Departamento de Química Fundamental, Av. Professor Lineu Prestes, 748, 05508-000, São Paulo, SP, Brazil.
Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua três de Maio, 100, 04044-020, São Paulo, Brazil.
Anal Chim Acta. 2021 Feb 15;1146:88-97. doi: 10.1016/j.aca.2020.12.030. Epub 2020 Dec 29.
Metabolic analysis in animals is usually either evaluated as whole-body measurements or in isolated tissue samples. To reveal tissue specificities in vivo, this study uses scanning electrochemical microscopy (SECM) to provide localized oxygen consumption rates (OCRs) in different regions of single adult Caenorhabditis elegans individuals. This is achieved by measuring the oxygen reduction current at the SECM tip electrode and using a finite element method model of the experiment that defines oxygen concentration and flux at the surface of the organism. SECM mapping measurements uncover a marked heterogeneity of OCR along the worm, with high respiration rates at the reproductive system region. To enable sensitive and quantitative measurements, a self-referencing approach is adopted, whereby the oxygen reduction current at the SECM tip is measured at a selected point on the worm and in bulk solution (calibration). Using genetic and pharmacological approaches, our SECM measurements indicate that viable eggs in the reproductive system are the main contributors in the total oxygen consumption of adult Caenorhabditis elegans. The finding that large regional differences in OCR exist within the animal provides a new understanding of oxygen consumption and metabolic measurements, paving the way for tissue-specific metabolic analyses and toxicity evaluation within single organisms.
动物的代谢分析通常要么作为全身测量值进行评估,要么在分离的组织样本中进行。为了在体内揭示组织特异性,本研究使用扫描电化学显微镜 (SECM) 提供单个成年秀丽隐杆线虫个体不同区域的局部耗氧率 (OCR)。这是通过测量 SECM 尖端电极处的氧气还原电流,并使用实验的有限元方法模型来实现的,该模型定义了生物体表面的氧气浓度和通量。SECM 映射测量揭示了沿蠕虫的 OCR 存在明显的异质性,生殖系统区域的呼吸率很高。为了实现灵敏和定量的测量,采用了自参考方法,其中在蠕虫的选定点和在 bulk solution(校准)处测量 SECM 尖端处的氧气还原电流。通过遗传和药理学方法,我们的 SECM 测量表明,生殖系统中的存活卵子是成年秀丽隐杆线虫总耗氧量的主要贡献者。在动物体内存在 OCR 大的区域性差异的发现为氧耗和代谢测量提供了新的认识,为单个生物体中的组织特异性代谢分析和毒性评估铺平了道路。