Regulators of Mitochondrial Transport and Dynamics in Neurons

In addition to being the “powerhouse of the cell,” mitochondria play critical roles in mediating calcium buffering, apoptosis, and necrosis. They are also a major source of reactive oxygen species (ROS), which can have both a signaling role and be damaging to cells. Mitochondria are actively transported within neurons to synapses, and damaged mitochondria – a potential threat to the cell – are transported back to the cell body for removal by mitophagy. Mitochondria are also dynamic, undergoing fusion and fission. Fusion is thought to be a mechanism for boosting mitochondrial output and protecting mitochondrial health, whereas fission is thought to be the first step on the way to mitophagy and the removal of damaged mitochondria. Defects in mitochondrial dynamics have a clear role in Parkinson’s Disease. Defects in mitochondrial transport have a clear role in Alzheimer’s Disease. Thus, an understanding of mitochondrial dynamics and transport is important for our understanding of neurological disorders with mitochondrial etiology, as well as our understanding of aging and age-associated diseases. Mitochondrial dynamics as a field has largely been studied in single celled yeast; thus, little is known about the machinery that conducts mitochondrial fission and fusion in specialized tissues like neurons. We are studying mitochondrial dynamics in C. elegans neurons using a mitochondrially-localized GFP reporter, which makes it easy to visualize individual mitochondria in axons and dendrites of live animals. Using this tool, we performed a forward genetic screen for mutants with defects in mitochondrial transport, dynamics, or mitophagy. We are currently cloning and characterize the underlying genes so as to have a complete understanding of the factors that mediate and regulate mitochondrial biology in neurons. 

We also generated a C. elegans transgenic strain that expresses MitoKeima, new reporter for mitochondria undergoing mitophagy.  MitoKeima has a differential, pH-dependent fluorescence excitation spectra that allows one to discriminate healthy mitochondria in the neutral pH of the cytosol from damaged mitochondria in the low pH environments of autophagosomes, autolysosomes, and lysosomes.  Using this and other mitochondrial reporters, we are now examining how mitochondrial dysfunction contributes to a tau-based genetic model of Alzheimer’s Disease.

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Anoxia-reoxygenation regulates mitochondrial dynamics through the hypoxia response pathway, SKN-1/Nrf, and stomatin-like protein STL-1/SLP-2.

Ghose P, Park EC, Tabakin A, Salazar-Vasquez N, Rongo C. PLoS Genet. 2013;9(12):e1004063. doi: 10.1371/journal.pgen.1004063. Epub 2013 Dec 26. PMID: 24385935

Many aerobic organisms encounter oxygen-deprived environments and thus must have adaptive mechanisms to survive such stress. It is important to understand how mitochondria respond to oxygen deprivation given the critical role they play in using oxygen to generate cellular energy. Here we examine mitochondrial stress response in C. elegans, which adapt to extreme oxygen deprivation (anoxia, less than 0.1% oxygen) by entering into a reversible suspended animation state of locomotory arrest. We show that neuronal mitochondria undergo DRP-1-dependent fission in response to anoxia and undergo refusion upon reoxygenation. The hypoxia response pathway, including EGL-9 and HIF-1, is not required for anoxia-induced fission, but does regulate mitochondrial reconstitution during reoxygenation. Mutants for egl-9 exhibit a rapid refusion of mitochondria and a rapid behavioral recovery from suspended animation during reoxygenation; both phenotypes require HIF-1. Mitochondria are significantly larger in egl-9 mutants after reoxygenation, a phenotype similar to stress-induced mitochondria hyperfusion (SIMH). Anoxia results in mitochondrial oxidative stress, and the oxidative response factor SKN-1/Nrf is required for both rapid mitochondrial refusion and rapid behavioral recovery during reoxygenation. In response to anoxia, SKN-1 promotes the expression of the mitochondrial resident protein Stomatin-like 1 (STL-1), which helps facilitate mitochondrial dynamics following anoxia. Our results suggest the existence of a conserved anoxic stress response involving changes in mitochondrial fission and fusion.


Neurite sprouting and synapse deterioration in the aging Caenorhabditis elegans nervous system.

Toth ML, Melentijevic I, Shah L, Bhatia A, Lu K, Talwar A, Naji H, Ibanez-Ventoso C, Ghose P, Jevince A, Xue J, Herndon LA, Bhanot G, Rongo C, Hall DH, Driscoll M. J Neurosci. 2012 Jun 27;32(26):8778-90. doi: 10.1523/JNEUROSCI.1494-11.2012. PMID: 22745480

Caenorhabditis elegans is a powerful model for analysis of the conserved mechanisms that modulate healthy aging. In the aging nematode nervous system, neuronal death and/or detectable loss of processes are not readily apparent, but because dendrite restructuring and loss of synaptic integrity are hypothesized to contribute to human brain decline and dysfunction, we combined fluorescence microscopy and electron microscopy (EM) to screen at high resolution for nervous system changes. We report two major components of morphological change in the aging C. elegans nervous system: (1) accumulation of novel outgrowths from specific neurons, and (2) physical decline in synaptic integrity. Novel outgrowth phenotypes, including branching from the main dendrite or new growth from somata, appear at a high frequency in some aging neurons, but not all. Mitochondria are often associated with age-associated branch sites. Lowered insulin signaling confers some maintenance of ALM and PLM neuron structural integrity into old age, and both DAF-16/FOXO and heat shock factor transcription factor HSF-1 exert neuroprotective functions. hsf-1 can act cell autonomously in this capacity. EM evaluation in synapse-rich regions reveals a striking decline in synaptic vesicle numbers and a diminution of presynaptic density size. Interestingly, old animals that maintain locomotory prowess exhibit less synaptic decline than same-age decrepit animals, suggesting that synaptic integrity correlates with locomotory healthspan. Our data reveal similarities between the aging C. elegans nervous system and mammalian brain, suggesting conserved neuronal responses to age. Dissection of neuronal aging mechanisms in C. elegans may thus influence the development of brain healthspan-extending therapies.