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. 2019 Dec 12;8:e50712. doi: 10.7554/eLife.50712

Figure 5. Altered distribution of GABAAR clustering in InSyn1 KO neurons.

(A) Representative images of WT and InSyn1 KO neurons at DIV13, immunostained with α1, α2, β3, and y2 GABAAR subunits (green), and Vgat (magenta). Scale bars, 10 µm. (B) Bar graphs showing the quantification of normalized cluster density and area size of each GABAAR and Vgat from perisomatic or neurite regions. GABAARα1 density (Two-tailed t test, Perisomatic; WT n = 13, KO n = 16, p=0.7421. Neurite; WT n = 14, KO n = 17, p=0.1589), and area (Two-tailed t test, Perisomatic; WT n = 14, KO n = 17, p=0.4237. Neurite; WT n = 12, KO n = 16, p=0.4012). GABAARα2 density (Two-tailed t test, Perisomatic; WT n = 16, KO n = 15, p=0.0179. Neurite; WT n = 17, KO n = 18, p=0.0021), and area size (Two-tailed t test, Perisomatic; WT n = 16, KO n = 16, p=0.1083. Neurite; WT n = 16, KO n = 16, p=0.0009). GABAARβ3 density (Two-tailed t test, Perisomatic; WT n = 19, KO n = 16, p=0.1874. Neurite; WT n = 17, KO n = 18, p=0.0522), and area size (Two-tailed t test, Perisomatic; WT n = 20, KO n = 15, p=0.8574. Neurite; WT n = 19, KO n = 15, p=0.0353). GABAARγ2 density (Two-tailed t test, Perisomatic; WT n = 14, KO n = 10, p=0.0254. Neurite; WT n = 16, KO n = 12, p=0.2996), and area size (Two-tailed t test, Perisomatic; WT n = 13, KO n = 16, p=0.3262. Neurite; WT n = 12, KO n = 15, p=0.1271). Of note, no significant difference was found in Vgat cluster quantifications (Two-tailed t-test. GABAARα1, Perisomatic; WT n = 13, KO n = 17, p=0.5510. Neurite; WT n = 13, KO n = 15, p=0.2996. GABAARα2, Perisomatic; WT n = 17, KO n = 16, p=0.8858. Neurite; WT n = 17, KO n = 17, p=0.5657. GABAARβ3, Perisomatic; WT n = 19, KO n = 16, p=0.3011. Neurite; WT n = 19, KO n = 16, p=0.5536. GABAARγ2, Perisomatic; WT n = 15, KO n = 12, p=0.9152. Neurite; WT n = 17, KO n = 15, p=0.7124). *p<0.05, ***p<0.001.

Figure 5.

Figure 5—figure supplement 1. Quantification of GABAARs in αDG-negative cells.

Figure 5—figure supplement 1.

(A) Representative images of WT and InSyn1 KO hippocampal neurons at DIV13, labeled with antibodies to αDG (red), GABAARs (green), and Tuj-1 (blue). αDG positive (arrows) and negative (arrowheads) are indicated. Scale bars, 10 µm. (B) Bar graphs showing normalized GABAARs cluster density or area size either at perisomatic or at neurite regions from αDG negative cells. GABAARα1 perisomatic cluster density (WT n = 19, KO n = 20, two-tailed t-test, p=0.0185), neurite cluster density (WT n = 20, KO n = 19, two-tailed t-test, p=0.0640), perisomatic cluster area (WT n = 19, KO n = 20, two-tailed t-test, p=0.1026), and neurite cluster area (WT n = 19, KO n = 19, two-tailed t-test, p=0.3932). GABAARα2 perisomatic cluster density (WT n = 16, KO n = 12, two-tailed t-test, p=0.3015), neurite cluster density (WT n = 16, KO n = 14, two-tailed t-test, p=0.0039), perisomatic cluster area (WT n = 17, KO n = 12, two-tailed t-test, p=0.3511), and neurite cluster area (WT n = 16, KO n = 12, two-tailed t-test, p=0.7319). *p<0.05, **p<0.001. Scale bars, 10 um.