Fig. 8

Wild Type (WT) astrocytes ameliorate morphological defects in Ppt1 deficient (Ppt1^−/−) neurons. a WT and Ppt1^−/− astrocytes and neurons were cultured together for 2 or 7 days, and stained with MAP2 (green) and cleaved caspase 3 (CC3, red) to examine cell survival and neuronal morphology. b After both 2 and 7 days in culture, the percentage of CC3 expressing cells was significantly higher in both WT and Ppt1^−/− co-cultures when grown with Ppt1^−/− astrocytes. c After both 2 and 7 days in culture, soma size in all Ppt1^−/− culture conditions was significantly smaller than in WT monocultures, and WT neuron/WT astrocyte co-cultures. Although Ppt1^−/− astrocytes had little effect upon Ppt1^−/− neuronal soma size, WT neuron soma size was significantly reduced when grown with Ppt1−/−astrocytes after 2 and 7 days in culture. d After 2 days in co-culture, the mean neurite length was shorter in Ppt1^−/− neurons under all conditions. Following 7 days in co-culture, Ppt1^−/− astrocytes had a detrimental impact on both WT and Ppt1^−/− neurite outgrowth, whereas WT astrocytes improved neurite outgrowth in Ppt1^−/− neurons. e Axon length in all Ppt1^−/− cultures was significantly shorter than in WT neuron and WT neuron/WT astrocyte co-cultures after 2 days. After 7 days, Ppt1^−/− axon length was significantly reduced in the presence of Ppt1^−/− astrocytes, whereas WT astrocytes significantly improved axon growth. f The number of primary neurites was significantly lower in in all Ppt1^−/− cultures as well as WT neuron/ Ppt1^−/− astrocyte cultures. g Secondary neurite number was significantly reduced in WT neuron/ Ppt1^−/− astrocyte cultures, Ppt1^−/− neuron cultures Ppt1^−/− neuron/WT astrocyte and Ppt1^−/− neuron/ Ppt1^−/− astrocyte co-cultures compared to WT neuron cultures and WT neuron/WT astrocyte cultures. Secondary neurite number remained significantly lower in Ppt1^−/− neuron/ Ppt1^−/− astrocyte cultures than in WT neuron/ Ppt1^−/− astrocyte cultures, Ppt1^−/− astrocyte cultures and Ppt1^−/− neuron/ Ppt1^−/− astrocyte cultures. h The number of tertiary neurites was significantly lower in WT neuron/ Ppt1^−/− astrocyte cultures, Ppt1^−/− neuron cultures, Ppt1^−/− neuron/WT astrocyte and Ppt1^−/− neuron/ Ppt1^−/−astrocyte co cultures than in WT neuronal cultures and WT neuron/WT astrocyte cultures. (Data shown as Mean ± SEM using a one-way ANOVA, n = 3, # represents significant difference to WT neuron cultures, + represents significant difference to WT neuron/WT astrocyte cultures)