Excessive Astrocyte-Derived Neurotrophin-3 Contributes to the Abnormal Neuronal Dendritic Development in a Mouse Model of Fragile X Syndrome
Figure 3
Expression of FMRP reversed neuronal growth in Fmr1 KO ACM.
Cultured astrocytes at DIV 7 were transfected with FMRP expression vectors for 48 h. A, Confocal images showing the transfection efficiency of FMRP vectors. Nuclei were stained with Hoechst33258 (blue). EGFP-positive astrocytes (green) indicated the successful transfection of FMRP vectors. Scale bar = 50 µm. B, Transfection of FMRP vectors resulted in the expression of FMRP in KO astrocytes, whereas the negative empty vectors did not. n = 6 wells from three independent experiments. **P<0.01 compared with the WT astrocytes; ##P<0.01 compared with the empty control group. C, Neuronal dendritic development was significantly improved in ACM of FMRP vectors transfected KO astrocytes. Scale bar = 50 µm. D, Quantification of neurons with at least two short (<50 µm) dendrites. E, Quantification of the total dendritic length per cell. D–E: the number of neurons in WT ACM: n = 213 neurons, KO + empty ACM: n = 236 neurons, KO + FMRP ACM: n = 185 neurons. Data were from three independent experiments. *P<0.05, **P<0.01 compared with the WT ACM; #P<0.05 compared with the empty vector transfected KO ACM. F, The expressions of MAP2, PSD95, and GluR1 were detected by Western blot. G, Band intensities were quantified as percentage of values from WT ACM-treated neurons. n = 6 wells from three independent experiments. *P<0.05, **P<0.01 compared with the WT ACM; #P<0.05 compared with the empty vector transfected KO ACM.