The chromosome peripheral proteins play an active role in chromosome dynamics

Matsunaga, Sachihiro and Fukui, Kiichi. "The chromosome peripheral proteins play an active role in chromosome dynamics" , vol. 1, no. 2, 2010, pp. 157-164. https://doi.org/10.1515/bmc.2010.01

Characterization of somatic embryogenesis initiated from the Arabidopsis shoot apex

Somatic embryogenesis is one of the best examples of the remarkable developmental plasticity of plants, in which committed somatic cells can dedifferentiate and acquire the ability to form an embryo and regenerate an entire plant. In Arabidopsis thaliana, the shoot apices of young seedlings have been reported as an alternative tissue source for somatic embryos (SEs) besides the widely studied zygotic embryos taken from siliques. Although SE induction from shoots demonstrates the plasticity of plants more clearly than the embryo-to-embryo induction system, the underlying developmental and molecular mechanisms involved are unknown. Here we characterized SE formation from shoot apex explants by establishing a system for time-lapse observation of explants during SE induction. We also established a method to distinguish SE-forming and non-SE-forming explants prior to anatomical SE formation, enabling us to identify distinct transcriptome profiles of these two explants at SE initiation. We show that embryonic fate commitment takes place at day 3 of SE induction and the SE arises directly, not through callus formation, from the base of leaf primordia just beside the shoot apical meristem (SAM), where auxin accumulates and shoot-root polarity is formed. The expression domain of a couple of key developmental genes for the SAM transiently expands at this stage. Our data demonstrate that SE-forming and non-SE-forming explants share mostly the same transcripts except for a limited number of embryonic genes and root genes that might trigger the SE-initiation program. Thus, SE-forming explants possess a mixed identity (SAM, root and embryo) at the time of SE specification

In vivo manipulation of fluorescently labeled organelles in living cells by multiphoton excitation

Wataru Watanabe, Sachhiro Matsunaga, Tsunehito Higashi, Kiichi Fukui, and Kazuyoshi Itoh "In vivo manipulation of fluorescently labeled organelles in living cells by multiphoton excitation," Journal of Biomedical Optics 13(3), 031213 (1 May 2008). https://doi.org/10.1117/1.293940

Femtosecond laser manipulation of subcellular organelles in living cells

European Conferences on Biomedical Optics 2005, 2005, Munich, GermanyWataru Watanabe, Tomoko Shimada, Kazuyoshi Itoh, Sachihiro Matsunaga, Kiichi Fukui, "Femtosecond laser manipulation of subcellular organelles in living cells," Proc. SPIE 5863, Therapeutic Laser Applications and Laser-Tissue Interactions II, 58630B (26 August 2005); https://doi.org/10.1117/12.633079

TALE-mediated plant genome visualization

Live imaging of the dynamics of nuclear organization provides the opportunity to uncover the mechanisms responsible for four-dimensional genome architecture. Here, we describe the use of fluorescent protein (FP) fusions of transcription activator-like effectors (TALEs) to visualize endogenous genomic sequences in Arabidopsis thaliana. The ability to engineer sequence-specific TALEs permits the investigation of precise genomic sequences. We could detect TALE-FP signals associated with centromeric, telomeric, and rDNA repeats and the signal distribution was consistent with that observed by fluorescent in situ hybridization. TALE-FPs are advantageous because they permit the observation of intact tissues. We used our TALE-FP method to investigate the nuclei of several multicellular plant tissues including roots, hypocotyls, leaves, and flowers. Because TALE-FPs permit live-cell imaging, we successfully observed the temporal dynamics of centromeres and telomeres in plant organs. Fusing TALEs to multimeric FPs enhanced the signal intensity when observing telomeres. We found that the mobility of telomeres was different in subnuclear regions. Transgenic plants stably expressing TALE-FPs will provide new insights into chromatin organization and dynamics in multicellular organisms

Nucleolin functions in nucleolus formation and chromosome congression

Ma N., Matsunaga S., Takata H., et al. Nucleolin functions in nucleolus formation and chromosome congression. Journal of Cell Science, 120, 12, 2091. https://doi.org/10.1242/jcs.008771

Nanosurgery of sub-cellular organelles in living cells using a femtosecond laser oscillator

Lasers and Applications in Science and Engineering, 2006, San Jose, California, United StatesWataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Hiroshi Ishii, Tsunehito Higashi, Kiichi Fukui, Kazuyoshi Itoh, "Nanosurgery of sub-cellular organelles in living cells using a femtosecond laser oscillator," Proc. SPIE 6108, Commercial and Biomedical Applications of Ultrafast Lasers VI, 610804 (28 February 2006); https://doi.org/10.1117/12.645474

The nuclear scaffold protein SAF-A is required for kinetochore-microtubule attachment and contributes to the targeting of Aurora-A to mitotic spindles

Ma N., Matsunaga S., Morimoto A., et al. The nuclear scaffold protein SAF-A is required for kinetochore-microtubule attachment and contributes to the targeting of Aurora-A to mitotic spindles. Journal of Cell Science, 124, 3, 394. https://doi.org/10.1242/jcs.063347

H1.X with different properties from other linker histones is required for mitotic progression

AbstractWe report here the characterization of H1.X, a human histone H1 subtype. We demonstrate that H1.X accumulates in the nucleolus during interphase and is distributed at the chromosome periphery during mitosis. In addition, the results of fluorescence recovery after photobleaching indicate that the exchange of H1.X on and off chromatin is faster than that of the other H1 subtypes. Furthermore, RNA interference experiments reveal that H1.X is required for chromosome alignment and segregation. Our results suggest that H1.X has important functions in mitotic progression, which are different from those of the other H1 subtypes

Intracellular disruption of mitochondria in a living HeLa cell with a 76-MHz femtosecond laser oscillator

Shimada T., Watanabe W., Matsunaga S., et al. Intracellular disruption of mitochondria in a living HeLa cell with a 76-MHz femtosecond laser oscillator. Optics Express, 13, 24, 9869. https://doi.org/10.1364/OPEX.13.009869