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CNE_22802_sm_suppinfofig1.tif6563KSupporting Figure 1: (Magenta-green version of Figure 3 for the assistance of color-blind readers.) Examples of different types of RGCs found in retinas from rd1/Thy1-GFP transgenics. Each cell is shown in whole mount (top part of the panels) and in vertical view after computer rotation (bottom part of the panels). In vertical views, the red signal is ethidium nuclear staining. A1 and A2 cells (A and B) have large-size dendritic trees, while B1 and B2 cells (C and D) have smaller arbors. A and B RGCs of the mouse can be considered the equivalent of α and β types of the cat. The C1 and C3 cells shown here (E and F) stratify in the innermost tier of the IPL, while the C6 cell (G) reaches the outer part of the IPL. Among C3 RGCs, a subset is positive for melanopsin. C6 and D2 types comprise directional selective ganglion cells. The cell labeled as D2 (H) is bistratified and therefore is a putative ON-OFF GC.
CNE_22802_sm_suppinfofig2.tif3122KSupporting Figure 2: (Magenta-green version of Figure 3 for the assistance of color-blind readers.) Examples of undersized (A, C) and normal (B, D) RGCs belonging to the A2 inner and outer types. A type cells have a round soma with an average of 22 μm, an average dendritic field diameter of 300μm, and a radiate pattern of 4-7 primary branching repeatedly proximal to the soma. Cells shown here are from rd1/Thy1-GFP mice aged 9 months. In this and in the following images, each cell is shown in whole mount view and, on the bottom, after computer vertical rotation, illustrating the level of dendritic stratification in the IPL. Red signal: ethidium nuclear staining.
CNE_22802_sm_suppinfofig3.tif6111KSupporting Figure 3: (Magenta-green version of Figure 5 for the assistance of color-blind readers.) Examples of undersized (A, C, E, G) and normal (B, D, F, H) RGCs belonging to the C1, C2 (inner and outer) and B1 types. C type RGCs have an average soma diameter of 16 μm, and dendritic field diameter of 241 μm. They exhibit smooth, small-caliber, recursive dendrites extending from large primary ones. C1 cells stratify mostly in the inner IPL (62% ± 15%). C2 cells have a morphology similar to that of C1 cells but with more curved and denser dendrites. B1 RGCs have an average soma diameter of 14 μm and radially branching dendrites, ramified in the outer IPL close to the middle. B1 cells have an average dendritic field diameter of 173 μm, thus comprising the GCs with the smallest dendritic tree area of the mouse retina.
CNE_22802_sm_suppinfofig4.tif11606KSupporting Figure 4: (Magenta-green version of Figure 6 for the assistance of color-blind readers.) In this and the following 3 panels, RGCs are displayed as whole mounts in the topmost image, as computer vertical rotations in the middle image and as Neurolucida drawings in the bottom panel. A, B: GFP positive GCs belonging to the B3 outer types. at young (A) and older (B) ages. Red signal: ethidium nuclear staining. C, D: Neurolucida-traced dendrites of different orders are shown in different colors. E: diagrams show the analysis of the three indicators of dendritic tree complexity: dendritic field area, total number of nodes and total dendritic tree length. In this and in the following three figures, black asterisks refer to the statistic comparison between undersized RGCs from rd1 mice and RGCs from and wt animals, while gray asterisks refer to the comparison between undersized RGCs from rd1 mice and normal-size RGCs from mice of the same strain. In this and in the following pictures: Data are from groups of 5-10 cells for each age, for each strain of mice. Student t test. * p 0.01<p<0.05; ** p<0.001<p<0.01; *** p<0.001.
CNE_22802_sm_suppinfofig5.tif11607KSupporting Figure 5: (Magenta-green version of Figure 7 for the assistance of color-blind readers.) A-D: B3 inner cells with smaller dendritic tree areas and increased number of nodes (E).
CNE_22802_sm_suppinfofig6.tif11733KSupporting Figure 6: (Magenta-green version of Figure 8 for the assistance of color-blind readers.) A-D: Examples of C2 outer RGCs from rd1 mice showing dendritic trees of smaller areas (E).
CNE_22802_sm_suppinfofig7.tif11369KSupporting Figure 7: (Magenta-green version of Figure 9 for the assistance of color-blind readers.) A-D: Examples of C2 inner RGCs from rd1 mice with smaller dendritic tree areas (E)
CNE_22802_sm_suppinfofig8.tif10981KSupporting Figure 8: (Magenta-green version of Figure 10 for the assistance of color-blind readers.) A: melanopsin positive GC.from the retina of one rd1/Thy1-GFP mice. M1 type. B: Histograms of cell body diameters of melanopsin-positive GCs in retinas from rd1-Thy1/GFP (solid line) and wt (dashed line) mice. The average diameter is smaller in the rd1 mutant. The average diameter of melanopsin positive wt GCs is in agreement with published data (Berson et al.). Student t test; *** :p<0.005. C: Neurolucida drawing of one M1 type melanopsin-positive GC from the retina of one rd1/Thy1-GFP mouse. Different colors represent different branching orders of the dendrites. D:. Dendritic tree diameter distribution of 14 M1 type melanopsin GCs from the rd1/Thy1-GFP mouse. The study of (Berson et al.) reports a minimum diameter of about 190 μm for all melanopsin GCs, while here 9 out of 14 cells have diameters smaller < 180 μm.
CNE_22802_sm_suppinfofig9.tif4989KSupporting Figure 9: (Magenta-green version of Figure 11 for the assistance of color-blind readers.) A GFP-positive GC (arrow in A) from the retina of one rd1/Thy1-GFP mouse. This cell stratifies in the innermost half of the IPL (as visible in the side view shown in B) and has the typical morphology of a B3 GC. However, the dendritic tree diameter is 30% smaller than normal. The cell is also positive after SMI-32 antibodies immunostaining (arrow in C).
CNE_22802_sm_suppinfofig10.tif9654KSupporting Figure 10: (Magenta-green version of Figure 12 for the assistance of color-blind readers.) A and B: Two examples of GFP-positive GCs from rd1/Thy1-GFP mice, labeled with Nav1.6 antibodies (red staining). The AISs are double labeled. C and D: Diagrams of average AIS length (C) and average AIS distance from cell soma (B) in rd1 and wt young (1 month) and adult (1 yr) mice.

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