2014 Fiscal Year Annual Research Report
| Project/Area Number |
14F04760
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
合田 裕紀子 独立行政法人理化学研究所, 脳科学総合研究センター, チームリーダー (40614897)
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| Co-Investigator(Kenkyū-buntansha) |
CHIPMAN Peter 独立行政法人理化学研究所, 脳科学総合研究センター, 外国人特別研究員
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| Project Period (FY) |
2014-04-25 – 2016-03-31
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| Keywords | 国際研究者交流 |
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| Outline of Annual Research Achievements |
Two variants of the pseudo-typed, glycoprotein-deleted monosynaptic rabies virus were prepared: one expressing mCherry and another expressing GFP. Using hippocampal slice cultures, single cell electroporation was then performed to successfully direct these viruses to infect single CA1 neurons and to promote the restricted retrograde spread of the virus to the presynaptic partners. The health of virus-infected cells are currently being assessed by dual patch-clamp recordings. Also underway are pilot triple patch-clamp recording experiments and testing the induction of plasticity in one of the two convergent inputs.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Apart from the occasional setbacks caused by unhealthy cultures, experiments are progressing as planned.
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| Strategy for Future Research Activity |
Spontaneously active cell assemblies will be identified in hippocampal slice cultures using genetically encoded calcium indicators and electrophysiology. Specifically, the rabies virus will be engineered to express GCaMP6. The activity of presynaptic neurons will be recorded to determine the presence of “cell assemblies” as defined by presynaptic neurons that show spontaneous synchronous activity that is coincident with the activity of the postsynaptic neuron. Patch clamp recording experiments from a postsynaptic neuron and its two presynaptic neurons will be performed to determine if presynaptic inputs belonging to separate cell assemblies show distinct synaptic properties. In order to increase the yield of successful recordings, three presynaptic neurons will be patch-clamped simultaneously along with a postsynaptic neuron. For this purpose, another micromanipulator is requested for Yr2 of the project.
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