An unexpected effect of risperidone reveals a nonlinear relationship between cytosolic Ca²⁺ and mitochondrial Ca²⁺...

Mitochondria actively contribute to cellular Ca²⁺ homeostasis. The molecular mechanisms of mitochondrial Ca²⁺ uptake and release are well characterized and are attributed to the multi-protein assembly of the mitochondrial Ca²⁺ uniporter complex (MCUC) and the mitochondrial sodium-calcium exchanger (NCLX), respectively. Hence, Ca²⁺ transfer from the endoplasmic reticulum (ER) and store-operated Ca²⁺ entry (SOCE) into the mitochondrial matrix has been quantitatively visualized on the subcellular level using targeted fluorescent biosensors. However, a correlation between the amplitude of cytosolic Ca²⁺ elevation with that in the mitochondrial matrix has not been investigated in detail so far. In the present study, we combined the Ca²⁺-mobilizing agonist histamine with the H₁-receptor antagonist risperidone to establish a well-tunable experimental approach allowing the correlation between low, slow, high, and fast cytosolic and mitochondrial Ca²⁺ signals in response to inositol 1,4,5-trisphosphate (IP₃)-triggered ER Ca²⁺ release. Our present data confirm a defined threshold in cytosolic Ca²⁺, which is necessary for the activation of mitochondrial Ca²⁺ uptake. Moreover, our data support the hypothesis of different modes of mitochondrial Ca²⁺ uptake depending on the source of the ion (i.e., ER vs SOCE).