Periodic Reporting for period 3 - SynapticMitochondria (Quality Control and Maintenance of Synaptic Mitochondria)
Reporting period: 2019-09-01 to 2021-02-28
How mitochondria sense their dysfunction is unclear. Even more intriguing is the question how they decide whether their failure should lead to removal of the organelle or dismissal of the complete neuron via cell death. We anticipate that these decisions are not only operational during disease, but might constitute a fundamental mechanism relevant for maintenance of synaptic activity and establishment of new synapses.
SYNAPTICMITOCHONDRIA aims to uncover uncharacterized novel components of synaptic mitochondria and to scrutinize quality control mechanisms that synaptic mitochondria use to sense and maintain their intrinsic function in a healthy brain. Even though mitochondrial function has been implicated in synapses maintaining their robustness, little is known about the composition, maintenance and intrinsic properties of synaptic mitochondria. Here we propose to use two complementary approaches to address these questions. A proteomic approach will be used to identify the protein fingerprint of synaptic mitochondria and to compare them to mitochondria from other tissues. Additionally, the identification of key players of the proposed regulatory pathways involved in intrinsic mitochondria quality control will also be attained. In a complimentary approach, we will exploit our findings and use in vitro and in vivo experimental approaches to measure mitochondrial function of synaptic versus non-synaptic mitochondria and the relevance of those changes for synaptic function. Our work will unravel the specific properties of synaptic mitochondria and provide much needed insight in their hypothesized predominant role in neurodegenerative disorders.
The first aim of this research project is to identify and characterize novel components of synaptic mitochondria. For this, we have determined the proteomic profile that discriminates synaptic mitochondria from non-synaptic mitochondria, and through Bioinformatics analysis we have identified our 10 top candidate proteins. At present, we are assessing the protein expression pattern in these two distinct mitochondrial pools, as well as, the role of our candidate proteins in specific mitochondrial regulatory pathways. Additionally, we also are assessing the overall mitochondrial properties of these neuronal mitochondrial pools. For this, we are assessing cristae organization, bionenergetic profile and overall mitochondrial capacity of synaptic and non-synaptic mitochondria.
The second aim of this research project is to pinpoint mitochondrial quality control (MQC) mechanisms that synaptic mitochondria use to sense and maintain their intrinsic function. We have established reporter cell lines that will aid in understanding the cross-talk between PINK1/PARL/PGAM5. Additionally, we also find ourselves well positioned to scrutinize the quality control function of the mitochondrial gatekeeper protein PINK1 in different neural cell types. The understanding of the crosstalk mediated by PINK1 between this different cell types within the context of a healthy brain will determine how these cells go astray in a neurodegenerative scenario.
Our integrative approach, based on using an unbiased approach and a candidate gene approach, is providing a unifying model revealing the mechanisms involved in maintaining a healthy pool of mitochondria at the synapse.
This work will reveal a proteome specific to synaptic mitochondria and unravel the quality control mechanisms that regulate synaptic mitochondria in a healthy and damaged brain. Additionally, we will have developed a unique array of methodology to evaluate the function of mitochondria in synapses which will position us in a unique way to address the many speculative links made between mitochondria and neurodegenerative disorders. Finally, a successful outcome of this research project will enable the identification of molecular mechanisms that cause neurodegenerative disorders, opening the avenues to new therapeutic approaches and moreover, to a contribution of overall brain function.