Akemi Kishi
Takaaki Fujii

Our method at JoVE

Connecting synaptic plasticity to memory using Drosophila feeding circuit and Drosophila embryonic neuromuscular junction.

My lab is trying to ask a fundamental question gHow do we remember?h by studying synaptic physiology in the fruit fly, Drosophila. I believe that memory is stored as a series of neurons connected through strengthened synapses, in which sequential firing of the neurons allows recall of specific events (Fig. 1). Therefore, understanding mechanisms of synaptic modification is a key to understanding mechanisms underlying memory. Taking advantage of a combination of synaptic physiological methods on highly plastic Drosophila embryonic neuromuscular synapses (neuromuscular junction, NMJ) and sophisticated Drosophila genetics, we have proposed a novel hypothesis, glocal feedback modelh as a potential molecular and cellular basis of memory formation (Yoshihara et al., 2005, Science 310: 858-863; Fig. 2). In this model, I postulate that mutual intensification between presynaptic and postsynaptic cells by a positive feedback loop at single synapses keeps individual synapses potentiated, leading to eventual morphological change and perpetually strengthened synapses, storing memory. In my lab, we are testing this working hypothesis to answer the question ghow do we remember?h, by a novel approach using a pair of feeding command neurons, Feeding neuron (Fdg neuron; Fig. 3).  We have performed behavioral screening on NP lines (Yoshihara and Ito, 2000, Drosoph. Inf. Serv., 83, 199-202) to establish a new system to connect synaptic plasticity to memory, and finally identified the Fdg neuron (Flood, Iguchi et al., 2013, Nature, 499: 83-87), which is ideal for neurophysiological analysis of classical conditioning demonstrated by Ivan Pavlov (Fig. 4) to connect synaptic plasticity to memory mechanism. We are now establishing novel protocols of Pavlovian conditioning to gwitnessh memory formation as real-time synaptic change in an experimental system I have devised for simultaneous observation of the brain and behavior of a single fruit fly (Yoshihara, 2012, JoVE; 62, 3625; Fig. 5). Through these novel approaches using Drosophila allowing us to perform comprehensive genetic analyses, we are trying to understand basic principle on molecular and cellular mechanism of memory formation.

We are continuing detailed analysis on synaptic plasticity of neuromuscular synapses as a simple cellular model for comparison with central synapses in the brain (this study was supported by NIH R01 grant of U.S.A.). In addition to the plasticity projects, we are also studying basic mechanism of synaptic transmission, which is not fully understood yet.  For these synaptic transmission projects, we are taking advantage of Drosophila embryonic neuromuscular synapses, which is suitable for quantitative synaptic physiology even in lethal mutants, as shown in our previous works on function of synaptic molecules (Yoshihara et al., 1999,  J. Neurosci. 19: 2432-2441; Yoshihara et al., 2000,  J. Neurosci. 20: 8315-8322; Yoshihara and Littleton, 2002 Neuron, 36, 897-908; Yoshihara et al., 2010, Proc. Natl. Acad. Sci. USA, 107, 14869-14874).  We are collaborating with the lab of Prof. J. Troy Littleton at MIT for these neuromuscular projects.

Principal Investigator
Moto lab

Acknowledgements

We greatly thank for generous support to Moto lab by Dr. Iwao Hosako, the Director General of the Advanced ICT Research Institute, Dr. Kazuhiro Oiwa, the Distinguished Researcher of the National Institute of Information and Communications Technology, and Dr, Hiroaki Kojima, the Director of Bio ICT Laboratory.

Motojiro Yoshihara, Ph.D.

Drosophila Synaptic plasticity/memory team

Our strategy in Japanese

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Lab News
Yumi Nakajima (shared)
Postdoctoral fellows
Akira Sakurai (Researcher)@

(Selected publications)

 

Original papers

Flood, T., Gorczyca, M., White, B., Ito, K. and Yoshihara, M.* A large-scale behavioral screen to identify neurons controlling motor programs in the Drosophila brain. 

G3: Genes, Genomes, Genetics   (2013) 3: 1679-1637. *Corresponding author.

Flood, T.˜, Iguchi, S. ˜, Gorczyca, M. ˜, White, B., Ito, K. and Yoshihara, M. * 

A single pair of interneurons commands the Drosophila feeding motor program.

Nature (2013) 499: 83-87. *Corresponding author. ˜Equal contribution

Korkut, C, Li, Y, Koles, K, Brewer, C, Ashley, J, Yoshihara, M, Budnik, V.

Regulation of postsynaptic retrograde signaling by presynaptic exosome release.

Neuron  (2013) 77: 1039-46.

Yoshihara, M.

Simultaneous recording calcium signals from identified neurons and feeding behavior in Drosophila melanogaster. Journal of Visualized Experiments  (2012) 62: e3625, DOI: 10.3791/3625.

Yoshihara, M., * Guan, Z. and Littleton , J.T.

Differential regulation of synchronous versus asynchronous neurotransmitter release by the C2 domains of synaptotagmin 1.

Proc. Natl. Acad. Sci. USA  (2010) 107: 14869-14874  *Corresponding author.

Yoshihara, M., * Adolfsen, B., Galle, K.T. and Littleton, J.T.

Retrograde signaling by Synaptotagmin 4 induces presynaptic release and synapse-specific growth.

Science  (2005) 310: 858-863  *Corresponding author.

 

Adolfsen, B., Saraswati, S., Yoshihara, M. and Littleton, J.T.

Synaptotagmins are trafficked to distinct subcellular domains including the postsynaptic compartment.

J. Cell Biol. (2004) 166: 249-260

Lee, W.C., Yoshihara, M. and Littleton, J.T.

Cytoplasmic aggregates trap polyglutamine-containing proteins and block axonal transport in a Drosophila model of Huntington's disease. 

Proc. Natl. Acad. Sci. USA (2004) 101: 3224-3229

Reickhof, G.E., Yoshihara, M., Guan, Z. and Littleton, J.T.

Presynaptic N-type calcium channels regulate synaptic growth. 

J. Biol. Chem. (2003) 278: 41099-41108

Yoshihara, M. * and Littleton, J.T.

Synaptotagmin I functions as a calcium sensor to synchronize neurotransmitter release. 

Neuron  (2002) 36: 897-908  *Corresponding author.

Hayashi, S., Ito, K. Sado, Y., Taniguchi, M., Akimoto, A., Takeuchi, H., Aigaki, T., Matsuzaki, F., Nakagoshi, H., Tanimura, T., Ueda, R., Uemura, T., Yoshihara, M. and Goto. S. (2002) GETDB, a database compiling expression patterns and molecular locations of a collection of Gal4 enhancer traps. Genesis 34: 58-61.

 Takasu-Ishikawa, E.*, Yoshihara, M.*, Ueda, A., Rheuben, M. B., Hotta, Y. and Kidokoro, Y.

Screening for synaptic defects revealed a locus involved in presynaptic and postsynaptic functions in Drosophila  embryos.

J. Neurobiol. (2001) 48: 101-119  * Equal contribution.

Yoshihara, M., Suzuki, K. and Kidokoro, Y.

Two independent pathways mediated by cAMP and protein kinase A enhance spontaneous transmitter release at Drosophila neuromuscular junctions.

J. Neurosci. (2000) 20: 8315-8322

Yoshihara, M., and Ito, K.

Improved Gal4 screening kit for large-scale generation of enhancer-trap strains. 

Dros. Inf. Ser. (2000) 83: 199-202.

Yoshihara, M., Ueda, A., Zhang, D., Deicher, D. L., Schwarz, T.L. and Kidokoro, Y.

Selective effects of neuronal-synaptobrevin mutations on transmitter release evoked by sustained versus transient Ca2+ increases and by cAMP.

J. Neurosci. (1999) 19: 2432-2441

Yoshihara, M., Rheuben, M. B. and Kidokoro, Y.

Transition from growth cone to functional motor nerve terminal in Drosophila embryos.

J. Neurosci. (1997) 17: 8408-8426

Review in English

Yoshihara M. *, Ito K.

Acute genetic manipulation of neuronal activity for the functional dissection of neural circuits-a dream come true for the pioneers of behavioral genetics.

J. Neurogenet. 26 (2012) 43-52. *Corresponding author.

Yoshihara, M. * and Montana, E.S.

The Synaptotagmins:  Calcium Sensors for Vesicular Trafficking.

The Neuroscientist 10 (2004) 566-574. *Corresponding author.

Yoshihara, M., Adolfsen, B. and Littleton, J.T.

Is synaptotagmin the calcium sensor? 

Current Opinions in Neurobiology 13 (2003) 315-323.

Yoshihara, M., Ensminger , A. and Littleton, J. T.

Neurobiology and the Drosophila genome.

Functional & Integrative Genomics 1 (2001) 235-240.

Review in Japanese

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We are trying to undersand basic principle of memory formation by connecting synaptic plasticity to memory at this Feeding (Fdg) neuron.

Technical stuffs
Former members at UMass or MIT
Akira Sakurai, Postdoctoral fellow

Moto lab moved from@U.S, and@restarted at Motofs home town, Kobe, Japan, in the spring of 2014 in the bio ICT laboratory of the National Institute of Information and Communications technology ("NICT Kobe").

In June, 2014, Nakajima-san started fly food cooking!

Moto taught as an instructor (a 3-hour lecture in the morning and a lab course in the whole afternoon) in the Cold Spring Harbor Course, Neurobiology of Drosophila in July (http://meetings.cshl.edu/courses/2014/c-dros14.shtml).

Moto gave a plenary lecture at International Congress of Neuroethology on July 31st, 2014.

Akira, who had been working in the US Moto lab, rejoined the lab at Kobe on August 1st, 2014 as a researcher to continue the memory project.

Moto gave a plenary lecture at Synapse Workshop at National Institute for Physiological Sciences on December 2nd, 2014.

Akira made a poster presentation at Synapse Workshop at National Institute for Physiological Sciences on December 2nd, 2014.

Moto gave a talk in a Symposium at the Center for Novel Science Initiative in the National Institute of Natural Science on January 17th, 2015.

Moto gave a talk for 1048th Life Science Seminar at University of Tokyo on July 10th, 2015.

Akira made a poster presentation at the 38th Annual Meeting of the Japan Neuroscience Society on July 29th, 2015.

Akira made a poster presentation at Cold Spring Harbor Lavoratory meeting "Neurobiology of Drosophila" on October 2nd, 2015.

Moto gave a talk in the Advanced ICT symposium at Tokyo Big Sight on January 27th, 2016


Moto gave a talk at Engineering Science in Osaka University on October 24th, 2016.

Moto gave a presentation and served as a chairperson at Society for Neuroscience in San Diego on November 11th, 2016.

Moto Yoshihara, principal investigator.

Michael Gorczyca, Postdoctoral fellow
Shinya Iguchi, graduate student (present, UC Berkeley)
Tom Flood, graduate student
Louis Watanabe, summer student (present, University of Alabama)
Sayaka Yokoyama, visiting student from Tohoku University
Alicia Taylor, Coop student from Northeastern University
Rizwana Seeham, summer student from Boston University
Tao Xu, lab technician
at University of Massachusetts Medical school

Moto Yoshihara received his B. Sc. (1987) from the Department of Zoology at the University of Tokyo and Ph.D. (1992) from the Department of Biological Chemistry at the University of Tokyo, Japan. He received a Human Frontiers Science Program fellowship to do postdoctoral work at the City of Hope, California in the laboratory of Kazuo Ikeda. After working as a visiting scientist at the Massachusetts Institute of Technology, he joined the Department of Neurobiology at the University of Massachusetts Medical School as a faculty member in 2006.  In 2013, he left University of Massachusetts Medical School to work as a visiting professor at MIT, being supported by NIH grant, R01.  In 2014, he moved back to Japan, and joined National Institute of Information and Communications Technology as a Senior Research Scientist (PI).

Shinya and Moto, the main force for publication of the Feeding neuron paper in the announce from UMass when our paper appeared on Nature
(http://www.umassmed.edu/news/news-archives/2013/06/neurons-control-feeding-behavior/). 

Contact; Moto Yoshihara, Ph.D.
motojiro@nict.go.jp
Memory Neurobiology Project
Advaced ICT Research Institute ("NICT Kobe")
National Institute of Information and Communiocations Technology,
588-2 Iwaoka, Nishi-ku
Kobe, Japan