Fig. 2

In vitro mibefradil treatment ameliorates aberrant dendritic morphology of β-III^−/− Purkinje cells. (a) Dissociated control (+/+) and β-III^−/− cerebellar cultures, maintained for 21 days in vitro, immunostained for IP₃R1, a Purkinje cell specific marker. Representative image shows excessive “feathery” dendritic morphology of β-III^−/− Purkinje cells. Scale bar, 50 μm. Lower panels show higher magnification of boxed area. Scale bar, 25 μm. (b) Quantification of number of Purkinje cell dendritic branches in 12-day-old dissociated control (+/+; blue) and β-III^−/− (yellow) cerebellar cultures, in the absence or presence of mibefradil (N = 3 for both genotypes; n = 8–17 for each condition). Number of branches per Purkinje cell significantly reduced for β-III^−/− cultures (yellow) treated with 2 µM mibefradil compared to 0 µM mibefradil, denoted by ##. Statistical differences between control (blue) and β-III^−/− (yellow) cultures denoted by asterixis (*). (c) Quantification of total Purkinje cell dendritic length in 12-day-old dissociated control (+/+; blue) and β-III^−/− (yellow) cerebellar cultures, in the absence or presence of mibefradil (N = 3 for both genotypes; n = 8–17 for each condition). Total dendritic length per Purkinje cell significantly reduced for β-III^−/− (yellow) cultures treated with 2 µM mibefradil compared to 0 µM mibefradil, denoted by #. Statistical differences between control (blue) and β-III^−/− (yellow) cultures denoted by asterixis (*). All data plotted as median, quartiles (25th and 75th percentiles), minimum and maximum with mean indicated by cross. (d) Representative images of β-III^−/− Purkinje cells cultured for 12-days in the absence or presence of 2 µM mibefradil, with latter showing dendritic morphology similar to mibefradil treated control (+/+) Purkinje cell. Scale bar, 25 μm.