Claims 26
From the paper Selective control of synaptically-connected circuit elements by all-optical synapses · claims file · checked 2026-10-23: ready
- fig2d-chr2-ctz-excites
In mixed cultures with ChR2(C128S)-expressing postsynaptic neurons, blue light and CTZ both increase spiking, while vehicle has no consistent effect.
We observed a significant difference in spontaneous activity in cultures before and after stimulation by blue light LED and CTZ but no consistent differences in neural activity before and after vehicle (ChR2(C128S), blue light, n = 62, p < 0.0001, CTZ, n = 62, p < 0.0001, vehicle, n = 24, p = 0.6498
Figure 2d; Results, Interluminescence modulates spontaneous neural activity in culture, paragraph 2How it will be judged
As the Methods and the Fig. 2d legend give it, per electrode the spike counts in 5 s before and 5 s after each treatment, a two-tailed Wilcoxon matched-pairs signed-rank test. Reproduced when blue light and CTZ each give p < 0.05 with the median count higher after, and vehicle gives p >= 0.05, with the paper's electrode counts (62, 62, 24). The vehicle part is a claim of absence judged on the same test.
- fig2d-gtacr2-ctz-inhibits
In mixed cultures with hGtACR2-expressing postsynaptic neurons, blue light and CTZ both decrease spiking, while vehicle has no consistent effect.
hGtACR2, blue light, n = 49, p < 0.0001, CTZ, n = 49, p < 0.0001, vehicle, n = 49, p = 0.5594; Wilcoxon matched-pairs signed rank test
Figure 2d; Results, Interluminescence modulates spontaneous neural activity in culture, paragraph 2How it will be judged
As for fig2d-chr2-ctz-excites, with the median count lower after blue light and after CTZ (p < 0.05 each), and vehicle at p >= 0.05, with 49 electrodes in each condition.
- fig3c-uncut-responses
In connected (un-cut) cortical to hippocampal or striatal co-cultures, blue light, presynaptic electrical stimulation and CTZ each increase spiking of the ChR2(C128S)-expressing postsynaptic neurons.
blue light, un-cut n = 63, p < 0.0001, cut n = 58, p < 0.0001; electrical stimulation, un-cut n = 54, p < 0.0001
Figure 3c; Results, Interluminescence requires connections between pre- and postsynaptic neurons, paragraph 3How it will be judged
Per electrode, spikes in 5 s before against 5 s after each treatment, two-tailed Wilcoxon matched-pairs signed-rank test (Methods; Fig. 3c legend). Reproduced when each of the three un-cut conditions gives p < 0.05 with the median higher after.
- fig3c-cut-abolishes-ctz
After the connections between the two populations are cut, blue light still increases postsynaptic spiking, but presynaptic electrical stimulation and CTZ no longer change it.
In contrast, direct electrical stimulation of presynaptic neurons and CTZ-induced bioluminescence failed to alter the excitability of postsynaptic neurons in cut co-cultures
Figure 3c; Results, Interluminescence requires connections between pre- and postsynaptic neurons, paragraph 3How it will be judged
The same test as fig3c-uncut-responses on the cut co-cultures. The absence parts are judged on that test at p < 0.05. Reproduced when blue light gives p < 0.05 with the median higher after, and electrical stimulation and CTZ each give p >= 0.05; the median change is reported either way.
- figs4c-blockers-silence
The synaptic blocker cocktail silences spontaneous activity of the culture, while blue light still activates the opsin-expressing neurons.
silenced spontaneous activity of the entire neural culture on MEAs for several minutes (Supplementary Fig. 4a, b (i), c; n = 27, P < 0.0001; Mann–Whitney test
Supplementary Figure 4c; Results, Interluminescence depends on presynaptic activity and occurs independent of classic synaptic neurotransmission, paragraph 1How it will be judged
The paper's test, a two-tailed Mann-Whitney test on 5 s spike counts (Supplementary Fig. 4c legend; Methods), spontaneous against after SB, and after SB against blue light. Reproduced when both give p < 0.05, with counts lower after SB and higher with blue light. A Wilcoxon signed-rank test on the same electrodes is shown as a variant, as the counts are paired.
- fig4c-sb-ctz-activates
CTZ given with or immediately after the synaptic blockers increases spiking of the opsin-expressing neurons, while vehicle has no effect.
When CTZ was delivered with, or immediately after, addition of synaptic blockers (SB), postsynaptic activation through bioluminescence was robust, while vehicle addition had no effect
Figure 4c; Results, Interluminescence depends on presynaptic activity and occurs independent of classic synaptic neurotransmission, paragraph 1How it will be judged
Two-tailed Mann-Whitney tests on the 5 s spike counts after treatment between groups, as in the Fig. 4c legend. Reproduced when SB + CTZ is higher than both SB alone and SB + vehicle at p < 0.05, and SB alone against SB + vehicle gives p >= 0.05 (the paper's test for absence).
- fig4e-late-ctz-no-effect
CTZ added about 20 s or more after the synaptic blockers has no effect on spiking of the opsin-expressing neurons.
However, addition of CTZ more than 20 s after addition of the synaptic blockers had no effect on spiking of opsin-expressing neurons
Figure 4e(i); Results, Interluminescence depends on presynaptic activity and occurs independent of classic synaptic neurotransmission, paragraph 1How it will be judged
A claim of absence, judged on the paper's test, a two-tailed Mann-Whitney test of the 5 s counts after SB against after CTZ added later (Fig. 4e legend). Reproduced when p >= 0.05; the median difference is reported either way.
- fig4e-stim-then-ctz-activates
During the silent phase after synaptic block, CTZ added immediately after presynaptic electrical stimulation increases spiking of the opsin-expressing neurons, and more than CTZ added later, electrical stimulation alone, or stimulation followed by vehicle.
This interpretation is further supported by the observation that CTZ can induce spiking during the silent phase if it is added immediately following presynaptic electrical stimulation
Figure 4e(iii) and the yellow bars of Figure 4e; Results, Interluminescence depends on presynaptic activity and occurs independent of classic synaptic neurotransmission, paragraph 2How it will be judged
Two-tailed Mann-Whitney tests on the 5 s counts after treatment, as in the Fig. 4e legend. Reproduced when stimulation followed by CTZ is higher than stimulation alone (iii), and higher than each of CTZ added later, stimulation alone and stimulation followed by vehicle (yellow bars), all at p < 0.05. The legend gives one p for the yellow-bar comparison of four groups and does not say how it was made; each pairwise test against the immediate-CTZ group is run, and the claim holds when all three are significant.
- fig4e-stim-alone-no-effect
After synaptic block, presynaptic electrical stimulation alone, or followed by vehicle, does not change spiking of the opsin-expressing neurons.
blockers (after) with electrical stimulation alone (after) or electrical stimulation (after) with immediate vehicle addition (after) had no effect
Figure 4e(ii) and 4e(iv); Results, Interluminescence depends on presynaptic activity and occurs independent of classic synaptic neurotransmission, paragraph 2How it will be judged
A claim of absence, judged on the paper's two-tailed Mann-Whitney tests (Fig. 4e legend). Reproduced when both give p >= 0.05; the median differences are reported. The legend prints p > 0.9999 and the Results p = 0.9999 for (ii); either is read as no effect.
- fig5c-bont-blocks
After 48 h of BoNT treatment, CTZ added immediately after presynaptic electrical stimulation no longer increases postsynaptic spiking, while blue light still does.
CTZ added immediately following electrical stimulation of presynaptic neurons failed to trigger Interluminescence as indicated by the absence of postsynaptic activity in BoNT-treated cultures
Figure 5c; Results, Interluminescence depends on presynaptic vesicle release, paragraph 1How it will be judged
Two-tailed Wilcoxon matched-pairs signed-rank tests on 5 s counts, as in the Fig. 5c legend. The absence part is judged on that test. Reproduced when stimulation against stimulation plus CTZ gives p >= 0.05 and blue light gives p < 0.05 with the median higher during light.
- figs5-bont-reduces-release
BoNT treatment reduces the release of vesicular luciferase into the medium, seen as less bioluminescence in conditioned medium.
In BoNT-treated cultures, the level of luciferase in the media was reduced compared to untreated cultures, consistent with a decrease in vesicle release
Supplementary Figure 5; Results, Interluminescence depends on presynaptic vesicle release, paragraph 1How it will be judged
Shown by one pair of IVIS images and no number or test. Reproduced when the bioluminescence radiance of conditioned medium from BoNT-treated cultures is lower than from untreated ones. The images are not deposited.
- figs6-luciferase-in-dcvs
Luciferase targeted by the POMC signal partly colocalises with the dense-core vesicle marker dopamine beta-hydroxylase, and a considerable fraction lies in other vesicles.
We observed partial colocalization of eGFP and anti-dopamine β-hydroxylase, confirming the presence of sbGLuc in dopamine β-hydroxylase expressing synaptic vesicles, but we also observed sbGLuc-eGFP signals consistent with a considerable fraction of luciferases in other vesicles
Supplementary Figure 6; Results, Interluminescence depends on presynaptic vesicle release, paragraph 2How it will be judged
Shown by representative confocal images and no measure. Reproduced when some eGFP puncta are DBH positive and some are not. The images are not deposited.
- fig6c-green-light-abolishes
In the presence of synaptic blockers, CTZ increases spiking of ChR2(C128S)-expressing neurons, and 590 nm light, which closes the step-function opsin, abolishes that spiking.
590 nm light completely abolished spiking initiated by CTZ
Figure 6c; Results, Interluminescence-mediated activation of postsynaptic neurons requires opsins, paragraph 1How it will be judged
Two-tailed Wilcoxon matched-pairs signed-rank tests on 5 s counts (Fig. 6c legend). The legend does not say which counts each test pairs. Readings, (a) SB + CTZ is before against after adding SB + CTZ, and green light is before against after the 590 nm light; (b) green light is before SB + CTZ against after the 590 nm light. Reproduced when SB + CTZ gives p < 0.05 with counts higher after, and green light gives p < 0.05 with counts lower after, under the reading the source data's columns support. The Results print p = 0.0001 and the legend p < 0.0001 for green light; either matches.
- fig6f-nonfunctional-opsin-no-response
With the non-functional opsin ChR2(C128S)-E97R-D253A in postsynaptic neurons, CTZ does not increase spiking in the presence of synaptic blockers.
In cultures expressing inactive ChR2(C128S)-E97R-D253A in postsynaptic neurons, CTZ generated bioluminescence, but no increase in spiking
Figure 6f; Results, Interluminescence-mediated activation of postsynaptic neurons requires opsins, paragraph 1How it will be judged
A claim of absence, judged on the paper's two-tailed Mann-Whitney test of 5 s counts after SB alone against after SB + CTZ (Fig. 6f legend). Reproduced when p >= 0.05; the median difference is reported.
- figs3-ctz-acts-postsynaptically
In co-cultures, blue light and CTZ increase spiking of the postsynaptic population without changing activity of the presynaptic cortical neurons, and only electrodes that respond to blue light respond to CTZ.
Blue light LED stimulation (which directly activates the postsynaptic Opsin) or CTZ addition (which depends on ongoing spontaneous presynaptic activity) by contrast, increased spiking in ChR2(C128S) expressing postsynaptic neurons but without a change in the activity of cortical presynaptic neurons
Supplementary Figure 3d, 3e; Results, Interluminescence requires connections between pre- and postsynaptic neurons, paragraph 2How it will be judged
Shown by representative recordings with no measure or test. Reproduced when, in co-culture recordings, CTZ raises 5 s spike counts on postsynaptic electrodes and not on presynaptic ones, and no electrode without a blue-light response responds to CTZ. The presynaptic electrodes' counts are not in the Supplementary Data as described, and the recordings are not deposited.
- figs2-cut-disrupts-synchrony
Co-cultures with intact connections fire synchronously across the two populations with a short delay; cutting the connections disrupts the inter-population synchrony.
Representative example of MEA recording of a mature (DIV21) co-culture (i) showing synchronicity in spontaneous intra-population firing activity, but a strongly disrupted inter-population synchronicity
Supplementary Figure 2; Methods, Un-cut vs Cut experimentHow it will be judged
Shown by representative recordings with no measure or test. The recordings are not deposited.
- fig7b-bioluminescence-both-groups
CTZ infusion causes a rise in bioluminescence over barrel cortex in both Opsin (+) and Opsin (-) mice.
In both Opsin (+) mice and Opsin (−) control mice CTZ presentation caused a rise in bioluminescent light production
Figure 7b; Results, Interluminescence in vivo, paragraph 3How it will be judged
The bioluminescence time series of the in vivo pipeline (see the opening comment), for the five imaged mice (2 Opsin (+), 3 Opsin (-), as in the Fig. 7b legend), under ROI readings (o2a) and (o2b). The paper reports no test. Reproduced when, in every imaged mouse, the mean over 0 to 1200 s after onset exceeds the mean over the 180 s before onset by more than the upper bound of the bootstrapped 95% confidence interval of the baseline mean (1 s frames as samples, 10000 resamples as a standard choice, two-sided bound). The Methods give this rule for an electrode contact exceeding baseline, not for the bioluminescence signal; its use here is a choice of this file, and the mean after and before onset are shown for each mouse.
- fig7c-gamma-emerges-opsin-plus
In Opsin (+) mice broad-band gamma oscillations emerge with CTZ, and similar changes are not seen in Opsin (-) mice despite bioluminescence.
In Opsin (+) mice, we observed robust, broad-band gamma oscillation emergence that initiated with CTZ presentation ... In contrast, similar changes were not observed in the Opsin (−) control group, despite robust bioluminescent photon output
Figure 7c; Results, Interluminescence in vivo, paragraph 3How it will be judged
Fig. 7c is a group spectrogram averaged over all contacts. From the in vivo pipeline, the group mean over contacts and mice of P (dB) over 30 to 100 Hz (the Methods' gamma band) and 0 to 1200 s after onset. Reproduced when the Opsin (+) value is above 0 dB and above the Opsin (-) value. The paper gives no threshold for "not observed"; the per-mouse values are reported. Readings (o1) and (o2) are run; the outcome is the one all readings give, and when they differ the claim is blocked as methods_underspecified, with every result shown.
- fig7d-contacts-exceeding-baseline
The gamma increase is seen on about half of the contacts in Opsin (+) mice (15.7 +/- 10.1 SD contacts per mouse) against 2.7 +/- 2.5 in Opsin (-) mice.
were evident on ~half of all electrode contacts in the Opsin (+) group (Fig. 7d : Opsin (+) = 15.7 ± 10.1 SD electrodes/mouse; Opsin (−): M = 2.7 ± 2.5 SD
Figure 7d; Results, Interluminescence in vivo, paragraph 3How it will be judged
Tolerances are the SDs the paper states. A contact exceeds baseline when its mean gamma power (dB) over 0 to 1200 s after onset is above the bootstrapped 95% confidence interval of the mean of its baseline (Methods, Statistical analysis), with the baseline as the 1 s spectrogram steps of the 180 s before onset and 10000 resamples as a standard choice. Open, with the readings run. The band, (a) 30 to 100 Hz (Methods) or (b) 80 to 100 Hz (Fig. 7d legend). The bound, (a) the upper end of a two-sided 95% interval or (b) a one-sided 95th percentile. Plus (o1) and (o2). Reproduced when both means are within tolerance and the Opsin (+) mean is higher. The outcome is the one all readings give; when they differ the claim is blocked as methods_underspecified, with every result shown.
- fig7d-superficial-layers
The gamma increase in Opsin (+) mice is localised to the more superficial and granular layers of the cortical depth.
These Interluminescence gamma increases were localized to more superficial and granular layers across the neocortical depth
Figure 7d; Results, Interluminescence in vivo, paragraph 3How it will be judged
Fig. 7d shows depth profiles of gamma power (dB) across contacts; the paper gives no layer boundaries and no test. Per contact, the Opsin (+) group mean over mice of gamma power (dB) over 0 to 1200 s after onset, ordered by contact depth from the probe map. Open, with the readings run. The band, (a) 80 to 100 Hz (Fig. 7d legend) or (b) 30 to 100 Hz (Methods). The split, (a) the upper half of the kept contacts against the lower half, or (b) the upper two thirds against the deepest third (the probe spans about 790 um from the surface, so its deepest part lies below the granular layer). Reproduced when the upper part has the higher mean under every reading.
- fig7e-gamma-power-electrodes
Across electrodes, gamma power relative to baseline is higher in Opsin (+) than in Opsin (-) mice (mean 0.53 +/- 0.86 SD against -0.10 +/- 0.24 dB; medians 0.34 and -0.17).
Analysis of each metric at the mouse or electrode level showed significantly higher values for the Opsin (+) group (Fig. 7e Mean gamma (dB) power across electrodes: Opsin (+) Mean = 0.53 ± 0.86 SD, Median = 0.34; Opsin (−) Mean = −0.10 ± 0.24 SD, Median = −0.17; KS test p < 0.0001; 1-tailed Mann–Whitney U test U = 4163, n 1 = n 2 = 75, p < 0.0001
Figure 7e; Results, Interluminescence in vivo, paragraph 3How it will be judged
Gamma power from the in vivo pipeline, 30 to 100 Hz (Methods). The KS test is on all contact by time-bin by animal values of each group pooled (1 s spectrogram steps over 0 to 1200 s); the Mann-Whitney test, one-tailed (Opsin (+) greater), is on the 75 time-averaged contact values of each group (Methods, Statistical analysis). Means tolerate the stated SDs, medians 10%. Reproduced when both tests give p < 0.05 in the stated direction and the four values are within tolerance under every combination of readings (o1) and (o2) (when the readings give different outcomes, the claim is blocked as methods_underspecified, with every result shown); partially reproduced when the tests hold and the values do not. Fig. 7d's band (80 to 100 Hz) is shown as a variant.
- fig7e-gamma-events-electrodes
Across electrodes, gamma events after CTZ are more numerous in Opsin (+) than in Opsin (-) mice (mean 213.60 +/- 139.53 SD against 139.28 +/- 72.97; medians 222 and 149).
Mean gamma event number across electrodes: Opsin (+) Mean = 213.60 ± 139.53 SD, Median = 222; Opsin (−) Mean = 139.28 ± 72.97 SD, Median = 149; KS test p < 0.0001; 1-tailed Mann–Whitney U test U = 3609.50, n 1 = n 2 = 75, p < 0.0001
Figure 7e; Results, Interluminescence in vivo, paragraph 3How it will be judged
Gamma events from the in vivo pipeline, counted per contact over the 1200 s window, under readings (o1), (o2), (o3) and (o4). The time bins of the pooled KS test for events are not given. Readings, (a) event counts per 1 s bin, (b) the per-contact totals. The Mann-Whitney test is one-tailed on the 75 per-contact counts of each group. Means tolerate the stated SDs, medians 10%. Reproduced when both tests give p < 0.05 with Opsin (+) higher and the four values are within tolerance under every combination of readings (when the readings give different outcomes, the claim is blocked as methods_underspecified, with every result shown); partially reproduced when the tests hold and the values do not.
- fig7e-gamma-power-mice
At the mouse level, gamma power relative to baseline is higher in Opsin (+) than in Opsin (-) mice (mean 1.16 +/- 1.09 SD against 0.03 +/- 0.18 dB), with every Opsin (+) mouse above every Opsin (-) mouse (U = 9).
Mean gamma power across mice Opsin (+) Mean = 1.16 ± 1.09 SD, Median = 0.88; Opsin (−) Mean = 0.03 ± 0.18 SD, Median = 0.09; 1-tailed Mann–Whitney U test U = 9, n 1 = n 2 = 3, p < 0.05
Figure 7e; Results, Interluminescence in vivo, paragraph 3How it will be judged
Each mouse's value is the 85th percentile of its 25 time-averaged contact values (Methods, Statistical analysis), with the percentile under readings (o6) and alignment under (o1) and (o2). Means tolerate the stated SDs, medians 10%. With 3 against 3 mice, U = 9 means complete separation, and its exact one-tailed p is 1/20 = 0.05, so the test is judged by U. Reproduced when U = 9 with Opsin (+) higher and the four values are within tolerance under every combination of readings (when the readings give different outcomes, the claim is blocked as methods_underspecified, with every result shown); partially reproduced when U = 9 and the values are not.
- fig7e-gamma-events-mice
At the mouse level, gamma events are more numerous in Opsin (+) than in Opsin (-) mice (mean 327.25 +/- 45.58 SD against 199.33 +/- 36.08), with complete separation (U = 9).
Mean gamma event number across mice Opsin (+) Mean = 327.25 ± 45.58 SD, Median = 304.25; Opsin (−) Mean = 199.33 ± 36.08 SD, Median = 203.75; 1-tailed Mann–Whitney U test U = 9, p < 0.05
Figure 7e; Results, Interluminescence in vivo, paragraph 3How it will be judged
As fig7e-gamma-power-mice, on the per-contact gamma event counts, under readings (o1) to (o4) and (o6).
- fig7f-mua-electrodes
Across electrodes, MUA rises more from baseline in Opsin (+) than in Opsin (-) mice (mean 37.14 +/- 45.50 SD against 13.71 +/- 21.44 percent; medians 15.49 and 8.96).
Opsin (+) Mean = 37.14 ± 45.50 SD, Median = 15.49; Opsin (−) Mean = 13.71 ± 21.44 SD, Median = 8.96; KS test p < 0.0001; 2-tailed Mann–Whitney U test U = 3899, n 1 = n 2 = 75, p < 0.0001
Figure 7f; Results, Interluminescence in vivo, paragraph 4How it will be judged
MUA from the in vivo pipeline under readings (o1), (o2) and (o5); the KS test on all 1 s bins over 0 to 1200 s of each group pooled, the two-tailed Mann-Whitney test on the 75 time-averaged contact values of each group (Methods). Means tolerate the stated SDs, medians 10%. Reproduced when both tests give p < 0.05 with Opsin (+) higher and the four values are within tolerance under every combination of readings (when the readings give different outcomes, the claim is blocked as methods_underspecified, with every result shown); partially reproduced when the tests hold and the values do not.
- fig7f-mua-mice-not-significant
At the mouse level the MUA difference is not significant (mean 77.72 +/- 47.40 SD against 27.15 +/- 18.38 percent; U = 9, p > 0.05, two-tailed), though Opsin (+) mice show MUA increases higher than, or equal to, the highest Opsin (-) values.
MUA differences were significant when analyzed at the electrode, but not the mouse, level ... Mean MUA (percent change from baseline) across mice: Opsin (+) Mean = 77.72 ± 47.40 SD, Median = 52.15; Opsin (−) Mean = 27.15 ± 18.38 SD, Median = 27.24; 2-tailed Mann–Whitney U test U = 9, p > 0.05
Figure 7f; Results, Interluminescence in vivo, paragraph 4How it will be judged
A claim of absence, judged on the paper's test, a two-tailed Mann-Whitney U test on each mouse's 85th percentile of time-averaged contact MUA (Methods), under readings (o1), (o2), (o5) and (o6). With 3 against 3 mice the smallest two-tailed exact p is 0.1, so this test cannot reach p < 0.05; the absence part holds whatever the data, and the claim is judged on U and the values. Reproduced when U = 9 with Opsin (+) higher and the four values are within tolerance under every combination of readings (when the readings give different outcomes, the claim is blocked as methods_underspecified, with every result shown); partially reproduced when U is 7 or 8, or U = 9 and the values are not within tolerance.
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