Summary of Research Project Results Under the JSPS FY2000
"Research for the future Program"



1.Research Institution Tokyo Institute of Technology
 
2.Research Area Life Sciences
 
3.Research Field Organism Constructing Mechanisms
 
4.Term of Project FY1996〜FY2000
 
5.Project Number 96L00404
 
6.Title of Project Mechanisms of Cell Fate Determination

7.Projetct Leader
Name Institution,Department Title of Position
Hiroki Nishida Tokyo Institute of Technology, Graduate School of Bioscience and Engineering Associate Prof.

8.Core Members

Names Institution,Department Title of Position
Kazuhiro Makabe Kyoto University, Graduate School of Science Assistant Prof.
Takahito Nishikata Konan University, Faculty of Science and Engineering Associate Prof.
Hidetoshi Saiga Tokyo Metropolitan University, Faculty of Science Associate Prof.

9.Cooperating Researchers

Names Institution,Department Title of Position
Katsumi Takamura Fukuyama University, Faculty of Engineering Associate Prof.
Hiroki Takahashi National Institute for Basic Biology Assistant Prof.
William R. Bates Okanagan University College (Canada) Associate Prof.

10.Summary of Research Results

This project is aimed to investigate how cell fates are determined during early embryogenesis. We selected the urochordates ascidian embryo as a model system because of its simplicity, and performed experimental and molecular approaches.
We carried out cDNA project in which 20,000 arrayed clones (corresponds 10,000 clusters) of maternal massage were sequenced in both terminal to generate EST database, and 3,000 clones were in situ hybridized to reveal mRNA localization/expression in eggs and embryos. The data is available through www. Among the posterior-localized clones, we successfully identified macho-1 mRNA as a muscle determinants in ascidian eggs.
We also investigate the mechanisms by which maternal RNAs are localized in embryos. It was found that short sequences in 3'UTR are sufficient for posterior localization in several clones. Several cDNAs of RNA-binding proteins were isolated as candidates that are involved in localization processes.
Invariant cleavage pattern plays an essential roles in precise partitioning of cytoplasmic determinants and localized egg RNAs. We identified the centrosome-attracting body (CAB) that cause asymmetric positioning of nuclei and hence unequal cleavages.
Ascidian embryos also utilize inductive cell interactions for the formation of notochord and mesenchyme. By investigating these inductive processes in detail, we proposed "Directed Signal and Asymmetric Division Model" that is inferred to occur in various kinds of animal embryos that consist of small cell number.
These progresses would bring significant advance in understanding embryogenesis, and would greatly contribute to understand development of various chordates including human.

11.Key Words

(1)embryogenesis、(2)developmental fates、(3)ascidian
(4)cytoplasmic determinants、(5)cDNA project、(6)localized RNA
(7)cleavage pattern、(8)embryonic induction、(9)RNA-binding protein

12.References

[Reference Articles]
Author Title of Article
Nishida, H. Vegetal egg cytoplasm promotes gastrulation and is responsible for specification of vegetal blastomeres in embryos of the ascidian Halocynthia roretzi.
Journal Volume Year Pages Concerned
Development 122 1996 1271-1279

Author Title of Article
Kumano, G., Yokosawa, H., and Nishida, H. Biochemical evidence for membrane-bound endoderm-specific alkaline phosphatase in larvae of the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Eur. J. Biochem. 240 1996 485-489

Author Title of Article
Yamada, A., and Nishida, H. Distribution of cytoplasmic determinants in unfertilized eggs of the ascidian Halocynthia roretzi.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 206 1996 297-304

Author Title of Article
Nishida, H., and Kumano, G. Analysis of temporal expression of the endoderm-specific alkaline phosphatase during development of the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 39 1997 199-203

Author Title of Article
Bates, W. R., and Nishida, H. Developmental roles of nuclear complex factors released during oocyte maturation in the ascidians Halocynthia roretzi and Boltenia villosa.
Journal Volume Year Pages Concerned
Zool. Sci. 15 1998 69-76

Author Title of Article
Kumano, G., and Nishida, H. Maternal and zygotic expression of the endoderm-specific alkaline phosphatas gene in embryos of the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Dev. Biol. 198 1998 245-252

Author Title of Article
Oka, T., Amikura, R., Kobayashi, S., and Nishida, H. Localization of mitochondrial large ribosomal RNA in myoplasm of the early ascidian embryos.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1998 1-8

Author Title of Article
Chiba, S., Satou, Y., Nishikata, T., and Satoh, N. Isolation and characterization of cDNA clones for epidermis-specific, and muscle-specific genes in Ciona savignyi embryos.
Journal Volume Year Pages Concerned
Zool. Sci. 15 1998 239-246

Author Title of Article
Nishida, H., and Sawada, K. macho-1 encodes localized mRNA in ascidian eggs that specifies muscle fate during embryogenesis.
Journal Volume Year Pages Concerned
Nature 409 2001 724-729

Author Title of Article
Nakatani, Y., Yasuo, H., Satoh, N., and Nishida, H. Basic fibroblast growth factor induces notochord formation and the expression of As-T, a brachyury homolog, during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Development 122 1996 2023-2031

Author Title of Article
Kawaminami, S., and Nishida, H. Induction of trunk lateral cells, the blood cell precursors, during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Dev. Biol. 81 1997 14-20

Author Title of Article
Nakatani Y., and Nishida, H. Ras is an essential component for notochord formation during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Mech. Dev. 68 1997 81-89

Author Title of Article
Kim, G. J., and Nishida, H. Monoclonal antibodies against differentiating mesenchyme cells in larvae of the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Zool. Sci. 15 1998 553-559

Author Title of Article
Nakatani, Y., and Nishida, H. Duration of competence and inducing capacity of blastomeres in notochord induction during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1999 449-453

Author Title of Article
Kim, G. J., and Nishida, H. Suppression of muscle fate by cellular interaction is required for mesenchyme formation during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Dev. Biol. 214 1999 9-22

Author Title of Article
Wada, S., Katsuyama, Y., and Saiga, H. Anterior-posterior patterning of the epidermis by inductive influences from the vegetal hemisphere cells in the ascidian embryo.
Journal Volume Year Pages Concerned
Development 126 1999 4955-4963

Author Title of Article
Kim, G. J., Yamada, A., and Nishida, H. An FGF signal from endoderm and localized factors in the posterior-vegetal egg cytoplasm pattern the mesodermal tissues in the ascidian embryo.
Journal Volume Year Pages Concerned
Development 127 2000 2853-2862

Author Title of Article
Kamei, S., Yajima, I., Yamamoto, H., Kobayashi, A., Makabe, K. W., Yamazaki, H., Hayashi, S. I., and Kunisada, T. Characterization of a novel member of the FGFR family, HrFGFR, in Halocynthia roretzi.
Journal Volume Year Pages Concerned
Biochem Biophys Res Commun. 275 2000 503-508

Author Title of Article
Minokawa, T., Yagi, K., Makabe, K. W., and Nishida, H. Binary specification of nerve cord and notochord cell fates in ascidian embryos.
Journal Volume Year Pages Concerned
Development   2001 印刷中

Author Title of Article
Kim, G. J., and Nishida, H. Role of FGF and MEK signaling pathway in the ascidian embryo
Journal Volume Year Pages Concerned
Dev. Growth Differ.   2001 印刷中

Author Title of Article
Darras, S., and Nishida, H. The BMP signaling pathway is required together with the FGF pathway for notochord induction in the ascidian embryo.
Journal Volume Year Pages Concerned
Development   2001 印刷中

Author Title of Article
Darras, S., and Nishida, H. The BMP/CHORDIN antagonism controls sensory pigment cell specification and differentiation in the ascidian embryo
Journal Volume Year Pages Concerned
Dev. Biol.   2001 印刷中

Author Title of Article
Sasakura, Y., Ogasawara, M., and Makabe, K. W. HrWnt-5: a maternally expressed ascidian Wnt gene with posterior localization in early embryos.
Journal Volume Year Pages Concerned
Int. J. Dev. Biol. 42 1998 573-580

Author Title of Article
Sasakura, Y., Ogasawara, M., and Makabe, K. W.. Maternally localized RNA encoding a serine/threonine prote in kinase in the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Mech. Dve. 76 1998 161-163

Author Title of Article
Kobayashi, A., Sasakura, Y., Ogasawara, M., and Makabe K. W. A maternal RNA encoding smad1/5 is segregated to the animal blastomeres during ascidian development.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1999 419-427

Author Title of Article
Sasakura, Y., and Makabe, K. W. Ascidian Wnt-7 gene is expressed exclusively in the tail neural tube of tailbud embryos.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 210 2000 641-643

Author Title of Article
Kawashima, T., Kawashima, S., Kanehisa, M., Nishida, H., and Makabe, K. W. MAGEST: MAboya Gene Expression patterns and Sequence Tags.
Journal Volume Year Pages Concerned
Nuc. Acids Res. 28 2000 133-135

Author Title of Article
Ogasawara, M., Minokawa, T., Sasakura, Y., Nishida, H., and Makabe, K. W. A large-scale whole-mount in situ hybridization system: rapid one-tube preparation of DIG-labeled RNA probes and high throughput hybridization using 96-well silent screen plates
Journal Volume Year Pages Concerned
Zool. Sci. 18 2001 187-193

Author Title of Article
Yagi, K., and Makabe, K. W. Isolation of an early neural maker gene abundantly expressed in the nervous system of the ascidian, Halocynthia roretzi.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 211 2001 49-53

Author Title of Article
Makabe, K. W. et al. Large-scale cDNA analysis of the maternal genetic information in the egg of the ascidian Halocynthia roretzi, for the gene expression catalog during development
Journal Volume Year Pages Concerned
Development   2001 印刷中

Author Title of Article
Hibino, T., Nishikata T., and Nishida, H. Centrosome-attracting body: A novel structure closely related to unequal cleavages in the ascidian embryo.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 40 1998 85-95

Author Title of Article
Nishikata, T., Hibino, T., and Nishida, H. The centrosome-attracting body, microtubule system, and posterior egg cytoplasm are involved in positioning of cleavage planes in the ascidian embryos
Journal Volume Year Pages Concerned
Dev. Biol. 209 1999 72-85

Author Title of Article
Iseto T., and Nishida, H. Ultrastructual studies on the centrosome-attracting body: Specific electron-dense matrix and implications of it's role in unequal cleavage
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1999 601-609

Author Title of Article
Nishikata, T., and Wada, M. Molecular characterization of myoplasmin-Cl: a cytoskeletal component localized in the myoplasm of ascidian eggs.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 206 1996 72-79

Author Title of Article
Wada, M. R., Ohtani Y., Shibata, Y., Tanaka, K. J., Tanimoto, N., and Nishikata, T. An alternatively spliced gene encoding a Y-box protein showing maternal expression and tissue-specific zygotic expression in the ascidian embryos.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 40 1998 631-640

Author Title of Article
Chiba, S., Miki, Y., Ashida, K., Wada, M. R., Tanaka, K. J., Nakamori, R., and Nishikata, T. Interactions between cytoskeletal components during ooplasmic segregation in the myoplasm of ascidian eggs.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1999 265-272

Author Title of Article
Kawashima, T., Murakami, A. R., Ogasawara, M., Tanaka, K. J., Isoda, R., Sasakura, Y., Nishikata, T., Okano, H., and Makabe, K. W. Expression patterns of musashi homologus of the ascidians, Halocynthia roretzi and Ciona intestinalis.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 210 2000 162-165

Author Title of Article
Tanaka, K. J., Kawamura, H., Matsugu, H., and Nishikata, T. An ascidian glycine-rich RNA-binding protein is not induced by temperature-stress but is expressed under a genetic program during embryogenesis.
Journal Volume Year Pages Concerned
Gene 243 2000 207-214

Author Title of Article
Tanaka, K. J., Kawamura, H., and Nishikata, T. The transcript coding for an RNA-binding protein is localized in the anterior side of the ascidian 2-cell stage embryo.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 210 2000 464-466

Author Title of Article
Sasakura, Y., Ogasawara, M., and Makabe, K. W. Two pathways of maternal RNA localization at the posterior-vegetal cytoplasm in early ascidian embryos.
Journal Volume Year Pages Concerned
Dev. Biol. 220 2000 365-378

Author Title of Article
Wada, S., and Saiga, H. Cloning and embryonic expression of Hrsna, a snail family gene of the ascidian Halocynthia roretzi: implication in the origins of mechanisms of mesoderm specification and body axis formation in chordates.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 41 1999 9-18

Author Title of Article
Wada, S., and Saiga, H. Vegetal cell fate specification and anterior neuroectoderm formation by Hroth, the ascidian homologue of orthodenticle/otx.
Journal Volume Year Pages Concerned
Mech. Dev. 82 1999 67-77

Author Title of Article
Katsuyama, Y., Sato, Y., Wada, S. and Saiga, H. Ascidian tail formation requires caudal function
Journal Volume Year Pages Concerned
Dev. Biol. 213 1999 257-268

Author Title of Article
Sasakura, Y., and Makabe, K. W. A gene encoding a new ONECUT class homeodomain protein in the ascidian Halocynthia roretzi functions in the differentiation and specification of neural cells in ascidian embryogenesis.
Journal Volume Year Pages Concerned
Mech. Dev.   2001 印刷中

Author Title of Article
Tanaka, J. K., Chiba, S., and Nishikata, T. Two distinct cell types identified in the asidian notochord
Journal Volume Year Pages Concerned
Zool. Sci. 13 1996 725-730

Author Title of Article
Hori, S., Saitoh, T., Matsumoto, M., Makabe, K. W., and Nishida, H. A Notch homologue from Halocynthia roretzi is preferentially expressed in the central nervous system during ascidian embryogenesis.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 207 1997 371-380

Author Title of Article
Hirano T., and Nishida, H. Developmental fates of larval tissues after metamorphosis in ascidian, Halocynthia roretzi. I. Origin of mesodermal tissues of the juvenile.
Journal Volume Year Pages Concerned
Dev. Biol. 192 1997 199-210

Author Title of Article
Takahashi, H., Ishida, K., Makabe, K. W., and Satoh, N. Isolation of cDNA clones for genes that are expressed in the tail region of the ascidian tailbud embryo.
Journal Volume Year Pages Concerned
Int. J. Dev. Biol. 41 1997 691-698

Author Title of Article
Takamura, K. Nervous network in larvae of the ascidian Ciona intestinalis.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 208 1998 1-8

Author Title of Article
Takamura, K. What are the ascidian test cells? Some suggestions for their origin and functions
Journal Volume Year Pages Concerned
Zygote 6 1998 S75-S76

Author Title of Article
Katsuyama, Y., and Saiga, H. Retinoic acid affects patterning along the anterior-posterior axis of the ascidian embryo.
Journal Volume Year Pages Concerned
Dev. Growth Differ. 40 1998 413-422

Author Title of Article
Yamada, A., and Nishida, H. Distinct parameters are involved in controlling the round of cell division in each tissue during ascidian embryogenesis.
Journal Volume Year Pages Concerned
J. Exp. Zool. 284 1999 379-391

Author Title of Article
Wada, M. R., Murakami, A. R., Kawamura, H., Nakamori, R., Tanaka, K. J., Nakayama, H., and Nishikata, T. The expression of the protochordate homologue of the proteasome regulatory subunit Rpn12 is transcriptionally, and post-translationally regulated during cleavage stage.
Journal Volume Year Pages Concerned
Zool. Sci. 16 1999 125-129

Author Title of Article
Hirano, T., and Nishida, H. Developmental fates of larval tissues after metamorphosis in the ascidian, Halocynthia roretzi. II. Origin of endodermal tissues of the juvenile.
Journal Volume Year Pages Concerned
Dev. Genes Evol. 210 2000 55-63

Author Title of Article
Kobayashi, K., and Nishida, H. Nuclear plasticity and timing mechanisms of the initiation of alkaline phosphatase expression in cytoplasm-transferred blastomeres of ascidians
Journal Volume Year Pages Concerned
Dev. Biol.   2001 印刷中

[Reference Books]
Author Title of Book
Hiroki Nishida et al. (Joint work) Current Developmental Biology
Publisher Year Pages
Kodan-sha 1996 168

Author Title of Book
Hiroki Nishida et al. (Joint work) Sensors in Cognition: Trick of Signaling in Life
Publisher Year Pages
Yoshioka Shoten 1997 127

Author Title of Book
Kazuhiro Makabe Cloning without trouble
Publisher Year Pages
Syujun-sha 1997 208

Author Title of Book
Takahito Nishikata Dealing Protein without Fear
Publisher Year Pages
Syujun-sha 1997 204

Author Title of Book
Nishida H., Makabe K., Saiga H. et al. (Joint work) Biology of Ascidian
Publisher Year Pages
Tokyo University Press 1998 258

Author Title of Book
Hiroki Nishida et al. (Joint work) Revealing Mystery of Animal Body Formation
Publisher Year Pages
Tokyo Kagaku Dojin 2001 印刷中


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