Mitochondria play a central role in cell bioenergetics and control multiple aspects of neuronal life and death, including regulation of Ca2+ signaling. By buffering Ca2+ during excitation, and subsequently releasing Ca2+ back to the cytosol, mitochondria shape [Ca2+]i signals and regulate numerous Ca2+-dependent functions in neurons, such as excitability, synaptic plasticity and gene expression. However, excessive load of mitochondria with Ca2+ trigger neurotoxic processes in stroke and in Alzheimer's, Parkinson's and Huntington's diseases. In spite of the importance of neuronal mitochondrial Ca2+ transport, the molecules mediating mitochondrial Ca2+ uptake and release in neurons are not known. This knowledge gap presents a major obstacle in our progress toward understanding and therapeutically exploiting mitochondrial functions. Thus, the first objective of this proposal is to identify the molecular components of mitochondrial Ca2+ uptake in neurons. Mitochondria are highly dynamic organelles that rapidly undergo fission and fusion (MFF), which affects transport of mitochondria, synaptic plasticity and neuronal survival. Notably, mitochondrial fission is an early event in stroke, and fragmented mitochondria are prevalent in Alzheimer's and Huntington's disease. Given the central role of mitochondrial Ca2+ transport in neuronal life and death, it is possible that the effects of MFF on neuronal survival are mediated in part through changes in mitochondrial Ca2+ handling, although this idea has not been tested. Thus, our second objective is to determine how MFF status affects mitochondrial Ca2+ transport and Ca2+ homeostasis in neurons. Based on our preliminary data and published literature, we hypothesize that CCDC109A, CCDC109B and MICU1-3 are essential molecular components of mitochondrial Ca2+ uptake in neurons, and that the MFF process provides important control of CCDC109A and/or CCDC109B activities, mitochondrial Ca2+ transport and Ca2+ homeostasis in neurons exposed to neurotoxic conditions. We will use innovative approaches, including genetically encoded mitochondrial Ca2+ sensors, electron probe X-ray microanalysis and novel genetic mouse strains, to test our hypothesis in three specific aims.
Aim 1 will identify the roles of novel proteins CCDC109A, CCDC109B and MICU1, 2 and 3 in mitochondrial Ca2+ uptake in neurons.
Aim 2 will determine how mitochondrial restructuring regulates mitochondrial Ca2+ transport in neurons and examine specific roles of CCDC109A and CCDC109B phosphorylation in this process.
Aim 3 will examine the function of MFF in maintaining neuronal Ca2+ homeostasis under neurotoxic conditions, such as excessive exposure to glutamate and ischemia. This project will provide insight into the molecular organization of mitochondrial Ca2+ transport in neurons and will establish mechanistic links between mitochondrial dynamics, Ca2+ signaling and neuronal Ca2+ homeostasis. We anticipate that these studies will be transformative because they will identify new molecular and genetic tools for exploring many functions of mitochondrial Ca2+ uptake in neurons and may lead to new therapeutics targeting mitochondrial Ca2+ transport and MFF for treating stroke and neurodegeneration.

Public Health Relevance

Stroke and neurodegenerative disorders such as Alzheimer's and Parkinson's diseases are among the leading causes of death and disabilities in the United States and worldwide. Impairment of mitochondrial function is implicated in neuronal damage in stroke and neurodegenerative disorders. The proposed studies will help to better understand how phosphorylation-dependent restructuring of mitochondria stabilizes Ca2+ homeostasis and mitochondrial function in neurons and provides neuronal protection from toxic conditions. These studies may lead to the development of new therapeutic strategies for neuroprotection in stroke and neurodegenerative disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS087068-01
Application #
8673589
Study Section
Special Emphasis Panel (ZRG1-MDCN-T (91))
Program Officer
Stewart, Randall R
Project Start
2014-03-01
Project End
2018-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$330,313
Indirect Cost
$111,563
Name
University of Iowa
Department
Pharmacology
Type
Schools of Medicine
DUNS #
062761671
City
Iowa City
State
IA
Country
United States
Zip Code
52242