Analyzing Mitochondrial Calcium Influx in Isolated Mitochondria
Analyzing Mitochondrial Calcium Influx in Isolated Mitochondria

Analyzing Mitochondrial Calcium Influx in Isolated Mitochondria

Methods Mol Biol. 2025;2861:155-164. doi: 10.1007/978-1-0716-4164-4_12.

ABSTRACT

Mitochondria play a crucial role in Ca2+ signaling and homeostasis and can contribute to shaping the cytosolic Ca2+ landscape as well as regulate a variety of pathways including energy production and cell death. Dysregulation of mitochondrial Ca2+ homeostasis promotes pathologies including neurodegenerative diseases, cardiovascular disorders, and metabolic syndromes. The significance of mitochondria to Ca2+ signaling and regulation underscores the value of methods to assess mitochondrial Ca2+ import. Here we present a plate reader-based method using the Ca2+-sensitive fluorescent probe calcium green-5 N to measure mitochondrial Ca2+ import in isolated cardiac mitochondria. This technique can be expanded to measure Ca2+ uptake in mitochondria isolated from other tissue types and from cultured cells.

PMID:39395104 | DOI:10.1007/978-1-0716-4164-4_12