突变对过氧化物酶体结构或脂质代谢没有明显影响。
Mutation Has No Apparent Effects on Peroxisome Structure or Lipid Metabolism.
机构信息
Kidney Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland.
Clinical Mass Spectrometry Core, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland.
出版信息
Kidney360. 2021 Jul 16;2(10):1576-1591. doi: 10.34067/KID.0000962021. eCollection 2021 Oct 28.
BACKGROUND
Multiple studies of tissue and cell samples from patients and preclinical models of autosomal dominant polycystic kidney disease report abnormal mitochondrial function and morphology and suggest metabolic reprogramming is an intrinsic feature of this disease. Peroxisomes interact with mitochondria physically and functionally, and congenital peroxisome biogenesis disorders can cause various phenotypes, including mitochondrial defects, metabolic abnormalities, and renal cysts. We hypothesized that a peroxisomal defect might contribute to the metabolic and mitochondrial impairments observed in autosomal dominant polycystic kidney disease.
METHODS
Using control and kidney epithelial cells, we investigated peroxisome abundance, biogenesis, and morphology by immunoblotting, immunofluorescence, and live cell imaging of peroxisome-related proteins and assayed peroxisomal specific -oxidation. We further analyzed fatty acid composition by mass spectrometry in kidneys of mice. We also evaluated peroxisome lipid metabolism in published metabolomics datasets of mutant cells and kidneys. Lastly, we investigated if the C terminus or full-length polycystin-1 colocalize with peroxisome markers by imaging studies.
RESULTS
Peroxisome abundance, morphology, and peroxisome-related protein expression in cells were normal, suggesting preserved peroxisome biogenesis. Peroxisomal -oxidation was not impaired in cells, and there was no obvious accumulation of very-long-chain fatty acids in kidneys of mutant mice. Reanalysis of published datasets provide little evidence of peroxisomal abnormalities in independent sets of mutant cells and cystic kidneys, and provide further evidence of mitochondrial fatty acid oxidation defects. Imaging studies with either full-length polycystin-1 or its C terminus, a fragment previously shown to go to the mitochondria, showed minimal colocalization with peroxisome markers restricted to putative mitochondrion-peroxisome contact sites.
CONCLUSIONS
Our studies showed that loss of does not disrupt peroxisome biogenesis nor peroxisome-dependent fatty acid metabolism.
背景
多项针对常染色体显性多囊肾病患者的组织和细胞样本以及临床前模型的研究报告称，这些样本中的线粒体功能和形态异常，并表明代谢重编程是该疾病的固有特征。过氧化物酶体在物理和功能上与线粒体相互作用，先天性过氧化物酶体生物发生障碍可导致多种表型，包括线粒体缺陷、代谢异常和肾脏囊肿。我们假设过氧化物酶体缺陷可能导致常染色体显性多囊肾病中观察到的代谢和线粒体损伤。
方法
我们使用对照和 肾脏上皮细胞，通过免疫印迹、过氧化物酶体相关蛋白的免疫荧光和活细胞成像以及过氧化物酶体特异性 -氧化测定来研究过氧化物酶体的丰度、生物发生和形态。我们进一步通过质谱分析了 小鼠肾脏中的脂肪酸组成。我们还评估了发表的 突变细胞和肾脏代谢组学数据集的过氧化物酶体脂质代谢。最后，我们通过成像研究研究了 C 端或全长多囊蛋白-1是否与过氧化物酶体标志物共定位。
结果
细胞中的过氧化物酶体丰度、形态和过氧化物酶体相关蛋白表达正常，表明过氧化物酶体生物发生得到保留。 细胞中的过氧化物体 -氧化未受损，突变小鼠肾脏中也没有明显的极长链脂肪酸积累。对发表数据集的重新分析提供了很少有证据表明在独立的 突变细胞和囊性肾脏组中存在过氧化物体异常，并进一步证明了线粒体脂肪酸氧化缺陷。使用全长多囊蛋白-1或其 C 端（先前显示进入线粒体的片段）的成像研究显示，与过氧化物酶体标志物的最小共定位仅限于假定的线粒体-过氧化物酶体接触位点。
结论
我们的研究表明， 的缺失不会破坏过氧化物酶体的生物发生或过氧化物酶体依赖的脂肪酸代谢。
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