2016 Fiscal Year Annual Research Report
アストロサイトとシナプスのコンタクトダイナミックスの機構
| Project/Area Number |
15H04280
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
合田 裕紀子 国立研究開発法人理化学研究所, 脳科学総合研究センター, チームリーダー (40614897)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Keywords | synaptic transmission / astrocyte |
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| Outline of Annual Research Achievements |
Towards the overall aim of characterizing the dynamic interaction between astrocytes and synapses, to date, we have constructed astrocyte-specific probes to visualize and manipulate astrocyte activity while monitoring synaptic transmission. We first confirmed the expression of probes, including mCherry, ArchT, ChR2 and LifeAct-GCaMP, in astrocytes in dissociated hippocampal cultures. Subsequently, we prepared AAV vectors of individual probes; AAV was injected into mice brains, and the expression was evaluated in acute and fixed hippocampal slices. Specificity of astrocyte expression varied amongst different versions of the hGFAP promoter across hippocampal subregions such that care was needed in assessing synaptic transmission within the domain of the targeted astrocytes. In separate experiments, pharamcologically blocking gap junctions, which had been previously implicated in affecting synapse-astrocyte structural contacts, modified synaptic transmission in a manner similar to when astrocyte signaling was blocked. In order to determine the molecular basis of the influence of gap junction inhibitor on synapse function, we prepared astrocyte expression vectors for connexins 30 and 43 that had been reported to be present in astrocytes. These probes will be tested in this coming year.
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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
We experienced some delays in constructing various vectors for astrocyte-specific expression, which was caused by the unexpected delay in H27 in hiring of the personnel. We made substantial effort in catching up, and the project is running smoothly at present and no further delays are expected.
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| Strategy for Future Research Activity |
We will further characterize astrocyte filopodia dynamics in relationship to individual synapses while also evaluating the effects of astrocyte-expressed connexins that form gap junctions in affecting astrocyte-synapse dynamics.
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Research Products
(3 results)
