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. Author manuscript; available in PMC: 2016 Jan 7.
Published in final edited form as: Neuron. 2014 Dec 18;85(1):101–115. doi: 10.1016/j.neuron.2014.11.018

Figure 2. Astroglial C3 alters synaptic and dendritic morphology.

Figure 2

(A) Double-immunostaining of wild-type or IκBα KO astroglia (WTA or KOA) co-cultured neurons with anti-synaptophysin (Syn) and anti-MAP2 (MAP2) antibodies. (B) Same as (A) except that anti-VGluT1 (VGluT1) antibody was used instead of Syn. Images underneath each panel are enlarged view of the bracketed areas. (C) Quantification of number of Syn+MAP2+, VGluT1+MAP2+ or VGAT+MAP2+ synaptic puncta per 10 μm of dendrite in WTA and KOA co-cultured neurons. NWTA Syn=31; NKOA Syn=28; NWTA VGluT1=57; NKOA VGluT1=52; NWTA VGAT=45; NKOA VGAT=52. (D) Quantification of total MAP2-positive dendritic length in WTA and KOA co-cultured neurons. NWTA=38; NKOA=42. (E) Representative dendritic structure, (F) Quantification of dendritic complexity of WTA and KOA co-cultured neurons by Sholl analysis. NWTA=39; NKOA=44. (G) Double staining of Syn and MAP2 of WT neurons treated with vehicle (PBS) or 5 μg/ml C3. (H) Quantified synaptic density of neurons treated with PBS or C3 at 1, 2, or 5 μg/ml. NPBS=36; N1μg/ml=33; N2μg/ml=37; N5μg/ml=29. (I) Representative dendritic structures of WT neurons treated with PBS or 5 μg/ml C3. (J) Dendritic complexity quantification of WT neurons treated with PBS or C3 at 1, 2, and 5 μg/ml. NPBS=116; N1μg/ml=66; N2μg/ml=53; N5μg/ml=68. Scale bar: 10 μm in (A, B, G) and 20 μm in (E, I). C and D: Student’s t-test. F: Two-way ANOVA. H: One-way ANOVA followed by Bonferroni post-hoc analysis. J: Two-way ANOVA followed by Bonferroni post-hoc analysis. *P < 0.05; **P < 0.01; ***P < 0.001; NS: non-significant. See also Figure S2.