| Project/Area Number |
11460125
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | TOUHOKU UNIVERSITY |
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Principal Investigator |
AKIBA Yukio Tohoku University, Graduate School of Agricultural Science, Professor, 大学院・農学研究科, 教授 (30005631)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Kan Tohoku University, Graduate School of Agricultural Science, Research assistant, 大学院・農学研究科, 助手 (20250730)
高橋 和昭 東北大学, 大学院・農学研究科, 助手 (80183440)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 2000: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥7,600,000 (Direct Cost: ¥7,600,000)
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| Keywords | Skeletal muscle / myoblast / Differentiation / Cloning / MyoD / Myogenin / Chicken / Myo D / myogenin |
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| Research Abstract |
The improvement of meat production may be requested to meet the expanding demands for foods all over the world. In the present study, preparation of chicken myoblast cell line and regulation of gene expression related to the proliferation and differentiation of chicken myoblasts was conducted to construct the base line for promoting the efficient meat production system.
1. Preparation of chicken myoblast cell line
a. Chicken myoblasts could be cloned by the colony formation procedure, while the proliferation could not be maintained for several generations.
b. b. SV40 Large T was transfected to chicken primary myoblasts for potentiating the proliferation activity. The expression of SV40 Large T was transiently appeared in the cell clone.
c. Promoter sequences of specific proteins in chicken muscle cells were subjected for cloning and ligated to plasmid (pd2EGFP-1) which expressed GFP, and then transfected to the chicken primary myoblasts. The analysis is currently continued and the preparat
… More
ion of chicken muscle cell line is scheduled with aid of this procedure.
2. Screening of factors regulating proliferation and differentiation in chicken myoblasts
a. In the chicken myoblast culture, MyoD and Myogenin mRNA expression were peaked at 12 and 2h hr, respectively and they were followed by gradual decrease.
b. Myosin, CPK activity and MyD mRNA expression increased by addition of prostaglandin E2 (PGE2), suggesting that PGE2 is a factor stimulating the differentiation of muscle cells.
c. Proliferation in the myocyte growth stage and fusion of myoblasts in the terminal differentiation was stimulated by addition of beta-hydroxybutyrate (BTB), a ketone body, in the myocyte culture and these effects were modified by the amount of energy source present in the medium.
d. Residual yolk present in the abdomen of post-hatched chicks appears to contain a plenty of proteins rich in metabolic function. Addition of the proteins extracted from the residual yolk was no influence on the muscle cell differentiation at the low concentration (1-100 ug) but induced the cell death at the high concentration (1 or 3 mg). Less
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