2004 Fiscal Year Final Research Report Summary
Analysis of differentially expressed genes in denervated skeletal muscle using micro array technology
| Project/Area Number |
14570619
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | Showa University |
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Principal Investigator |
JIMI Takahiro Showa University, School of medicine, Assistant Professor, 医学部, 助教授 (30196654)
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| Co-Investigator(Kenkyū-buntansha) |
WAKAYAMA Yoshihiro Showa University, School of Medicine, Professor, 医学部, 教授 (40138467)
SHIBUYA Seiji Showa University, School of Medicine, Assistant Preoessor, 医学部, 助教授 (80167444)
HARA Hajima Showa University, School of Medicine, Lecturer, 医学部, 講師 (30241029)
INOUE Msahiko Showa University, School of Medicine, Assistnat, 医学部, 助手 (50286770)
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| Project Period (FY) |
2002 – 2004
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| Keywords | denervation / skeletal muscle / microbeads / DNA array / gene sxpression / molecular chaperone |
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| Research Abstract |
To examine effects of neural factors on gene expression in skeletal muscle, we analyzed differential mRNA expression under denervated condition. Experimentally denervated rat EDL muscle was prepared by removal of the unilateral sciatic nerve, and 72 hours later, the EDL muscle was excised The total RNA was extracted from both normally innervated and the experimentally denervated EDL muscles. As probes, the W firm innervated muscle and that from the denervated Hustle was labeled with Cy5 and fluorescein, respectively. Both probes were combined and competitively hybridized to 0.4 million ready-made DNA microbeads (TakaraBio). On the surface of each DNA microbead, cDNA fragments consisting of one kind of rat skeletal muscle of were fixed. After hybridization, the microbeads were analyzed by a cell sorter. A two-color dot plot indicated the area where differential up-regulation or down-regulation of expressed genes in the denervated condition occurred. The microbeads in the target area were isolated by the cell sorter. More than twenty-five genes isolated were up-regulated in the denervated condition Among them several noticeable genes including heat shock protein 27, alpha B crystallin, and thioredoxin were included. Most of then were classified into the category of proteins for the quality control. The results suggest that various stresses or protein misfolding might occur in the denervated skeletal rustle. More than thirty-seven genes isolated were down-regulated in the denervated condition They contained many metabolic enzymes such as phosphoglycerate mutase and enolase 3. The results suggest that denervation caused disturbance of normal metabolic process of skeletal muscle. Isolated genes were used to make custom-made IINA chips to analyze time-sequential change of expression of these genes. These genes may be useful in understand= neural effects on skeletal rustle.
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