Fig. 7.

In vivo bioluminescent imaging indicates AST-004 increases ATP in astrocytes. a Photon flux plots of TBI-injured transgenic mice expressing the Luciferase-reporter gene in astrocytes (GFAP promoter, Jackson Labs) after intraperitoneal injections (IP) of synthetic D-luciferin analog Cycluc1 (5 mM, 100 µl). Bioluminescent signals were recorded with a Xenogen IVIS Spectrum Imager, beginning 5 min after Cycluc1 injection (green arrow). Mice were then injected IP with either vehicle or AST-004 at 4 min (blue arrow); the photon flux was measured at the time of injection and then compared to the photon flux level at 20 min (red arrow). Every 24 h after the initial flux recordings, the same mice were injected again for another photon flux recording, alternating the identity of the test injection between vehicle and AST-004. b Histogram plot showing photon flux differences at 20 min compared to corresponding levels at 4 min. Note mice injected with AST-004 exhibited positive photon flux difference compared to a loss of photon flux (negative) in vehicle-treated mice, consistent with higher ATP production in astrocytes of AST-004-treated mice. Number of mice used for TBI + vehicle: 2 females, for TBI + AST-004: 2 females. In total, 12 measurements were used from these mice every 24 h. alternating between vehicle and AST-004