Fig. 8

Genetic deficiency in Ifng or Tnf exacerbates retinal neurodegeneration and visual dysfunction in an LPS/NMDA-induced injury model. a Targeting strategy for generating Ifng^-/- and Tnf ^+/- mice. b Experimental timeline: Mice received intraperitoneal LPS (0.2 mg/kg) followed 2 h later by intravitreal NMDA (1.5 µL, 5 mM); analyses were performed 48 h post-NMDA. c Light‒dark box test (LDBT) showing reduced time spent in the light compartment by Ifng^-/- mice compared with WT controls (n = 4 WT, Ifng^-/- per group). d Representative visual evoked potential (VEP) waveforms and quantification of N1‒P1 amplitudes showing attenuation in Ifng^-/- mice (n = 4 WT, Ifng^-/- per group). e Representative scotopic electroretinogram (ERG) waveforms and quantification of a-wave and b-wave amplitudes showing a reduction in Ifng^-/- mice (n = 4 WT, Ifng^-/- per group). f cross immune cell typeBrn3a (RGC marker) and quantification showing reduced RGC density in Ifng⁻/⁻ mice (n = 4 WT, Ifng^-/- per group). Scale bar: 20 µm Scale bar: 50 µm. g–k Representative retinal cryosections and quantification of fluorescence intensity for g βIII-tubulin (neuronal marker) and Brn3a (RGCs in the GCL), h PSD95 and Synaptophysin (synaptic markers in the IPL), i representative retinal whole mounts stained for GFAP (astrocyte marker) and immunofluorescence and fluorescence intensity quantification of GFAP in Ifng^-/- mice (n = 4 WT, Ifng^-/- per group). Scale bar: 20 µm; (j) IBA1 (a microglial marker) and k CaMKII (an activity marker) in WT and Ifng^-/- mice (n = 4 WT, Ifng^-/- per group). Scale bars: 20 µm. l LDBT showing reduced time in light for the Tnf ^+/- mice compared with the WT mice (n = 4 WT, Tnf ^+/- per group). m Representative visual evoked potential (VEP) waveforms and quantification of N1‒P1 amplitudes showing attenuation in Tnf^-/- mice (n = 4 WT, Tnf^-/- per group). n ERG analysis showing reduced amplitudes in Tnf ^+/- mice (n = 4 WT, Tnf ^+/- per group). o RGC quantification from Brn3a-stained whole mounts showing loss in Tnf ^+/- mice (n = 4 WT, Tnf ^+/- per group). Scale bar: 20 µm; scale bar: 50 µm. p–t Representative retinal cryosections and quantification of fluorescence intensity: p βIII-tubulin (neuronal marker) and Brn3a (RGCs in the GCL), q PSD95 and Synaptophysin (synaptic markers in the IPL), r representative retinal whole mounts stained for GFAP (astrocyte marker) and immunofluorescence and fluorescence intensity quantification of GFAP in Ifng^-/- mice (n = 4 WT, Ifng^-/- per group). Scale bar: 20 µm Scale bar: 50 µm; s IBA1 (microglia marker), t CaMKII (activity marker) in WT vs Tnf ^+/- mice (n = 4 WT, Tnf ^+/- per group). Scale bars: 20 µm. The data are presented as the means ± SEMs. Statistical significance was determined by unpaired Student’s t test (P < 0.05, P < 0.01, P < 0.001). GCL ganglion cell layer, IPL inner plexiform layer