Characteristics of calcium signaling in astrocytes induced by photostimulation with femtosecond laser

摘要

Astrocytes have been identified to actively contribute to brain functions through Ca~(2+) signaling, serving as a bridge to communicate with neurons and other brain cells. However, conventional stimulation techniques are hard to apply to delicate investigations on astrocytes. Our group previously reported photostimulation with a femtosecond laser to evoke astrocytic calcium (Ca~(2+)) waves, providing a noninvasive and efficient approach with highly precise targeting. In this work, detailed characteristics of astrocytic Ca~(2+) signaling induced by photostimulation are presented. In a purified astrocytic culture, after the illumination of a femtosecond laser onto one cell, a Ca~(2+) wave throughout the network with reduced speed is induced, and intracellular Ca~(2+) oscillations are observed. The intercellular propagation is pharmacologically confirmed to be mainly mediated by ATP through P_2Y receptors. Different patterns of Ca~(2+) elevations with increased amplitude in the stimulated astrocyte are discovered by varying the femtosecond laser power, which is correspondingly followed by broader intercellular waves. These indicate that the strength of photogenerated Ca~(2+) signaling in astrocytes has a positive relationship with the stimulating laser power. Therefore, distinct Ca~(2+) signaling is feasibly available for specific studies on astrocytes by employing precisely controlled photostimulation.