Overexpression of Mitochondrial Calcium Uniporter Causes Neuronal Death

Abstract

Neurodegenerative diseases are a large and heterogeneous group of disorders characterized by selective and progressive death of specific neuronal subtypes. In most of the cases, the pathophysiology is still poorly understood, although a number of hypotheses have been proposed. Among these, dysregulation of Ca²⁺ homeostasis and mitochondrial dysfunction represent two broadly recognized early events associated with neurodegeneration. However, a direct link between these two hypotheses can be drawn. Mitochondria actively participate to global Ca²⁺ signaling, and increases of [Ca²⁺] inside organelle matrix are known to sustain energy production to modulate apoptosis and remodel cytosolic Ca²⁺ waves. Most importantly, while mitochondrial Ca²⁺ overload has been proposed as the no-return signal, triggering apoptotic or necrotic neuronal death, until now direct evidences supporting this hypothesis, especially <i>in vivo</i>, are limited. Here, we took advantage of the identification of the mitochondrial Ca²⁺ uniporter (MCU) and tested whether mitochondrial Ca²⁺ signaling controls neuronal cell fate. We overexpressed MCU both <i>in vitro</i>, in mouse primary cortical neurons, and <i>in vivo</i>, through stereotaxic injection of MCU-coding adenoviral particles in the brain cortex. We first measured mitochondrial Ca²⁺ uptake using quantitative genetically encoded Ca²⁺ probes, and we observed that the overexpression of MCU causes a dramatic increase of mitochondrial Ca²⁺ uptake both at resting and after membrane depolarization. MCU-mediated mitochondrial Ca²⁺ overload causes alteration of organelle morphology and dysregulation of global Ca²⁺ homeostasis. Most importantly, MCU overexpression <i>in vivo</i> is sufficient to trigger gliosis and neuronal loss. Overall, we demonstrated that mitochondrial Ca²⁺ overload is <i>per se</i> sufficient to cause neuronal cell death both <i>in vitro</i> and <i>in vivo</i>, thus highlighting a potential key step in neurodegeneration.