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Figure 5


Fig. 5. AQP4 knockout alters spontaneous Ca2+ oscillations and suppresses high concentration KCl-induced Ca2+ transient in ANSCs. (A) Double immunocytochemistry of nestin (green) and AQP4 (red) in cells migrated out of adherent neurospheres 24 hours after plating. (B) Spontaneous Ca2+ oscillations recorded from three Aqp4+/+ and Aqp4–/– ANSCs (the oscillations of each cell are represented by a different color) loaded with fluo-3 AM. (C) Statistical analysis of data obtained from three independent experiments indicated that the mean amplitude ({Delta}F/F0) of spontaneous Ca2+ oscillations in Aqp4–/– ANSCs was significantly lower than that of Aqp4+/+ control (*P<0.01). The frequency of spontaneous Ca2+ oscillations recorded from Aqp4–/– ANSCs was significantly enhanced compared with that of Aqp4+/+ control (*P<0.01). (D) Sequence of images recorded over 90 seconds from Aqp4+/+ and Aqp4–/– ANSCs loaded with fluo-3 AM following KCl-induced depolarization. (E) Fluo-3 fluorescence expressed as {Delta}F/F0; increased fluorescence indicates elevated [Ca2+]i. There was a significant inhibition of peak amplitudes and no significant sustained phase durations in Aqp4–/– ANSCs compared with Aqp4+/+ control. (F) Peak amplitudes of the KCl-induced Ca2+ transient show a significant decrease in Aqp4–/– ANSCs compared with that in the Aqp4+/+ control (*P<0.001). Scale bars: 50 µm.





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