Microscopic heat pulse-induced calcium dynamics in single WI-38 fibroblasts

Abstract

Temperature-sensitive Ca²⁺ dynamics occur primarily through transient receptor potential channels, but also by means of Ca²⁺ channels and pumps on the endoplasmic reticulum membrane. As such, cytoplasmic Ca²⁺ concentration ([Ca²⁺]_cyt) is re-equilibrated by changes in ambient temperature. The present study investigated the effects of heat pulses (heating duration: 2 s or 150 s) on [Ca²⁺]_cyt in single WI-38 fibroblasts, which are considered as normal cells. We found that Ca²⁺ burst occurred immediately after short (2 s) heat pulse, which is similar to our previous report on HeLa cells, but with less thermosensitivity. The heat pulses originated from a focused 1455-nm infrared laser light were applied in the vicinity of cells under the optical microscope. Ca²⁺ bursts induced by the heat pulse were suppressed by treating cells with inhibitors for sarco/endoplasmic reticulum Ca²⁺ ATPase (SERCA) or inositol trisphosphate receptor (IP₃R). Long (150 s) heat pulses also induced Ca²⁺ bursts after the onset of heating and immediately after re-cooling. Cells were more thermosensitive at physiological (37°C) than at room (25°C) temperature; however, at 37°C, cells were responsive at a higher temperature (ambient temperature+heat pulse). These results strongly suggest that the heat pulse-induced Ca²⁺ burst is caused by a transient imbalance in Ca²⁺ flow between SERCA and IP₃R, and offer a potential new method for thermally controlling Ca²⁺-regulated cellular functions.