FY2018 Annual Report

 

Abstract

Our team is dedicated to
      (1) methodology development for single-molecule imaging and manipulation at nanometer precisions in living cells, with a special attention paid to high time resolutions (ultrafast single fluorescent-molecule tracking), as well as
     (2) revealing meso~nano-scale processes in signal transduction in/on the cell membrane and the formation and remodeling of the neuronal network, by using the developed single-molecule techniques.
     The smooth liaison between physics/engineering and biomedicine is a key for our research. Based on unique insights we develop by applying single-molecule tracking methods to nano~meso-scale processes occurring in signal transduction and neuronal network formation/modulation, we intend to develop new types of systems molecular biology.
     In the same context, we are now revealing the mechanisms by which the metastable molecular complexes and meso-scale membrane domains, including membrane compartments, raft domains, and protein oligomers, form and work in concert to enable various plasma membrane functions.

1．Staff  

  • Dr. Amine Betul Nuriseria Aladag, Post Doctoral Scholar
  • Dr. An-An Liu, JSPS Research Fellow 
  • Dr. Taka-Aki Tsunoyama, Post Doctoral Scholar
  • Dr. Peng Zhou, Post Doctoral Scholar
  • Dr. Irina Meshcheryakova, Technician
  • Ms. Limin Chen, Technician
  • Ms. Aya Nakamura, Technician
  • Ms. Yuri Nemoto, Technician
  • Ms. Hisae Tsuboi, Technician
  • Mr. Alexey Yudin, Technician
  • Mr. Yoshifumi Maeda, Research Assistant (Part-time)
  • Mr. Shogo Miyagi, Research Assistant (Part-time)
  • Mr. Masaya Negawa, Research Assistant (Part-time)
  • Mr. Takaya Shimabukuro, Research Assistant (Part-time)
  • Ms. Miwako Tokuda, Research Unit Administrator
  • Dr. Akihiro Kusumi, Professor

2．Collaborations

  2.1 Revealing metastable signaling molecular complexes by single-molecule imaging

 - Description: Developing ultrafast 3D single-molecule imaging, and applying it to the dynamics and formation mechanism of the signaling complex in synaptic signaling, Fcepsilon signaling, focal adhesion architecture and signaling, and GPI-anchored proteins’ raft-based signaling

 - Type of collaboration: Joint research

 - Researchers:
      Dr. Takahiro Fujiwara, Associate Professor, Institute of Advanced Studies, Kyoto University
      Dr. Nao Hiramoto-Yamaki, Postdoctoral Research Associate, Institute of Advanced Studies, Kyoto University
      Dr. Kenichi Suzuki, Professor, G-CHAIN, Gifu University

2.2 Elucidation of dynamics and formation mechanisms of cellular signaling complexes by developing new single particle tracking methods

 - Description: Developing fluorescent probes for their applications to single-molecule imaging in living cells, and by using the developed probes, elucidating dynamics and formation mechanisms of cellular signaling complexes induced by various intercellular signaling molecules and alien antigens, including (non-pathogenic) viruses

 - Type of collaboration: Joint research

 - Researchers:
      Dr. Dai-Wen Pang, Professor
      Mr. Bo Tang, PhD candidate
      Ms. Meng-ni Bao, PhD candidate
      Ms. Dan-dan Fu, PhD candidate
      Ms. Jing Li, PhD candidate
      College of Chemistry and Molecular Sciences, Wuhan University, P. R. China          

2.3 Unraveling single-molecule dynamics of neurotransmitter receptors in neurons

 - Description: Examining dynamic monomer-oligomer equilibrium of neurotransmitter receptors and their entrance-exiting dynamics into and out of the synapses

 - Type of collaboration: Joint research

 - Researchers:
      Dr. Jyoji Morise, Assistant Professor, Graduate School of Medicine, Kyoto University
      Dr. Shogo Oka, Professor, Graduate School of Medicine, Kyoto University
      Dr. Kenichi Suzuki, Professor, G-CHAIN, Gifu University
      Dr. Takahiro Fujiwara, Associate Professor, Institute of Advanced Studies, Kyoto University

2.4 Revealing regulation systems of synapse dynamics based on nanoscale morphological analysis

 - Description: Revealing dynamic regulation systems for molecular compositions in the synapse using 3D reconstruction electron microscopy and super-resolution fluorescence microscopy

 - Type of collaboration: Joint research

 - Researchers:
      Dr. Shigeo Okabe, Professor, Graduate School of Medicine, The University of Tokyo
      Dr. Yasunori Inoue, Professor, Graduate School of Engineering, Kyoto University

 

3. Activities and Findings

3.1 Revealing metastable signaling molecular complexes by developing new single-molecule imaging methods

In this project, we develop new single fluorescent-molecule imaging-tracking methods and new fluorescent probes, including ultrafast (world’s fastest) single-molecule imaging, methods for suppressing photobleaching and photoblinking for very long single molecule tracking (several minutes) in living cells, and new fluorescent lipid probes that behave very much like their parent endogenous lipid molecules. By applying the developed methods and probes, we try to reveal the dynamics and formation mechanism of metastable signaling molecular complexes in the context of synaptic signaling, Fcepsilon signaling, focal adhesion architecture and signaling, and GPI-anchored proteins’ raft-based signaling. Two types of major developments on this front in the FY2018 are described below.

Development of super-long single-molecule tracking revealed dynamic-anchorage-induced integrin function

Single-fluorescent-molecule imaging tracking (SMT) is becoming an important tool to study living cells. However, photobleaching and photoblinking (hereafter referred to as photobleaching/photoblinking) of the probe molecules strongly hamper SMT studies of living cells, making it difficult to observe in vivo molecular events and to evaluate their lifetimes (e.g., off rates). The methods used to suppress photobleaching/photoblinking in vitro are difficult to apply to living cells because of their toxicities.
     Here, using 13 organic fluorophores we found that, by combining low concentrations of dissolved oxygen with a reducing-plusoxidizing system, photobleaching/photoblinking could be strongly suppressed with only minor effects on cells, which enabled SMT for as long as 12,000 frames (~7!min at video rate, as compared to the general 10-s-order durations) with ~22-nm single-molecule localization precisions. SMT of integrins revealed that they underwent temporary (< 80-s) immobilizations within the focal adhesion region, which were responsible for the mechanical linkage of the actin cytoskeleton to the extracellular matrix.
 

The class-A GPCR dopamine D2 receptor forms transient dimers stabilized by agonists as revealed by single-molecule tracking

Whether class-A G-protein coupled receptors (GPCRs) exist and work as monomers or dimers has drawn extensive attention. A class-A GPCR dopamine D2 receptor (D2R) is involved in many physiological and pathological processes and diseases, indicating its critical role in proper functioning of neuronal circuits. In particular, D2R homodimers might play key roles in schizophrenia development and amphetamine-induced psychosis.
     Here, using single-molecule imaging, we directly tracked single D2R molecules in the plasma membrane at a physiological temperature of 37˚C, and unequivocally determined that D2R forms transient dimers with a lifetime of 68 ms in its resting state. Agonist addition prolonged the dimer lifetime by a factor of ~1.5, suggesting the possibility that transient dimers might be involved in signaling.

 

3.2 Unraveling the regulation mechanisms for the synaptic structural plasticity by observing dynamics, assembly, and function of neuronal receptors using super-resolution single-molecule imaging and tracking

In this project, we try to understand the mechanisms by which structural synaptic plasticity is induced. To accomplish this goal, we examine, at the level of single molecules, molecular interactions and dynamics in hippocampal neurons in culture. In particular, we examine the dynamic equilibrium of monomers, dimers, oligomers, and clusters of neurotramsmitter receptors and other neuronal molecules, the molecules’ cooperative interactions, including the possibility of phase separation, as well as their entrance-exiting dynamics into and out of the synapses.

 

3.3 Examining and refining our working hypothesis, in which the bulk plasma membrane could be largely considered to be hierarchically organized in three-tiered meso-scale domains, particularly in the context of signal transduction

As described in the home page of our web site, we think the concept of three-tiered meso-scale domain architecture of the plasma membrane provides an excellent perspective on the mechanisms for various functions of the plasma membrane. “Meso” means “between” and the meso-scale generally speaks to the scale between nanometer and micrometer. Often, the actual scale of meso is between 3 and 300 nm. It is an interesting scale where non-living molecules turn into living cells.

The first and most basic tier in this hierarchical architecture is the membrane compartments, formed due to the partitioning of the entire plasma membrane by the actin-based membrane skeleton. We think it is the most basic tier because it is everywhere throughout the plasma membrane and it dominates the movements of all the molecules associated with the plasma membrane.

The second tier is the raft domains, which are localized within the membrane compartments.

The third tier is dynamic protein complexes, with lifetimes often of the order of 0.1 seconds. And so, these are metastable or transient molecular complexes.

Of course, in the real plasma membrane, these three domains coexist in a single membrane and work in concert to enable various plasma membrane functions.

We at the Kusumi lab are examining and refining this working hypothesis. By performing such research, we hope to obtain better perspectives on how the plasma membrane is organized or poised to perform various plasma membrane functions, particularly the signal transduction. Major advancements on this front in the FY2018 are described below.

Revealing the raft domain organization in the plasma membrane by single-molecule imaging of fluorescent ganglioside analogs

Gangliosides have been implicated in a variety of physiological processes, particularly in the formation and function of raft domains in the plasma membrane. However, the scarcity of suitable fluorescent ganglioside analogs had long prevented us from determining exactly how gangliosides perform their functions in the live-cell plasma membrane. With the development of new fluorescent ganglioside analogs made by us and our collaborators, this barrier has been broken. We can now address the dynamic behaviors of gangliosides in the live-cell plasma membrane, using fluorescence microscopy, particularly by single-fluorescent molecule imaging and tracking.
     Single-molecule tracking of fluorescent GM1 and GM3 revealed that these molecules are transiently and dynamically recruited to monomers (monomer-associated rafts) and homodimer rafts of the raftophilic GPI-anchored protein CD59 in quiescent cells, with exponential residency times of 12 and 40 ms, respectively, in a manner dependent on raft–lipid interactions. Upon CD59 stimulation, which induces CD59-cluster signaling rafts, the fluorescent GM1 and GM3 analogs were recruited to the signaling rafts, with a lifetime of 48 ms.
     These results represent the first direct evidence that GPI-anchored receptors and gangliosides interact in a cholesterol-dependent manner. Furthermore, they unequivocally show that gangliosides continually move in and out of rafts that contain CD59 in an extremely dynamic manner, with much higher frequency than expected previously. Such studies would not have been possible without fluorescent ganglioside probes behaving very much like the native parent molecules in various biochemical and conventional microscopy assays.

 

4. Publications

4.1 Journals

ORIGINAL ARTICLES

R. S. Kasai, S. V. Ito, R. M. Awane, T. K. Fujiwara, and A. Kusumi. The class-A GPCR dopamine D2 receptor forms transient dimers stabilized by agonists: detection by single-molecule tracking. Cell Biochem. Biophys. 76, 29-37 (2018). doi: 10.1007/s12013-017-0829-y.

T. A. Tsunoyama, Y. Watanabe, J. Goto, K. Naito, K. G. N. Suzuki, T. K. Fujiwara, and A. Kusumi. Super-long single-molecule tracking reveals dynamic-anchorage-induced integrin function. Nat. Chem. Biol. 14, 497-506 (2018). doi: 10.1038/s41589-018-0032-5.

S. S. Tiwari, Y. M. Shirai, Y. L. Nemoto, K. Kojima, and K. G. N. Suzuki. Native prion protein homodimers are destabilized by oligomeric amyloid β 1-42 species as shown by single-molecule imaging. Neuroreport 29,106-111. (2018). doi:10.1097/WNR.0000000000000916

INVITED REVIEW ARTICLES

K. G. N. Suzuki, H. Ando, N. Komura, M. Konishi, A. Imamura, H. Ishida, M. Kiso, T. K. Fujiwara, and A. Kusumi. Revealing the raft domain organization in the plasma membrane by single-molecule imaging of fluorescent ganglioside analogs. Methods Enzymol. 598, 267-282 (2018). doi: 10.1016/bs.mie.2017.06.038.

4.2 Books and other one-time publications

   Nothing to report

4.3 Oral and Poster Presentations

INVITED PRESENTATIONS

A. Kusumi. Signal transduction by metastable molecular complexes: findings by single-molecule tracking. Biomedicum Helsinki Research Center/University of Helsinki. Biomedicum Helsinki Seminar Series 2018. Helsinki, Finland. October 1, 2018.

A. Kusumi. Signal transduction enabled by very transient molecular complexes: findings by single-molecule tracking. Keynote Lecture. AsiaNano2018 (Asian Conference on Nanoscience and Nanotechnology). Qingdao (Tsintao), China. October 19, 2018.

A. Kusumi. Single-molecule view of the plasma membrane organization that is responsible for signal transduction. The 25th East Asia Joint Symposium. Chongqing, China. October 26, 2018.

A. Kusumi. Signal transduction enabled by very transient molecular complexes: findings by single-molecule tracking. Joint Symposium of ResonanceBio/Global Bio-Imaging (Euro-Bioimaging)/ABiS (Advanced Bioimaging Support). Okinawa Institute of Science and Technology Graduate University (OIST), Okinawa. October 31, 2018.

A. Kusumi. Single-molecule tracking reveals signal transduction by transient molecular complexes. City of Hope Comprehensive Cancer Center. Department of Molecular Medicine Seminar and Special Topics Lecture in Biochemistry and Structural Biology. Los Angels, California, U.S.A. December 13, 2018.

A. Kusumi. Signal transduction by metastable molecular complexes: findings by single-molecule tracking. The 63rd Annual Meeting of the Biophysical Society; Subgroup Symposium “Membrane Structure and Function”. Baltimore, U.S.A. March 2, 2019.

T. A. Tsunoyama. Elongated single-molecule tracking revealed dynamic architecture of focal adhesion. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

Y. L. Nemoto, Fast and slow turnover of AMPA receptor and stargazin in the spine in the time scale of to several 100 s; unraveling by single molecule imaging. The 16th International Membrane Research Forum. Okinawa, Japan. March 19, 2019.

 

ORAL PRESENTATIONS (Selected for Oral Presentation)

R. S. Kasai, T. K. Fujiwara, and A. Kusumi. Transient dimers of GPCRs are responsible for triggering GPCRs’ basic constitutive signals: A finding by the two-color single fluorescent-molecule tracking in living cells. Cold Spring Harbor Laboratory (CSHL) meeting on “Single Biomolecules”. Cold Spring Harbor, New York, U.S.A. August 29, 2018.

T. A. Tsunoyama, K. G. N. Suzuki, T. K. Fujiwara, and A. Kusumi. Super-long single fluorescent-molecule tracking reveals that cell adhesion is induced by dynamic and tension-dependent anchorage of integrin. Cold Spring Harbor Laboratory (CSHL) meeting on “Single Biomolecules”. Cold Spring Harbor, New York, U.S.A. August 30, 2018.

T. K. Fujiwara, and A. Kusumi. Ultrafast single fluorescent-molecule imaging reveals hop diffusion within focal adhesion: Actin-induced compartmentalization of the channels between the focal-adhesion-protein islands. Cold Spring Harbor Laboratory (CSHL) meeting on “Single Biomolecules”. Cold Spring Harbor, New York, U.S.A. August 30, 2018.

K. M. Hirosawa, N. Hiramoto-Yamaki, K. J. Yoshida, S. Nozaki, T. A. Tsunoyama, B. Tang, K. G. N. Suzuki, K. Nakayama, T. K. Fujiwara, and A. Kusumi. LAT vesicles work as a signal transduction platform in immune cells; unraveling by single-molecule imaging. The 63rd Annual Meeting of Biophysical Society (BPS19). Baltimore, Maryland, U.S.A. March 4, 2019.

Y. L. Nemoto, K. Naito, H. Hijikata, T. A. Tsunoyama, N. Hiramoto-Yamaki, R. S. Kasai, Y. M. Shirai, M. S. Miyahara, T. K. Fujiwara, and A. Kusumi. Dynamic regulation of AMPA receptor and Stargazin concentration in the spine in the time scale of 0.1 s to several 100 s; unraveling by single-molecule tracking. The 63rd Annual Meeting of Biophysical Society (BPS19). Baltimore, Maryland, U.S.A. March 5, 2019.

 

ORAL PRESENTATIONS (General)

T. A. Tsunoyama, K. G. N. Suzuki, T. K. Fujiwara, and A. Kusumi. β1 and 3 integrin function in focal adhesion formation and disintegration; unraveling by super-long single-fluorescent molecule tracking. Okayama, Japan. The 56th Annual Meeting of the Biophysical Society of Japan. Okayama, Japan. September 16, 2018.

 

POSTER PRESENTATIONS

P. Zhou, R. S. Kasai, K. M. Hirosawa, T. K. Fujiwara, T. A. Tsunoyama, A. Yudin, and A. Kusumi. Transient hetero-dimerization of opioid receptors (GPCRs) revealed by single-molecule tracking. Cold Spring Harbor Laboratory (CSHL) meeting on “Single Biomolecules”. Cold Spring Harbor, New York, U.S.A. August 30, 2018.

S. Acharya, T .A. Tsunoyama, A. Aladag, I. Meshcheryakova, A. Nakamura, T. K. Fujiwara, and A. Kusumi. Transient dimerization of a postsynaptic cell adhesion molecule neuroligin and its implications in the regulation of trans-synaptic adhesion Molecular Mechanisms of Neuronal Connectivity. Cold Spring Harbor Laboratory (CSHL) meeting on “Molecular Mechanisms of Neuronal Connectivity”. New York, U.S.A. September 25, 2018.

P. Zhou, R. S. Kasai, K. M. Hirosawa, T. K. Fujiwara, T. A. Tsunoyama, A. Yudin, and A. Kusumi. Transient hetero-dimerization of opioid receptors (GPCRs) revealed by single-molecule tracking. The 2019 joint meeting of the American Society for Cell Biology (ASCB) | EMBO Meeting. San Diego, California, U.S.A. December 9, 2018.

T. A. Tsunoyama, R. S. Kasai, K. G. N. Suzuki, T. K. Fujiwara, and A. Kusumi. Super-long single fluorescent-molecule tracking reveals dynamic and tension-dependent anchorage of integrin as the basic mechanism for cell adhesion. The 2019 joint meeting of the American Society for Cell Biology (ASCB) | EMBO Meeting. San Diego, California, U.S.A. December 9, 2018

J. Morise, K. G. N. Suzuki, A. Kitagawa, Y. Wakazono, K. Takamiya, T. A. Tsunoyama, H. Takematsu, A. Kusumi, and S. Oka. Monomers of AMPA-type glutamate receptor subunits diffuse in and out of spines; unraveling by single-molecule tracking. The 63rd Annual Meeting of Biophysical Society (BPS19). Baltimore, Maryland, U.S.A. March 6, 2019.

Y. L. Nemoto, K. Naito, H. Hijikata, T. A. Tsunoyama, N. Hiramoto-Yamaki, R. S. Kasai, and A. Kusumi. Fast and slow turnover of AMPA receptor and stargazin in the spine in the time scale of 0.1 s to several 100 s; unraveling by single molecule imaging. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

P. Zhou, R. S. Kasai, K. M. Hirosawa, T. K. Fujiwara, T. A. Tsunoyama, A. Yudin, and A. Kusumi. Transient hetero-dimerization of opioid receptors (GPCRs) revealed by single-molecule tracking. The 16th International Membrane Research Forum. Okinawa, Japan. March 9, 2019.

A. Yudin, T. K. Fujiwara, T. A. Tsunoyama, and A. Kusumi. Evaluating the permeability across the actin-based compartment barrier. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

S. Acharya, T. A Tsunoyama, A. Aladag, I. Meshceryakova, A. Nakamura, T. K. Fujiwara, and A. Kusumi. Transient dimerization of synaptic cell adhesion molecules neuroligin and neurexin and its implications in the regulation of trans-synaptic adhesion. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

J. Morise, K. G. N. Suzuki, A. Kitagawa, Y. Wakazono, K. Takamiya, T. A. Tsunoyama, H. Takematsu, A. Kusumi, and S. Oka. AMPA receptors in the synapse turnover by monomer diffusion; unraveling by single-molecule tracking. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

H.-C. Lim, and J. Tzuu-Shuh. EBP50 is localized at the blebs and phosphorylated at Serine 347-348 residues in mitotic cells. The 16th International Membrane Research Forum. Okinawa, Japan. March 18, 2019.

 

5. Intellectual Property Rights and Other Specific Achievements

  

Patent Number: 6398055

Inventor: Michio Murata, Nobuaki Matsumori, Hisataka Kinoshita, Akihiro Kusumi, and Ken-ichi Suzuki
Name of Invention: New fluorescently-labeled sphingomyelins and their applications
Patent rights: Osaka University, Akihiro Kusumi, and Ken-ichi Suzuki
Registered date: 14 September 2018
Application date: 15 March 2016

 

6. Meetings and Events

OIST Joint Mini-Symposium

The 16th International Membrane Research Forum

Featuring 2D-3D Meso-Scale Functional Molecular Complexes and Domains in Cellular Membranes
Date: 18 - 20 March 2019  
Venue: Central Building C209
 

Organizers:  

  • Executive Committee for the Membrane Research Forum
  • Standing Committee Members:  Jiro Usukura (Nagoya), Masahiro Sokabe (Nagoya), and AkihiroKusumi (Chair, OIST)
  • OIST Organizers: Keiko Kono and Akihiro Kusumi

Organizing Institutions:

  •   Okinawa Institute of Science and Technology Graduate University (OIST)
  •   Executive Committee for the International Membrane Research Forum

Co-Sponsors (Alphabetical Order):

  • Japan Society for Promotion of Science (JSPS), Grants-in-Aid for Scientific Research (awarded to Keiko Kono and Akihiro Kusumi)
  • The Naito Foundation (awarded to Keiko Kono)
  • Japan Science and Technology Agency (JST), CREST Program of “Creation of  Fundamental for Understanding and Control of Biosystem Dynamics" (Program Director; Prof. Tadashi Yamamoto of OIST)
  • Okabe Team “Understanding synapse dynamics through nanoscale structural analyses”
  • Kono Unit (Membranology Unit) and Kusumi unit (Membrane Cooperativity Unit), OIST
  • OIST Imaging Section, Research Support Division, OIST

Corporate Co-Sponsors (Alphabetical Order):

  •   AIRIX Corp.
  •   Andor Technology Ltd.
  •   Bitplane/Andor Technology Ltd.
  •   Chroma Technology Japan G. K.
  •   Chuo Iryoki CO., LTD
  •   GORYO Chemical, Inc.
  •   HAMAMATSU PHOTONICS K.K.
  •   Inohara Co. & Ltd.
  •   Japan Laser Corporation
  •   JEOL LTD.
  •   Leica Microsystems K.K.
  •   Nikon Instech Co., Ltd.
  •   Nippon Gatan, Gatan Division of Nippon Roper KK
  •   OLYMPUS CORPORATION
  •   OptoSigma
  •   Photron Limited.
  •   PNEUM Co., Ltd
  •   Tokai Hit Co., Ltd.
  •   TOMY OKINAWA NOVO SCIENCE.CO.LTD
  •   WAKENYAKU, Co., Ltd.
  •   Yokogawa Electric Corporation

Meeting Coordinators:

  •   Miwako Tokuda, Hitomi Ohtaki, Amine Betul Nuriseria Aladag, Irina Meshcheryakova,
  •   Aya Nakamura, Hisae Tsuboi, and Hiroko Hijikata
  •   OIST Workshop Section (Hitomi Miyazato, Shoko Nakamatsu, and Chieri Matsuda)
  •   OIST Imaging Section (Bruno Humbel, Shinya Komoto, and Ryo Kanno)

Design and Graphics:

  •   Koji Kanemasa (President and Designer, incomings)

Invited Speakers (in the order of presentations; * indicates a keynote speaker):

  •   Prof. Katharina Gaus, University of New South Wales, Australia*
  •   Prof. Takahiro K. Fujiwara, Kyoto University
  •   Prof. Jiro Usukura, Nagoya University
  •   Prof. Ilya Levental, University of Texas Health Science Center – Houston, U.S.A.*
  •   Prof. Yuji Hara, Kyoto University; AMED-PRIME
  •   Prof. Takeharu Nagai, Osaka University
  •   Prof. Tadashi Yamamoto, OIST
  •   Prof. Barbara Baird, Cornell University, U.S.A.*
  •   Mr.  Koichiro M. Hirosawa, Gifu University
  •   Prof. Anthony Watts, University of Oxford, U.K.*
  •   Dr.  Peng Zhou, OIST
  •   Dr.  Taka A. Tsunoyama, OIST
  •   Prof. Ichiro Maruyama, OIST
  •   Prof. Yves Barral, ETH Zürich, Switzerland*
  •   Prof. Yusuke Hirabayashi, The University of Tokyo
  •   Prof. Harald Stenmark, Institute for Cancer Research, Oslo University Hospital, Norway*
  •   Prof. Michiko Shirane, Nagoya City University
  •   Dr.  Ikuko Koyama-Honda, The University of Tokyo, JST-ERATO
  •   Prof. Issei Mabuchi, The University of Tokyo
  •   Prof. Shindo Asako, Nagoya University
  •   Prof. Masahiro Sokabe, Nagoya University Graduate School of Medicine
  •   Prof. Masato Umeda, Kyoto University
  •   Prof. Roger Nicoll, University of California San Francisco, U.S.A.*
  •   Prof. Hideji Murakoshi, National Institute for Physiological Sciences
  •   Prof. Yasunori Hayashi, Kyoto University Graduate School of Medicine
  •   Prof. Haruhiko Bito, The University of Tokyo Graduate School of Medicine
  •   Dr.  Hiroko Bannai, JST PRESTO / RIKEN-CBS
  •   Ms.  Yuri L. Nemoto, OIST
  •   Dr.  Françoise Coussen IINS, CNRS, France
  •   Prof. Pakorn (Tony) Kanchanawong, National University of Singapore, Singapore*
  •   Prof. Sawako Yamashiro, Kyoto University
  •   Prof. Keiko Kono, OIST
  •   Prof. Satoshi Yoshida, School of International Liberal Studies, Waseda University
  •   Prof. Tsuyoshi Hirashima, Graduate School of Medicine, Kyoto University
  •   Prof. Kandice Levental, University of Texas Health Science Center – Houston, U.S.A.
  •   Dr.  Dragomir Milovanovic, Yale School of Medicine, U.S.A.*
  •   Prof. Tomoyuki Takahashi, OIST
  •   Prof. Bernd Kuhn, OIST
  •   Prof. Makoto Kinoshita, Nagoya University Graduate School of Science
  •   Prof. Daniel Choquet, CNRS-Bordeaux University, France*