Figure 4.
Role of Tenm3+ AC GABAergic Inhibition in RGC Orientation Selectivity
(A–E) Immunostaining showing the expression of γ-aminobutyric acid (GABA; A), parvalbumin (B), tyrosine hydroxylase (TH; C), choline acetyltransferase (ChAT; D), and glycine (E) (all in magenta) in 3-dpf Tg(tenm3:Gal4;UAS:GCaMP5) larvae, where tenm3+ ACs are labeled with GCaMP5 (green). Yellow arrowheads indicate neurites of TH+ interplexiform ACs (C). INL, inner nuclear layer; GCL, ganglion cell layer; OPL, outer plexiform layer; IPL, inner plexiform layer. Scale bars, 20 μm.
(F) Percentage of GCaMP5+ cells co-localizing with antigen+ cells (mean ± SD). GABA, n = 13 retinae; Parvalbumin, n = 10 retinae; TH, n = 9 retinae; ChAT, n = 10 retinae; glycine, n = 5 retinae.
(G and H) Average number (G) and relative frequency (H) of DS, OS, visually responsive, and non-DS/non-OS voxels per Z-plane in 4-dpf Tg(isl2b:Gal4;UAS:SyGCaMP3) larvae (n = 15 larvae) before (control) and after (picrotoxin) the application of picrotoxin (100 μM). Error bars, ±SEM. ∗∗p < 0.01, ∗∗∗p < 0.001, paired two-tailed Student’s t test.
(I and J) Cumulative distributions of DSI values (R2 > 0) across voxels with OSI < 0.5 (I) and OSI values (R2 > 0) across voxels with DSI < 0.5 (J) before (control) and after (picrotoxin) the application of picrotoxin (100 μM). ∗∗∗p < 0.001, two-sample Kolmogorov-Smirnov test.
(K and L) Cumulative histograms summarizing the incidence of preferred angles for identified DS (K) and OS voxels (L) in 4-dpf larvae (n = 15 larvae) before (control) and after (picrotoxin) the application of picrotoxin (100 μM). Overlaid curves are the fitted Gaussian distributions for each DS or OS subtype. Polar plots illustrate the mean (+1 SD) normalized response profiles for each DS or OS subtype.