect.
Mitochondria are linked to microtubules e. Thus, here we tested how motions of mitochondrial nucleoids are affected by microtubule depolymerizing drug nocodazole. S2 Video , detected the tracks of SYBR Gold-stained mitochondrial nucleoids and extracted quantitative track parameters. Results are summarized in Table 2. Interestingly, variation of Instant speed within a track increased from 0.
Thus we speculate that nucleoids are indirectly anchored to microtubule network as it was suggested earlier [ 45 ] , and they become more mobile when the network disappears.
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Obviously, detailed characterization of a putative involvement of microtubules in mitochondrial nucleoids positioning requires further extensive studies, far beyond the scope of the present report. Control of microtubules depolymerization upon nocodazole treatment.
Mitochondrial nucleoids tracks parameters in the absence blue bars and in the presence orange bars of nocodazole. Histograms show distribution of track mean speed B , track maximal speed C and track length D. Particle tracking is described in Materials and Methods section. Direct labelling of DNA with nucleic acid-binding dyes have several advantages over other labelling strategies. Here, we used this strategy. Tracking of mitochondrial nucleoids at resolution below the diffraction limit and with a modest illumination dose has been achieved for the first time thanks to SYBR Gold staining in combination with SIM, which has been chosen since other major super-resolution techniques, STED and SMLM, require much higher illumination doses, which may cause severe phototoxic effects, especially during time lapse imaging.
We observed that at low concentrations SYBR Gold preferentially stained mitochondrial nucleoids, while at high concentrations both chromatin and mtDNA were stained. Unfortunately, mitochondrial nucleoids staining with SYBR Gold is not compatible with conventional IF protocols because it does not resist permeabilization step.
Our study of SYBR Gold emphasizes that other organic dyes widely used in vitro may be useful for fluorescence microscopy or other cell-based technique like flow cytometry. Detected mitochondrial nucleoids are marked as white balls. Mitochondrial nucleoids tracking and visualization by Imaris 8. HeLa cells were incubated for 30 min with mixture of 0.
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LSM microscope, 63x 1. Z-stacks were acquired at each time point; maximum intensity projections are shown. Representative fields of view showing the regions of interest where SYBR Gold fluorescence was measured colored rectangles. A square region is shown marked with white line with higher magnification in the right column. A representative field of view is shown. Deconvolution of datasets was performed.
Cyan squares on the left panels mark the region of interest which is shown at a higher magnification in the right panels. A representative 1. Maximum intensity projection of a z-stack is shown. SIM settings are described in Materials and Methods section. Briefly, frame time 1. Confocal imaging was performed under two settings: 1 blue, the same illumination power as for SIM Intensity profiles across nucleoids on confocal images acquired with Browse Subject Areas?
Click through the PLOS taxonomy to find articles in your field. Abstract Mitochondrial DNA molecules coated with proteins form compact particles called mitochondrial nucleoids. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript Competing interests: The authors have declared that no competing interests exist. Introduction Genetic material of mitochondria consists of a circular Download: PPT. Fig 2. Fig 3. Fig 6. Table 1. Fig 7. Fig 8.
Fig Effect of microtubule depolymerization on the nucleoids motions. Table 2. Effect of microtubule depolymerization on nucleoids motions parameters. Supporting information.
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S1 Video. Effect of microtubules depolymerization on mitochondrial nucleoids motions. S2 Video. S1 Fig. SYBR Gold localization in live cells upon labelling at different concentrations. S2 Fig. S3 Fig.
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S4 Fig. S5 Fig.
S6 Fig. Comparison of acquisition photobleaching in confocal and SIM modes. References 1. Hum Mol Genet. Mechanisms controlling mitochondrial biogenesis and respiration through the thermogenic coactivator PGC Bogenhagen DF.
Mitochondrial DNA nucleoid structure. Biochim Biophys Acta.
Mol Biol Cell. Nature communications. Nucleic Acids Res.