From INSERM, UMR-S1028.
J Biol Chem. 2014 Jan 24;289(4):2261-76. doi: 10.1074/jbc.M113.490862. Epub 2013 Dec 9.
Degradation of damaged mitochondria by mitophagy is an essential process to ensure cell homeostasis. Because neurons, which have a high energy demand, are particularly dependent on the mitochondrial dynamics, mitophagy represents a key mechanism to ensure correct neuronal function. Collapsin response mediator proteins 5 (CRMP5) belongs to a family of cytosolic proteins involved in axon guidance and neurite outgrowth signaling during neural development. CRMP5, which is highly expressed during brain development, plays an important role in the regulation of neuronal polarity by inhibiting dendrite outgrowth at early developmental stages. Here, we demonstrated that CRMP5 was present in vivo in brain mitochondria and is targeted to the inner mitochondrial membrane. The mitochondrial localization of CRMP5 induced mitophagy. CRMP5 overexpression triggered a drastic change in mitochondrial morphology, increased the number of lysosomes and double membrane vesicles termed autophagosomes, and enhanced the occurrence of microtubule-associated protein 1 light chain 3 (LC3) at the mitochondrial level. Moreover, the lipidated form of LC3, LC3-II, which triggers autophagy by insertion into autophagosomes, enhanced mitophagy initiation. Lysosomal marker translocates at the mitochondrial level, suggesting autophagosome-lysosome fusion, and induced the reduction of mitochondrial content via lysosomal degradation. We show that during early developmental stages the strong expression of endogenous CRMP5, which inhibits dendrite growth, correlated with a decrease of mitochondrial content. In contrast, the knockdown or a decrease of CRMP5 expression at later stages enhanced mitochondrion numbers in cultured neurons, suggesting that CRMP5 modulated these numbers. Our study elucidates a novel regulatory mechanism that utilizes CRMP5-induced mitophagy to orchestrate proper dendrite outgrowth and neuronal function.
受损线粒体通过自噬作用的降解是确保细胞内稳态的一个必要过程。由于神经元具有较高的能量需求,它们特别依赖于线粒体动力学,因此自噬作用是确保正确神经元功能的关键机制。 collapsin 反应介质蛋白 5(CRMP5)属于细胞溶质蛋白家族,在神经发育过程中参与轴突导向和神经突生长信号传递。CRMP5 在大脑发育过程中高度表达,通过在早期发育阶段抑制树突生长,在调节神经元极性方面发挥重要作用。在这里,我们证明了 CRMP5 体内存在于脑线粒体中,并靶向于线粒体内膜。CRMP5 的线粒体定位诱导自噬作用。CRMP5 的过表达导致线粒体形态发生剧烈变化,增加了溶酶体的数量和双膜囊泡(称为自噬体),并增强了微管相关蛋白 1 轻链 3(LC3)在线粒体水平上的表达。此外,LC3 的脂质化形式 LC3-II 通过插入自噬体触发自噬,增强了自噬的起始。溶酶体标记物在线粒体水平上易位,表明自噬体-溶酶体融合,并通过溶酶体降解诱导线粒体含量减少。我们表明,在早期发育阶段,强烈表达的内源性 CRMP5 抑制树突生长,与线粒体含量减少相关。相比之下,在后期敲低或减少 CRMP5 的表达会增加培养神经元中的线粒体数量,表明 CRMP5 调节了这些数量。我们的研究阐明了一种新的调节机制,利用 CRMP5 诱导的自噬作用来协调适当的树突生长和神经元功能。