Photochemically Initiated Intracellular Astrocytic Calcium Waves in Living Mice Using Two-Photon Uncaging of IP3

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Abstract
We have developed a caged IP3 analogue for two-photon photolysis in living animals. This probe is a cell permeable version and was coloaded with a fluorescent Ca2+ dye into astrocytes in layer 1 of the somatosensory cortex of anesthetized mice. Two-photon irradiation of single cells at 720 nm produced rapid and robust increases in intracellular Ca2+ concentrations monitored using two-photon microscopy at 950 nm. The photoevoked intracellular Ca2+ waves were similar in magnitude to intrinsic signals in wild type mice. These waves did not propagate to other cells beyond the targeted astrocyte. In contrast, we observed intercellular astrocytic Ca2+ waves in two mouse models of familial Alzheimer’s disease. These data suggest that Alzheimer’s might perturb gliotransmission but not IP3 signaling per se in mouse models of the disease.

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