| Project/Area Number |
10470232
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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 |
Metabolomics
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| Research Institution | The University of Tokushima |
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Principal Investigator |
KUWAJIMA Masamichi The Tokushima University, School of Medicine, Associate Professor, 医学部, 助教授 (00205262)
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| Co-Investigator(Kenkyū-buntansha) |
MIZUNO Akira The Tokushima University, Medicine School Hospital, Research Associate, 医学部・附属病院, 助手 (80219641)
NOMA Yoshihiko The Tokushima University, Medicine School Hospital, Assistant Professor, 医学部・附属病院, 講師 (10218349)
ITO Michinori The Tokushima University, School of Medicine, Associate Professor, 医学部, 助教授 (40211057)
TANAKA Toshihiro University of Tokyo, Human Genome, Center, Instructor, 医科学研究所・ヒトゲノム解析センター, 助手 (50292850)
MURAKAMI Takashi The Tokushima University, School of Medicine, Instructor, 医学部, 助手 (40210009)
篠原 康雄 徳島大学, 薬学部, 助教授 (60226157)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 1999: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1998: ¥9,900,000 (Direct Cost: ¥9,900,000)
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| Keywords | carnitine / carnitine transporter / hypoglycemia / JVS mouse / octn2 / lactate / pyruvate / NaィイD1+ィエD1 dependency / 心筋細胞 / OCTN^2 / ポジショナルクローニング / β-酸化 / JVS マウス / カルニチントランスポーター |
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| Research Abstract |
Juvenile Visceral Steatosis (JVS) mouse, which we reported in 1991, serves as an animal model of primary carnitine deficiency. Because this mouse is suffered from severe hypoglycemia, we analyzed the cause and mechanism The summary of the results is as follows ;
1. Assay of carnitine transport activity
We have conducted a kinetic analysis using fibroblasts derived from normal, heterozygous, and homozygous JVS mice and found that the high-affinity carnitine transporter, which shows NaィイD1+ィエD1 dependency, is defective in homozygous JVS mice. Moreover, a gene dose-dependent decrease of carnitine transport activity was found in heterozygous JVS mice.
2. Analysis of candidate gene
As a human OCTN2 gene encoding a sodium-dependent carnitine cotranspoter was isolated, we isolated the mouse octn2 gene and screened for its mutation in the JVS mouse. DNA sequencing analysis disclosed a missense mutation from CTG (Leu) to CGG (Arg) at codon 352 located within the sixth transmembrane domain of octn2. This amino acid replacement possibly causes the conformational change of the protein that leads to dysfunction of the gene product.
3. Analysis of hypoglycemia
The glucose level of JVS mouse at ad lib feeding was lower than that of normal control. After the prolonged starvation, the glucose level of JVS mouse was significantly decreased at 60 hours in comparison with normal control. The insulin level was not changed between JVS and normal control mouse. The levels of pyruvate and lactate were decreased. This decreased was thought as a cause of hypoglycemia.
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