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. Author manuscript; available in PMC: 2012 Dec 20.
Published in final edited form as: J Neurochem. 2010 Apr 16;114(1):271–280. doi: 10.1111/j.1471-4159.2010.06749.x

Fig. 5.

Fig. 5

Effects of U-83836E on Ca++ buffering capacity in mitochondria at 3 h following controlled cortical impact traumatic brain injury in male mice. Mice were administered 3 mg/kg U-83836E i.v. 15 min post-injury and mitochondria were isolated using the Ficoll density-gradient isolation technique. Part (a) shows a quantification of mitochondrial Ca++ buffering capacity across experimental groups. The Ca++ buffering capacity was reduced in vehicle-treated injured group compared to sham (non-injured) group and U-83836E treatment ameliorated this posttraumatic reduction in mitochondrial Ca++ buffering capacity. Mitochondrial Ca++ buffering was evaluated in a thermostatically-controlled, continuously stirred cuvette incubated in a Shimadzu RF-5301 spectrofluorimeter set at 37°C. Levels of extra-mitochondrial calcium were measured using Ca++-sensitive indicator calcium green 5 N (CaG5N), a fluorescent calcium-sensitive indicator. Malate and pyruvate (M/P) and ADP were loaded sequentially over 2 min followed by oligomycin (O) at 3 min. Two minutes after adding oligomycin, calcium (32 nmol/L) infusion started at a rate of 0.5µL/min. Part (b) is a typical trace that shows the differences in mitochondrial Ca++ uptake in sham (non-injured), vehicle-treated injured and U-83836E-treated injured mitochondria. Values = mean ± sd. Statistical differences (one-way anova and Student-Neuman–Keuls post hoc test): *p < 0.01 versus sham, #p < 0.05 versus vehicle, n = 4.