Yao Pamela J, Munk Rachel, Gorospe Myriam, Kapogiannis Dimitrios
Laboratory of Clinical Investigation, National Institute on Aging Intramural Research Program, National Institutes of Health, Baltimore, MD, USA.
Laboratory of Genetics and Genomics, National Institute on Aging Intramural Research Program, National Institutes of Health, Baltimore, MD, USA.
Heliyon. 2023 Feb 22;9(3):e13888. doi: 10.1016/j.heliyon.2023.e13888. eCollection 2023 Mar.
Studying mitochondrial respiration capacity is essential for gaining insights into mitochondrial functions. In frozen tissue samples, however, our ability to study mitochondrial respiration is restricted by damage elicited to the inner mitochondrial membranes by freeze-thaw cycles. We developed an approach that combines multiple assays and is tailored towards assessing mitochondrial electron transport chain and ATP synthase in frozen tissues. Using small amounts of frozen tissue, we systematically analyzed the quantity as well as activity of both the electron transport chain complexes and ATP synthase in rat brains during postnatal development. We reveal a previously little-known pattern of increasing mitochondrial respiration capacity with brain development. In addition to providing proof-of-principle evidence that mitochondrial activity changes during brain development, our study details an approach that can be applicable to many other types of frozen cell or tissue samples.
研究线粒体呼吸能力对于深入了解线粒体功能至关重要。然而，在冷冻组织样本中，我们研究线粒体呼吸的能力受到冻融循环对内线粒体膜造成的损伤的限制。我们开发了一种方法，该方法结合了多种检测手段，专门用于评估冷冻组织中的线粒体电子传递链和ATP合酶。我们使用少量冷冻组织，系统地分析了大鼠大脑在出生后发育过程中电子传递链复合物和ATP合酶的数量及活性。我们揭示了一种此前鲜为人知的随着大脑发育线粒体呼吸能力增强的模式。除了提供线粒体活性在大脑发育过程中发生变化的原理性证据外，我们的研究还详细介绍了一种可应用于许多其他类型冷冻细胞或组织样本的方法。
