| 研究実績の概要 |
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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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
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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