Quantitative abnormalities and molecular biological investigation of the ADP/ATP carrier protein in J-2-N cardiomyopathic hamsters

J-2-N 心肌病仓鼠 ADP/ATP 载体蛋白的定量异常和分子生物学研究

• 批准号: 04670559
• 负责人: KATO Mitsutoshi
• 金额: $ 1.34万
• 依托单位: Jikei University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670559/
• 关键词: ADP; ATP carrier protein; cardiomyopathic hamster; mitochondria; ATP carrier protein; Cardiomyopathy; J‐2‐N cardiomyopathic hamster

The ADP/ATP carrier protein(AAC), located in the mitochondrial inner membrane, plays an important role in the mitochondrial energy metabolism. ATP is transported from matrix to cytoplasm by AAC and at the same time ADP is transported inversely. Cloning was performed using myocardium from Bio14.6 hamsters, J-2-N cardiomyopathic hamsters bred in our laboratory, and Golden hamster. The AAC contest was significantly decreased in J-2-N hamsters as compared with Golden hamsters. cDNA of AAC was cloned from cDNA library of the human fibroblast by PCR.The human cDNA was then used to screen Golden hamster AAC cDNA, which was used as a probe in the latter steps. Genomic DNA was prepared from cardiac muscle of the each strain hamster and DNA libraries were constructed. From these libraries, we have succeeded in obtaining genomic DNA of cardiac AAC in hamster. Also RNA dot blot hybridization analysis was performed. The highest mRNA level for AAC was observed in Golden hamster followed by J-2-N with high serum CK, J-2-N with lower serum CK and Bio14.6. These results suggest that decreased mRNA levels of AAC is one of the mechanisms which explain the abnormalities of cardiac metabolism in J-2-N cardiomyopathic hamsters.

ADP/ATP载体蛋白（AAC）位于线粒体内膜，在线粒体能量代谢中起重要作用。ATP通过AAC从基质转运到细胞质，与此同时ADP逆向转运。克隆的心肌分别来自Bio14.6仓鼠、本实验室培育的J-2-N型心肌病仓鼠和Golden仓鼠。与金仓鼠相比，J-2-N仓鼠的AAC竞赛明显降低。用PCR方法从人成纤维细胞cDNA文库中克隆出AAC的cDNA。然后用人类cDNA筛选金仓鼠AAC cDNA，作为后续步骤的探针。从各品系仓鼠心肌中提取基因组DNA，构建DNA文库。从这些文库中，我们成功地获得了仓鼠心脏AAC的基因组DNA。同时进行RNA斑点杂交分析。AAC mRNA水平以金仓鼠最高，其次是血清CK高的J-2-N、血清CK低的J-2-N和Bio14.6。上述结果提示，AAC mRNA水平的降低可能是J-2-N型心肌病仓鼠心脏代谢异常的机制之一。

项目成果

KATO M: "心筋症ハムスターにおける冠微小循環" Coronary. 10. 107-113 (1993)

加藤 M：“心肌病仓鼠的冠状动脉微循环”冠状动脉。

KATO M: "Effects of Angiotenis-Converting-Enzyme inhibitors and prostagrandin E_1 derivatives on cardiomyopathic hamster." Raben Press New York.The Cardiomyopathic Heart. 157-164 (1994)

KATO M：“血管紧张素转换酶抑制剂和前列腺素 E_1 衍生物对心肌病仓鼠的影响。”

Kato, M: "Effects of Angiotensin-Converting-Enzyme inhibitors and prostagrandin E1 derivatives on cardiomyopathic hamster." The Cardiomyophic Heart, Raven Press. 157-164 (1994)

Kato, M：“血管紧张素转换酶抑制剂和前列腺素 E1 衍生物对心肌病仓鼠的影响。”

KATO M: "Abnormalities of ADP/ATP carrier protein in J-2-N cardiomyopathic hamsters." Mol Cell Biochem.119. 89-94 (1993)

KATO M：“J-2-N 心肌病仓鼠中 ADP/ATP 载体蛋白的异常。”

KATO M: "ADP/ATP carrier protein in cardiomyopachic hamster and the changes of mitochondrial enzyme activity in swimming training rats." 体力研究. 83. 36-44 (1993)

加藤 M：“心肌肥厚仓鼠中的 ADP/ATP 载体蛋白和游泳训练大鼠线粒体酶活性的变化。” 83. 36-44 (1993)。