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Sawai, Satoshi

Kyoto University Research Information Repository

Title High-throughput Analysis of Spatio-temporal multicellulardynamicsAuthor(s)Sawai, SatoshiCitation物性研究 (2007), 87(4): 560-560Issue Date2007-01-20URL http://hdl.handle.net/2433/110745RightType Departmental Bulletin PaperTextversionpublisherKyoto University研究会報告High-throughput Analysis of Spatio圃temporalmulticellular dynamics 津井哲 (SatoshiSawai) JST ERATO複雑系生命プロジヱクトssawai@complex.c.u-tokyo.ac.jp I wil present molecular genetics of a ∞mplex spatio-temporal patterning during the development of social amoeba Dictyostelium. Nutrient-deprived amoebae aggregate to form a multicellular structure by chemotaxis， moving towards propagating waves of cyclic AMP that are relayed from cell to cel¥. Organizing centers are dynamic entities consisting of ∞res of outwardly rotating spiral waves that self-organize in a homogeneous undifferentiated cell population. The number of cores determines the size of aggregation territory and hence the size of mature fruiting-bodies. Here we see a hallmark of self-organization -an emergena of a spatio-temporal pa社ernthrough amplification of initial fluctuations. Using wavelet analysis and a rigorous detection of spatial phase singularities by the time-delay embedding technique， we discovered that mutants of the cyc¥ic AMP/protein kinase A pathway fail to organize coherent long-range wave territories， due to the appearance of numerous spiral cores. A theoretical model that incorporates the current understanding of this pathway suggests that auto・regulation of cell excitability mediated by protein kinase A acts to optimize the number of signaling centers. I wil discuss how the feedback mechanism is able to effectively make use of random firing events to achieve the desired tissue-size. We have also examined over 2，500 mutagenized clones for defects in various steps in this proass， using semi-automated highly parallel time-Iapse imaging and gene sequencing to visualize at millimeter scales al stages of the life cyc¥e with the goal of a complete re∞rd of the temporal and spatial dynamics of each mutant. Inc¥uded in this analysis is the onset and evolution of traveling cAMP waves， characteristic of early development in this organism; the transition from stationary signaling cells to cells streaming toward an organizing center; and the motion of the multicellular slug as itforms a mature fruiting body. 1 have introduced a coarse-grained phenotypic space for c¥ustering mutants in the form of a ‘phenotypic array¥Approximately 40/0 of the clonallines created in an unbiased forward screen were mutant at one of the life-cyc¥e stages. Many of these， along with known mutants， could be ordered by hierarchical clustering into functional groups. Among the mutations identified were independent occurrenωs of known genes and new mutants in common phenotype c¥usters， and mutant phenotypes originating from intergenic insertions. The resulting dataset altows one to search and retrieve life cycle movies and analysis on a gene-by-gene and phenotype-by-phenotype basis. -560-

High-throughput Analysis of Spatio-temporal multicellular dynamics

http://repository.kulib.kyoto-u.ac.jp/dspace/bitstream/2433/110745/1/KJ00004542013.pdf

High-throughput Analysis of Spatio-temporal multicellular dynamics

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