Although it has been recognized that energy metabolism and mitochondrial structure and functional activity in the immature brain differs from that of the adult, few studies have examined mitochondria specifically at the neuronal synapse during postnatal brain development. In this study, we examined the presynaptic mitochondrial proteome in mice at postnatal day 7 and 42, a period that involves the formation and maturation of synapses. Application of two independent quantitative proteomics approaches – SWATH‐MS and super‐SILAC – revealed a total of 40 proteins as significantly differentially expressed in the presynaptic mitochondria. In addition to elevated levels of proteins known to be involved in ATP metabolic processes, our results identified increased levels of mitoNEET (Cisd1), an iron‐sulfur containing protein that regulates mitochondrial bioenergetics. We found that mitoNEET overexpression plays a cell‐type specific role in ATP synthesis and in neuronal cells promotes ATP generation. The elevated ATP levels in SH‐SY5Y neuroblastoma cells were associated with increased mitochondrial membrane potential and a fragmented mitochondrial network, further supporting a role for mitoNEET as a key regulator of mitochondrial function.

中文翻译：

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突触前线粒体的定量蛋白质组学揭示了产后大脑发育中神经元MitoNEET的过表达和生物学相关性。

尽管已经认识到，未成熟大脑的能量代谢，线粒体结构和功能活性与成年人不同，但是很少有研究专门针对产后大脑发育过程中神经元突触的线粒体进行研究。在这项研究中，我们在出生后第7天和第42天检查了小鼠突触前线粒体蛋白质组，这一时期涉及突触的形成和成熟。两种独立的定量蛋白质组学方法（SWATH-MS和super-SILAC）的应用揭示了总共40种在突触前线粒体中差异显着表达的蛋白质。除了已知与ATP代谢过程有关的蛋白质水平升高外，我们的结果还发现mitoNEET（Cisd1）的水平升高，一种含铁硫的蛋白质，可调节线粒体的生物能。我们发现mitoNEET的过表达在ATP合成中起细胞类型的特定作用，而在神经元细胞中则促进ATP的产生。SH‐SY5Y神经母细胞瘤细胞中的ATP水平升高与线粒体膜电位增加和线粒体网络碎片化有关，进一步支持了mitoNEET作为线粒体功能的关键调节剂的作用。