2013 Fiscal Year Final Research Report
Exhausive analysis of synchronized factors regulating racing ability in thoroughbred horse
Project/Area Number |
23700722
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Research Category |
Grant-in-Aid for Young Scientists (B)
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Allocation Type | Multi-year Fund |
Research Field |
Sports science
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Research Institution | Yamaguchi University |
Principal Investigator |
YAMANO Seiko 山口大学, 大学研究推進機構, 技術職員 (00448276)
|
Project Period (FY) |
2011 – 2013
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Keywords | サラブレッド / 骨格筋 / 体内環境シンクロナイズ因子 |
Research Abstract |
To identify the differentiation factors released in an ischemia cell model using the myoblast-derived cell line, H9c2 cells to the skeletal muscle cells through MyoD, the cell supernatant was separated by ion exchange column, and was examined by MyoD transcriptional activity. The transcriptional activity was undetected in the supernatant. We have carried out proteomic analysis of proteins released from cells during ischemic hypoxia and reoxygenation using H9c2 cells. the cultured supernatant was concentrated more than 10,000-fold by column chromatographies, and the proteins were separated by SDS-PAGE. Proteomic data using nano-LC-Q-Tof-MSMS showed more than 100 proteins specifically released from cells during hypoxic stress compared with control cells. The identified 4 factors were cloned, and introduced in Halo-tag. The cDNAs were transfected in HeLa cells, and partially purified by affinity column. In near future, the factors would be examined in MyoD transcriptional activity.
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