Stimulus-Dependent Regulation of Nuclear Ca2+ Signaling in Cardiomyocytes: A Role of Neuronal Calcium Sensor-1

In cardiomyocytes, intracellular calcium (Ca²⁺) transients are elicited by electrical and receptor stimulations, leading to muscle contraction and gene expression, respectively. Although such elevations of Ca²⁺levels (Ca²⁺]) also occur in the nucleus, the precise mechanism of nuclear Ca²⁺] regulation during different kinds of stimuli, and its relationship with cytoplasmic Ca²⁺] regulation are not fully understood. To address these issues, we used a new region-specific fluorescent protein-based Ca²⁺ indicator, GECO, together with the conventional probe Fluo-4 AM. We confirmed that nuclear Ca²⁺ transients were elicited by both electrical and receptor stimulations in neonatal mouse ventricular myocytes. Kinetic analysis revealed that electrical stimulation-elicited nuclear Ca²⁺ transients are slower than cytoplasmic Ca²⁺ transients, and chelating cytoplasmic Ca²⁺ abolished nuclear Ca²⁺ transients, suggesting that nuclear Ca²⁺ are mainly derived from the cytoplasm during electrical stimulation. On the other hand, receptor stimulation such as with insulin-like growth factor-1 (IGF-1) preferentially increased nuclear Ca²⁺] compared to cytoplasmic Ca²⁺]. Experiments using inhibitors revealed that electrical and receptor stimulation-elicited Ca²⁺ transients were mainly mediated by ryanodine receptors and inositol 1,4,5-trisphosphate receptors (IP3Rs), respectively, suggesting different mechanisms for the two signals. Furthermore, IGF-1-elicited nuclear Ca²⁺ transient amplitude was significantly lower in myocytes lacking neuronal Ca²⁺ sensor-1 (NCS-1), a Ca²⁺ binding protein implicated in IP₃R-mediated pathway in the heart. Moreover, IGF-1 strengthened the interaction between NCS-1 and IP₃R. These results suggest a novel mechanism for receptor stimulation-induced nuclear Ca²⁺] regulation mediated by IP3R and NCS-1 that may further fine-tune cardiac Ca²⁺ signal regulation.