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Journal: bioRxiv
Article Title: Tuning metaplasticity in the adult visual cortex using flickering light
doi: 10.64898/2026.02.16.706197
Figure Lengend Snippet: a-c , Single-unit recordings reveal that the ocular dominance shift caused by LTMD during the critical period is reversed by exposure to 40 Hz light-flicker. a , Experimental timeline. b , Cumulative distribution of ODI values for visually responsive single-units in V1b in mice reared normally (NR) or subjected to LTMD. LTMD animals were either untreated or exposed to either 40 Hz or 60 Hz light-flicker. The shift in ODI caused by LTMD is apparent in untreated and 60 Hz groups, but reversed in the 40 Hz group (KS – test with Bonferroni correction: NR vs LTMD: p < 0.001; NR vs LTMD + 40 Hz: p = 0.71688; NR vs LTMD + 60 HZ: p < 0.001; LTMD vs LTMD + 40 Hz: p < 0.001; LTMD vs LTMD + 60 Hz: p = 0.56635; 40 Hz vs LTMD + 60 Hz: p < 0.01). c , Group comparisons (averaged unit ODI values per animal) demonstrate that the OD shift produced by LTMD is completely reversed in animals receiving 40 Hz light-flicker treatment (NR: 0.2694 ± 0.04277; LTMD: 0.01600 ± 0.02450; LTMD + 40 Hz: 0.2509 ± 0.03403; LTMD + 60 Hz: 0.06587 ± 0.03974; One-way ANOVA with Tukey’s test for multiple comparisons, p < 0.001; NR vs LTMD: p = 0.0052; NR vs LTMD + 40 Hz: p = 0.9860; NR vs LTMD + 60 Hz: p = 0.0060; LTMD vs LTMD + 40 Hz: p = 0.0120; LTMD vs LTMD + 60 Hz: p = 0.8600; LTMD + 40 Hz vs LTMD + 60 Hz: p = 0.0172). d-g , The pole descent cliff task (PDCT) reveals visual deficits after LTMD that are reversed after receiving 40 Hz light-flicker. d , Experimental timeline for PDCT experiments. e , Experimental setup. Mice were positioned on top of a pole with head facing downward and the time taken to descend and dismount was recorded. f , LTMD animals have increased pole descent time when compared to NR mice (NR: 7.625 ± 0.498 seconds, LTMD: 11.8 ± 1.181 seconds, Mann-Whitney test: p = 0.00882). g , LTMD mice exposed to light-flicker at 40 Hz have descent times comparable to NR levels whereas the descent time of LTMD + 60 Hz is comparable to LTMD animals (NR: 7.533 ± 0.6389 sec, LTMD + CL: 13.14 ± 1.119 sec, LTMD + 40 Hz: 8.700 ± 0.9976 sec; LTMD + 60 Hz: 13.00 ± 1.317 sec, one-way ANOVA with Tukey’s test for multiple comparisons, p < 0.001; NR vs LTMD + CL: p = 0.0035, NR vs LTMD + 40 Hz: p = 0.8475, NR vs LTMD + 60 Hz: p = 0.0039, LTMD + CL vs LTMD + 40 Hz: p = 0.0179, LTMD + CL vs LTMD + 60 Hz: p = 0.9997, LTMD + 40 Hz vs LTMD + 60 Hz: p = 0.0199). *p < 0.05; **p < 0.01.
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Journal: bioRxiv
Article Title: Tuning metaplasticity in the adult visual cortex using flickering light
doi: 10.64898/2026.02.16.706197
Figure Lengend Snippet: a , Experimental timeline. b , Representative images and summary graph demonstrating increased counts of Iba1+ microglia in V1b of mice exposed to 60 Hz light-flicker compared to CL or 40 Hz light-flicker (n= 4-5 per group; CL: 3.673 ± 0.420; 40 Hz: 3.958 ± 0.289; 60 Hz: 4.892 ± 0.205; one-way ANOVA with Tukey’s test for multiple comparisons, p = 0.0365; CL vs 40 Hz: p = 0.8019; CL vs 60 Hz: p = 0.00395; 40 Hz vs 60 Hz: p = 0.1176). Scale bar 100µm. c , Representative images and summary graph demonstrating elevated CD11b expression in Iba1+ microglia in V1 of mice exposed to 60 Hz light-flicker as compared to CL or 40 Hz light-flicker stimulation (CL: 1.000 ± 0.056; 40 Hz: 0.900 ± 0.063; 60 Hz: 1.383 ± 0.127; one-way ANOVA with Tukey’s test for multiple comparisons, p = 0.0111; CL vs 40 Hz: p = 0.769; CL vs 60 Hz: p = 0.043; 40 Hz vs 60 Hz: p = 0.013). Scale bar 10µm. d , Representative images and summary graph demonstrating reduced aggrecan levels in V1b of mice exposed to 60 Hz light-flicker compared to CL or 40 Hz light-flicker stimulation (CL: 1.000 ± 0.083; 40 Hz: 0.9730 ± 0.076; 60 Hz: 0.477 ± 0.055; one-way ANOVA with Tukey’s test for multiple comparisons, p = 0.0003; CL vs 40 Hz: p = 0.9621; CL vs 60 Hz: p = 0.0007; 40 Hz vs 60 Hz: p = 0.001). Scale bar 20µm. Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.
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Techniques: Expressing
Journal: bioRxiv
Article Title: Tuning metaplasticity in the adult visual cortex using flickering light
doi: 10.64898/2026.02.16.706197
Figure Lengend Snippet: a , Experimental timeline. In separate experiments, effects of MD were measured using acute single unit recordings with translaminar electrodes or VEPs from chronically implanted electrodes. b , Recordings were performed in V1b contralateral to the previously deprived eye (blue) in awake head-fixed mice viewing grating stimuli. c , Cumulative distribution of ODI values for single-units recorded across all layers of V1b in mice exposed to CL (n = 7), 40 Hz (n = 9), or 60 Hz (n = 8) flicker prior to 4 days of MD. Neurons from animals receiving either 40 Hz or 60 Hz light flicker show a statistically significant leftward shift in their ODI as compared to CL (control) animals. KS test with Bonferroni correction: CL vs 40 Hz: **p = 0.0014; CL vs 60 Hz: ****p < 0.0001; 40 Hz vs 60 Hz: p = 0.7592. d , Group comparisons (averaged unit ODI values per animal) indicate a statistically significant OD shift in animals receiving light flicker (CL: 0.2031 ± 0.053037, 40 Hz: 0.03926 ± 0.03970, 60 Hz: 0.02976 ± 0.05470; one-way ANOVA with Dunnett’s multiple comparisons test: p = 0.0386; CL vs 40 Hz: P = 0.0487, CL vs 60 Hz: p = 0.0420. e , Amplitudes of VEPs elicited through the ipsilateral (yellow) and contralateral (blue) eyes were not significantly altered by treatment with CL, 40 Hz, or 60 Hz flicker. f , ODI values from VEP recordings were unchanged by any treatment but shifted by MD in the flicker groups. Comparing ODI values across timepoints reveals no significant changes for the CL group (n = 17; Pre-Treat: 0.3634 ± 0.03115, Post-Treat: 0.3625 ± 0.03105, Post-MD: 0.2532 ± 0.03980, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test, p = 0.0431; Pre-Treat vs Post-Treat: p = 0.9995, Pre-Treat vs Post-MD: p = 0.0949, Post-Treat vs Post-MD: p = 0.1270) whereas both 40 Hz and 60 Hz groups show a significant reduction in ODI values at the Post-MD timepoint (40 Hz, n = 17; Pre-Treat: 0.3769 ± 0.03648, Post-Treat: 0.3965 ± 0.04076, Post-MD: 0.01738 ± 0.05676, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test, p < 0.0001; Pre-Treat vs Post-Treat: p = 0.7329, Pre-Treat vs Post-MD: p = 0.0002, Post-Treat vs Post-MD: p < 0.0001; 60 Hz, n = 15; Pre-Treat: 0.4247 ± 0.02797, Post-Treat: 0.3026 ± 0.05977, Post-MD: -0.2434 ± 0.05722, repeated-measures one-way ANOVA with Tukey’s multiple comparisons test, p < 0.0001; Pre-Treat vs Post-Treat: p = 0.1175, Pre-Treat vs Post-MD: p < 0.0001, Post-Treat vs Post-MD: p < 0.0001). g , Comparing the change in ODI from pre- to post-MD shows little effect of prior exposure to CL, but a significant effect of light-flicker. (CL: -0.1110 ± 0.04909, 40 Hz: -0.3302 ± 0.07826, 60 Hz: -0.6681 ± 0.05831, one-way ANOVA with Tukey’s multiple comparisons test, p < 0.0001; CL vs 40 Hz: p = 0.0428, CL vs 60 Hz: p < 0.0001, 40 Hz vs 60 Hz: p = 0.0016). h , Grouped average VEP waveforms for contralateral (blue) and ipsilateral (yellow) eyes across treatment condition and MD timepoint. Scale bar: 100 µV, 50 msec. i , Change in contralateral (deprived) eye VEP magnitude after MD (CL, 94.47 ± 8.293 %; 40 Hz, 80.02 ± 8.123 %; 60 Hz, 53.47 ± 9.258 %, one-way ANOVA with Tukey’s multiple comparison test, p = 0.0059; CL vs 40 Hz: p = 0.4466, CL vs 60 Hz: p = 0.0044, 40 Hz vs 60 Hz: p = 0.0864). j , Change in ipsilateral (non-deprived) eye VEP magnitude after MD (CL, 121.4 ± 13.13 %; 40 Hz, 182.2 ± 16.59 %; 60 Hz, 152.6 ± 18.16 %, one-way ANOVA with Tukey’s multiple comparison test, p = 0.0302; CL vs 40 Hz: p = 0.0227, CL vs 60 Hz: p = 0.3659, 40 Hz vs 60 Hz: p = 0.4055). Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001. k , Summary of the different responses to 4d MD in the 3 groups.
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Techniques: Control, Comparison
Journal: bioRxiv
Article Title: Tuning metaplasticity in the adult visual cortex using flickering light
doi: 10.64898/2026.02.16.706197
Figure Lengend Snippet: a , Representative Sholl overlay (concentric radii) on skeletonized microglia and summary graph ( b ) demonstrating increased ramification index in V1 of mice exposed to 60 and 40 Hz light-flicker compared to CL (n= 4-5 per group; CL: 2.661 ± 0.079; 40 Hz: 3.766 ± 0.082; 60 Hz: 4.175 ± 0.290; one-way ANOVA with Tukey’s test for multiple comparisons: p = 0.0009; CL vs 40 Hz: p = 0.008; CL vs 60 Hz: p = 0.00008; 40 Hz vs 60 Hz: p = 0.366). c , Summary graphs demonstrating increased soma size in V1 microglia of mice exposed to 60 and 40 Hz light-flicker compared to CL (CL: 39.84 ± 0.585; 40 Hz: 45.77 ± 2.082; 60 Hz: 46.41 ± 0.757; one-way ANOVA with Tukey’s test for multiple comparisons: p = 0.0079; CL vs 40 Hz: p = 0.023; CL vs 60 Hz: p = 0.009; 40 Hz vs 60 Hz: p = 0.928). d , Summary graphs demonstrating increased Iba-1 intensity in V1 microglia of mice exposed to 60 Hz light-flicker as compared to CL or 40 Hz light-flicker stimulation (CL: 1.000 ± 0.071; 40 Hz: 1.100 ± 0.139; 60 Hz: 1.563 ± 0.130; one-way ANOVA with Tukey’s test for multiple comparisons: p = 0.0149; CL vs 40 Hz: p = 0.840; CL vs 60 Hz: p = 0.0185; 40 Hz vs 60 Hz: p = 0.0494). e , Representative images and summary graphs ( f ) demonstrating unchanged PV+ cell counts across groups (n= 5 per group; CL: 2.180 ± 0.060; 40 Hz: 2.116 ± 0.063; 60 Hz: 2.059 ± 0.099; one-way ANOVA: p = 0.546). Scale bar 100µm. g , A reduced percentage of PV+ cells surrounded by aggrecan (CL: 53.18 ± 1.737; 40 Hz: 50.11 ± 3.104; 60 Hz: 43.45 ± 0.432; one-way ANOVA with Tukey’s test for multiple comparisons: p = 0.0175; CL vs 40 Hz: 0.463; CL vs 60 Hz: 0.016; 40 Hz vs 60 Hz: 0.098) and h , reduced aggrecan-covered area in V1 of mice exposed to 60 Hz light-flicker compared to CL or 40 Hz light-flicker stimulation (CL: 101.2 ± 8.637; 40 Hz: 84.41 ± 4.289; 60 Hz: 77.16 ± 4.650; one-way ANOVA with Tukey’s test for multiple comparisons: p = 0.0474; CL vs 40 Hz: 0.176; CL vs 60 Hz: 0.043; 40 Hz vs 60 Hz: 0.693). Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.
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