Optical detection of the molecular processes underlying hippocampal LTP

Summary

Principal Investigator: Yasunori Hayashi
Abstract: Synaptic plasticity has been suggested to be a cellular counterpart for learning and memory. However, it is practically impossible to visually monitor synapses actually undergoing synaptic plasticity at a given memory paradigm in a given neuronal network. This project proposal, written in response to the program announce- ment "Developing Novel Genetic Methods For Mapping Functional Neuronal Circuits and Synaptic Change", describes the development of a technology for visualizing synapses that are undergoing synaptic plasticity in neurons in living animal. This goal will be accomplished by a combination of two technologies: fluorescence resonance energy transfer (FRET) and two-photon laser scanning microscopy. We will first develop a FRET-based construct for optically measuring the CaMKII activity and actin polymerization/depolymerization equilibrium, which will allow non-destructive optical detection of CaMKII activation in intact neurons. Since the enzymatic activity of CaMKII is constitutively enhanced after the induction of NMDA receptor-mediated syn- aptic plasticity and this activity is required for maintaining synaptic potentiation, we expect that the activation of CaMKII will be a good indicator of synaptic plasticity. In contrast, in a work which we published, we found actin polymerization/depolymerization equilibrium can be detected with FRET and it follow LTP and LTD respec- tively. To detect FRET at the synaptic structure, we will take advantage of a two-photon microscope. We will then test the feasibility of our strategy by expressing the construct in neurons and induce synaptic plasticity by either high-speed ionophoretic stimulation of individual synapses or local electrical stimulation or combined with detection of structural plasticity. Finally, we will generate a transgenic animal expressing this construct. The barrel cortex of the resultant animal will be observed with the two-photon microscope. Paradigms known to induce synaptic plasticity in this structure such as sensory deprivation will be tested to see whether synaptic plasticity is reflected by FRET. In summary, our technology will provide a unique system for detecting NMDAR-mediated synaptic plas- ticity with spatial resolution at the single synapse level on a sub-second time scale. This technique, in com- bination with technologies currently under development in other laboratories, such as in vivo two-photon im- aging in freely moving animals and deep-structure two-photon microscope imaging with a relay lens, will pro- vide a versatile system for monitoring synaptic plasticity that cannot be achieved with existing experimental systems. In the future, this technique could be applied to higher mammals, such as macaque monkeys, after viral or transgenic introduction of our reporter construct. Itmay be possible to have monkeys perform a task and observe synaptic plasticity in the brain during learning of the task.
Funding Period: ----------------2006 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. pmc The role of CaMKII as an F-actin-bundling protein crucial for maintenance of dendritic spine structure
    Ken ichi Okamoto
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Proc Natl Acad Sci U S A 104:6418-23. 2007
  2. ncbi Structural and molecular remodeling of dendritic spine substructures during long-term potentiation
    Miquel Bosch
    RIKEN MIT Neuroscience Research Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA Electronic address
    Neuron 82:444-59. 2014
  3. pmc Application of FRET probes in the analysis of neuronal plasticity
    Yoshibumi Ueda
    Brain Science Institute, RIKEN Wako, Saitama, Japan
    Front Neural Circuits 7:163. 2013
  4. pmc PIP₃ regulates spinule formation in dendritic spines during structural long-term potentiation
    Yoshibumi Ueda
    Brain Science Institute, RIKEN, Wako, Saitama 351 0198, Japan
    J Neurosci 33:11040-7. 2013
  5. ncbi The Ca2+ and Rho GTPase signaling pathways underlying activity-dependent actin remodeling at dendritic spines
    Takeo Saneyoshi
    Brain Science Institute, RIKEN, Wako, Saitama, Japan
    Cytoskeleton (Hoboken) 69:545-54. 2012
  6. pmc Activation of sodium-dependent glutamate transporters regulates the morphological aspects of oligodendrocyte maturation via signaling through calcium/calmodulin-dependent kinase IIβ's actin-binding/-stabilizing domain
    Zila Martínez-Lozada
    Department of Anatomy and Neurobiology, Virginia Commonwealth University Medical Center, Richmond, Virginia Departamento de Toxicología, Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional, Mexico D F, Mexico
    Glia 62:1543-58. 2014
  7. ncbi Structural plasticity of dendritic spines
    Miquel Bosch
    The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Curr Opin Neurobiol 22:383-8. 2012
  8. pmc Genetically encoded probe for fluorescence lifetime imaging of CaMKII activity
    Showming Kwok
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue 46 4243A, Cambridge, MA 02139, USA
    Biochem Biophys Res Commun 369:519-25. 2008
  9. ncbi Genetic ablation of NMDA receptor subunit NR3B in mouse reveals motoneuronal and nonmotoneuronal phenotypes
    Stephan Niemann
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA
    Eur J Neurosci 26:1407-20. 2007
  10. ncbi Motoneuron-specific NR3B gene: no association with ALS and evidence for a common null allele
    S Niemann
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA
    Neurology 70:666-76. 2008

Scientific Experts

  • Yasunori Hayashi
  • Miquel Bosch
  • Yoshibumi Ueda
  • Ken ichi Okamoto
  • Showming Kwok
  • Takeo Saneyoshi
  • Zila Martínez-Lozada
  • Hitomi Matsuno
  • Mriganka Sur
  • Dan Ohtan Wang
  • Amanda F Mower
  • Kenichi Okamoto
  • Mariko Kato Hayashi
  • S Niemann
  • Radhakrishnan Narayanan
  • Stephan Niemann
  • Shiping Zou
  • Pamela E Knapp
  • Christopher T Waggener
  • Arturo Ortega
  • Jorge Castro
  • Karam Kim
  • Babette Fuss
  • Akimitsu Okamoto
  • Shuji Ikeda
  • Akiko Nakamura
  • Hiroyuki Yanagisawa
  • Ania K Majewska
  • Hongbo Yu
  • Chiara Verpelli
  • Rui Ming Xu
  • Huilin Li
  • Marissa H Stearns
  • Carlo Sala
  • Chunyan Tang
  • Susana A Sanchez
  • B Hosler
  • M J Churchill
  • R H Brown
  • J E Landers
  • P Sapp
  • Claudia Lee
  • K K Kidd
  • B T Lahn
  • Enrico Gratton
  • W C Speed
  • Theodore L Hazlett
  • Shigeyoshi Itohara
  • Jeremy M Shefner
  • Hiroaki Kanki
  • Kazuyoshi Murata
  • Robert H Brown
  • Roderick T Bronson
  • Yasuyuki Fukui
  • Sang H Lee
  • Keizo Takao
  • Tsuyoshi Miyakawa
  • Masahiro Fukaya
  • Michael J Churchill
  • Meri N Hynynen
  • Masahiko Watanabe

Detail Information

Publications16

  1. pmc The role of CaMKII as an F-actin-bundling protein crucial for maintenance of dendritic spine structure
    Ken ichi Okamoto
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Proc Natl Acad Sci U S A 104:6418-23. 2007
    ..This finding indicates that CaMKII serves as a central signaling molecule in both functional and structural changes during synaptic plasticity...
  2. ncbi Structural and molecular remodeling of dendritic spine substructures during long-term potentiation
    Miquel Bosch
    RIKEN MIT Neuroscience Research Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA Electronic address
    Neuron 82:444-59. 2014
    ..Our findings show how and when spine substructures are remodeled during LTP and explain why synaptic plasticity rules change over time. ..
  3. pmc Application of FRET probes in the analysis of neuronal plasticity
    Yoshibumi Ueda
    Brain Science Institute, RIKEN Wako, Saitama, Japan
    Front Neural Circuits 7:163. 2013
    ....
  4. pmc PIP₃ regulates spinule formation in dendritic spines during structural long-term potentiation
    Yoshibumi Ueda
    Brain Science Institute, RIKEN, Wako, Saitama 351 0198, Japan
    J Neurosci 33:11040-7. 2013
    ..Subspine imaging revealed that when sLTP was induced in a single spine, PIP₃ accumulates in the spinule whereas PIP₃ concentration in the spine decreased...
  5. ncbi The Ca2+ and Rho GTPase signaling pathways underlying activity-dependent actin remodeling at dendritic spines
    Takeo Saneyoshi
    Brain Science Institute, RIKEN, Wako, Saitama, Japan
    Cytoskeleton (Hoboken) 69:545-54. 2012
    ..Here, we will review recent studies examining the signaling pathways underlying activity-dependent actin remodeling at excitatory postsynaptic neurons...
  6. pmc Activation of sodium-dependent glutamate transporters regulates the morphological aspects of oligodendrocyte maturation via signaling through calcium/calmodulin-dependent kinase IIβ's actin-binding/-stabilizing domain
    Zila Martínez-Lozada
    Department of Anatomy and Neurobiology, Virginia Commonwealth University Medical Center, Richmond, Virginia Departamento de Toxicología, Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional, Mexico D F, Mexico
    Glia 62:1543-58. 2014
    ....
  7. ncbi Structural plasticity of dendritic spines
    Miquel Bosch
    The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Curr Opin Neurobiol 22:383-8. 2012
    ..Here we review the pioneering microscopic studies examining the structural plasticity of spines and propose how changes in actin treadmilling might regulate spine morphology...
  8. pmc Genetically encoded probe for fluorescence lifetime imaging of CaMKII activity
    Showming Kwok
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue 46 4243A, Cambridge, MA 02139, USA
    Biochem Biophys Res Commun 369:519-25. 2008
    ..These FLIM versions of Camui could be useful for elucidating the function of CaMKII both in vitro and in vivo...
  9. ncbi Genetic ablation of NMDA receptor subunit NR3B in mouse reveals motoneuronal and nonmotoneuronal phenotypes
    Stephan Niemann
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA
    Eur J Neurosci 26:1407-20. 2007
    ..This work is the first reporting of the functional significance of NR3B in vivo and may give insight into the contribution of genetic variability of NR3B in the phenotypic heterogeneity among human population...
  10. ncbi Motoneuron-specific NR3B gene: no association with ALS and evidence for a common null allele
    S Niemann
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA
    Neurology 70:666-76. 2008
    ..We tested whether genetic dysfunction of GRIN3B is implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS)...
  11. ncbi Visualization of F-actin and G-actin equilibrium using fluorescence resonance energy transfer (FRET) in cultured cells and neurons in slices
    Ken ichi Okamoto
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 46 4243A, 43 Vassar Street, Cambridge, Massachusetts 02139, USA
    Nat Protoc 1:911-9. 2006
    ..Our protocol provides the first direct method for looking at the dynamic equilibrium between F-actin and G-actin in intact cells...
  12. ncbi The roles of CaMKII and F-actin in the structural plasticity of dendritic spines: a potential molecular identity of a synaptic tag?
    Kenichi Okamoto
    Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada
    Physiology (Bethesda) 24:357-66. 2009
    ..The status of F-actin, regulated by CaMKII, determines the postsynaptic protein binding capacity and thus may act as a synaptic tag that consolidates LTP...
  13. pmc The postsynaptic density proteins Homer and Shank form a polymeric network structure
    Mariko Kato Hayashi
    RIKEN MIT Neuroscience Research Center, The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Cell 137:159-71. 2009
    ..We propose that the Homer-Shank complex serves as a structural framework and as an assembly platform for other PSD proteins...
  14. pmc Experience-dependent regulation of CaMKII activity within single visual cortex synapses in vivo
    Amanda F Mower
    The Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Proc Natl Acad Sci U S A 108:21241-6. 2011
    ..Synapses that are eliminated in deprived eye domains have low basal CaMKII activity, implying a protective role for activated CaMKII against synapse elimination...
  15. ncbi Roles of neuronal activity-induced gene products in Hebbian and homeostatic synaptic plasticity, tagging, and capture
    Yasunori Hayashi
    Brain Science Institute, RIKEN, 2 1 Hirosawa, Wako, Saitama 351 0198, Japan
    Adv Exp Med Biol 970:335-54. 2012
    ....
  16. pmc A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes
    Dan Ohtan Wang
    Institute for Integrated Cell Material Sciences, Kyoto University, Kyoto 606 8501, Japan
    RNA 18:166-75. 2012
    ..We also demonstrate detection of telomeric and centromeric DNA on metaphase mouse chromosomes. The simplicity of the ECHO-FISH method will likely accelerate cytogenetic and gene-expression analysis with high resolution...