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. 2017 Mar 31;8:14908. doi: 10.1038/ncomms14908

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Copyright © 2017, The Author(s)

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(a) Responses of a DRN-projecting RGC to looming stimulation. Left, an expanding dark disk on a grey background used as a looming stimulus; Centre, example of physiological responses of a DRN-projecting RGC to looming stimulus; Right, DRN-projecting RGC filled with neurobiotin after recording physiological responses to looming stimulus. (b) Design of immunotoxin anti-CTB-SAP: IgG antibody generated against CTB conjugated to saporin (SAP). Immunotoxin binds to CTB (retrogradely transported to the soma and delivered to the cell surface via transcytosis) and enters the cell through endocytosis. SAP kills the cell by inactivating ribosomes. (c) Examples of CTB-labelled DRN-projecting RGCs in retinal whole mounts after intraocular PBS (VEH) or anti-CTB-SAP (2 μg per eye) injection. CTB-labelled DRN-projecting RGCs are enhanced in the lower panels. (d) Distribution of CTB-labelled DRN-projecting RGCs (enhanced) in retinal whole mounts 14 days after intraocular vehicle (left) or anti-CTB-SAP (right) injections. S: Superior, I: Inferior, N: Nasal and T: Temporal. (e) Quantification of DRN-projecting RGCs in vehicle (VEH) and anti-CTB-SAP (CTB-SAP)-treated animals. (n=8 retinas per group), one-way ANOVA; ***P<0.0001. (f) Schematic of looming animation and testing arena. (g) Representative traces of animal movement during looming stimulation (10.75 s) in VEH and anti-CTB-SAP groups; time in corner is duration of trace from start to end. (h) Duration from start to end points, peak speed during and after looming stimulation in VEH and anti-CTB-SAP (CTB-SAP)-treated animals. One-way ANOVA; ***P<0.0001; ns=no significant difference. (i) The optomotor response of animals treated with VEH or anti-CTB-SAP (CTB-SAP) tested under photopic and scotopic conditions; the responses in both animal groups under both conditions were similar. Scale bars: (a,c) 50 μm; (d) 1 mm. Data presented as mean±s.e.m.