2016 Fiscal Year Research-status Report
リン酸化プロテオミクスで拓く新規Sleepy遺伝子の睡眠恒常性維持機構への関与
| Project/Area Number |
16K16639
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| Research Institution | University of Tsukuba |
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Principal Investigator |
Wang Zhiqiang 筑波大学, 国際統合睡眠医科学研究機構, 研究員 (00762189)
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| Project Period (FY) |
2016-04-01 – 2018-03-31
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| Keywords | phosphoproteome / sleep deprivation / Sleepy / sleep need / synaptic proteins / NOS1 |
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| Outline of Annual Research Achievements |
In last year’s study, we performed quantitative phosphoproteomic studies of whole mouse brains from two models of sleep/wake perturbation. A combined proteome and phosphoproteome data for 9,410 mouse proteins and 4, 5000 phosphopeptides were examined. Analysis of ad-lib slept, sleep-deprived, and recovery slept mice revealed that sleep and waking drove hypo- and hyper-phosphorylation of brain proteome. In Sleepy mice, a gain-of-function mutation of SIK3 caused constitutively higher sleep need and phosphoproteome imbalance mimicking sleep-deprived condition. The intracerebroventricular (i.c.v.) injection-based and AAV overexpression-based EEG/EMG sleep recording system have been established, the Nos1 inhibitor treatment and overexpress Nos1 on Sleepy are ongoing.
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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 have finished phosphoproteomic studies of whole mouse brains from two models of sleep/wake perturbation. The Bioinformatics analysis of the proteome and phosphoproteome data almost completed and generated several new insights. And we found more interesting Sleepy substrates. Two sleep study system, i.c.v. and AAV overexpression, have been established to test the functions of those substrates.
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| Strategy for Future Research Activity |
In new year, we will investigate the function of Sleepy substrates, such as NOS1 through pharmacological and AAV overexpression approaches. Moreover, a manuscript for bioinformatics analysis of the proteome and phosphoproteome data will be prepared.
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| Causes of Carryover |
A new experiment to test new AAV virus system will be done in FY2017.
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| Expenditure Plan for Carryover Budget |
This money will be used to buy new type AAV virus plasmids and packaging reagents like PEI.
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