Gene expression analysis in mitochondria from chagasic mice: alterations in specific metabolic pathways
Gene expression analysis in mitochondria from chagasic mice: alterations in specific metabolic pathways
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DOI：
10.1042/bj20040356
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发表时间：
2004-08-01
影响因子：
4.1
通讯作者：
Papaconstantinou, J
中科院分区：
文献类型：
--
作者：
Garg, N;Gerstner, A;Papaconstantinou, J
心肌肥厚和重构可能与线粒体功能障碍有关。在本研究中，我们使用定制设计的线粒体功能相关基因阵列（mitoarray）表征了克氏锥虫感染和进行性疾病严重程度的心脏代谢反应。由已知的，充分表征的线粒体功能相关的cDNA组成的线粒体阵列与P-32标记的cDNA探针杂交，所述探针在感染和疾病发展的早期、急性和慢性阶段从小鼠心肌产生。线粒体芯片成功地鉴定了T.克鲁兹诱导了与线粒体功能和生物发生相关的基因表达的改变，这通过实时逆转录酶PCR得到了进一步证实。值得注意的是脂肪酸代谢所必需的转录物的上调与感染心脏中丙酮酸脱氢酶复合物的mRNA的抑制相关。我们观察到三羧酸循环酶的mRNA没有统计学显著变化。这些结果表明，脂肪酸代谢补偿了丙酮酸脱氢酶复合物的缺陷，为三羧酸循环提供乙酰辅酶A，并且chagglomerate心脏可能不限于减少的能量（NADH和FADH（2））。通过线粒体阵列以及全球基因组分析观察到呼吸链复合物几个亚基的mRNA水平降低，这表明线粒体氧化磷酸化介导的ATP生成能力有限，这可能是恰格拉病心脏稳态的基础。
Cardiac hypertrophy and remodelling in chagasic disease might be associated with mitochondrial dysfunction. In the present study, we characterized the cardiac metabolic responses to Trypanosoma cruzi infection and progressive disease severity using a custom-designed mitoarray (mitochondrial function-related gene array). Mitoarrays consisting of known, well-characterized mitochondrial function-related cDNAs were hybridized with P-32-labelled cDNA probes generated from the myocardium of mice during immediate early, acute and chronic phases of infection and disease development. The mitoarray successfully identified novel aspects of the T. cruzi-induced alterations in the expression of the genes related to mitochondrial function and biogenesis that were further confirmed by real-time reverse transcriptase-PCRs. Of note is the up-regulation of transcripts essential for fatty acid metabolism associated with repression of the mRNAs for pyruvate dehydrogenase complex in infected hearts. We observed no statistically significant changes in mRNAs for the enzymes of tricarboxylic acid cycle. These results suggest that fatty acid metabolism compensates the pyruvate dehydrogenase complex deficiencies for the supply of acetyl-CoA for a tricarboxylic acid cycle, and chagasic hearts may not be limited in reduced energy (NADH and FADH(2)). The observation of a decrease in mRNA level for several subunits of the respiratory chain complexes by mitoarray as well as global genome analysis suggests a limitation in mitochondrial oxidative phosphorylation-mediated ATP-generation capacity as the probable basis for cardiac homoeostasis in chagasic disease.