Journal: The Journal of Neuroscience
Article Title: Cytoskeletal Assembly and ATP Release Regulate Astrocytic Calcium Signaling
Figure Lengend Snippet: Calcium wave activity is suppressed by cytochalasin D in established astrocytic cultures. Confluent astrocytic cultures were loaded with fluo-3, and calcium waves were evoked by mechanical stimulation. Upper row , A control untreated culture propagates a calcium wave that includes > 25 cells. Middle row , Astrocytes pretreated with cytochalasin D for 5 min only propagate calcium increases to a few astrocytes. Lower row , Culture treated with cytochalasin D for 10 min was not capable of propagating a calcium wave, despite a robust calcium increment in the stimulated cell. Actin fibers were visualized in the same field ( right , in red ) by fixing and staining with Texas Red–phalloidin immediately after analysis of calcium signaling. Note the gradual disappearance of stress fibers in cytochalasin D-treated cultures. Fluorescence intensities of the fluo-3 probe corresponding to relative calcium levels were monitored 1, 7, 15, and 23 sec after stimulation by confocal microscopy. In all frames, background counts were subtracted. Scale bar, 10 μm.
Article Snippet: After fluo-3 resting signal was recorded, 25 μ m ATP or 25 μ m bradykinin (Sigma) was applied.
Techniques: Activity Assay, Staining, Fluorescence, Size-exclusion Chromatography, Confocal Microscopy