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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and <t>PSD95-YFP</t> (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.
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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and <t>PSD95-YFP</t> (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.
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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and <t>PSD95-YFP</t> (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.
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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and <t>PSD95-YFP</t> (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.
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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and <t>PSD95-YFP</t> (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.
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(A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and PSD95-YFP (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.

Journal: Cell reports

Article Title: AMIGO2 Scales Dendrite Arbors in the Retina

doi: 10.1016/j.celrep.2019.09.085

Figure Lengend Snippet: (A and B) ON SACs biolistically labeled with cytosolic tdTomato (magenta) and PSD95-YFP (green), a marker of excitatory synapses, in flat-mounted P20 wild-type (A) and Amigo2 KO (B) retinas. Cells were digitally isolated in Amira for visual clarity. (C and D) Higher magnification view of the insets in (A) (shown in C) and (B) (shown in D). (E and F) Dendrite tracings (magenta) and output of synapse identification (green) for the ON SACs in (A) (shown in E) and in (B) (shown in F). (G) Cumulative distributions of ON SAC dendrite territories in wild-type (37,259 ± 1,361 μm 2 , n = 34 cells, n = 12 retinas) and Amigo2 KO (47,584 ± 1,712 μm 2 , n = 53 cells, n = 16 retinas) mice; p = 5.5 3 10 −5 by Mann-Whitney U test. (H) Cumulative distributions of ON SAC dendrite lengths in wild-type (3,028 ± 86 μm, n = 25 cells, n = 12 retinas) and Amigo2 KO (3,422 ± 85 μm, n = 36 cells, n = 16 retinas) retinas; p = 0.0021 by Mann-Whitney U test. (I and J) Summary data of Sholl analyses for ON SAC branching patterns in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 18 cells, n = 10 retinas) retinas. Distributions of branches as a function of absolute distance from the soma; p = 0.0056 by bootstrapping (I). (J) Distributions of branches as a function of normalized radial distance; p = 0.21 by bootstrapping (J). (K and L) Summary data of the radial distribution of excitatory synapses in ON SAC dendrites in wild-type (n = 16 cells, n = 10 retinas) and Amigo2 KO (n = 16 cells, n = 9 retinas) retinas. Distributions of synapses as a function of absolute distance from the soma; p = 0.012 by bootstrapping (K). Distributions of synapses as a function of normalized radial distance; p = 0.79 by bootstrapping (L). Throughout the figure, ***p < 0.001, **p < 0.01, and ns indicates no significant differences for statistical comparisons.

Article Snippet: We coated gold particles (diameter: 1.6 μm, Bio-Rad) with plasmids encoding cytosolic tdTomato and postsynaptic density protein 95 (PSD95) fused at its C terminus to YFP ( Kerschensteiner et al., 2009 ), or AMIGO2-DDK (Origene).

Techniques: Labeling, Marker, Isolation, MANN-WHITNEY

KEY RESOURCES TABLE

Journal: Cell reports

Article Title: AMIGO2 Scales Dendrite Arbors in the Retina

doi: 10.1016/j.celrep.2019.09.085

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: We coated gold particles (diameter: 1.6 μm, Bio-Rad) with plasmids encoding cytosolic tdTomato and postsynaptic density protein 95 (PSD95) fused at its C terminus to YFP ( Kerschensteiner et al., 2009 ), or AMIGO2-DDK (Origene).

Techniques: Recombinant, Labeling, Software