Journal: Nature

Article Title: Human assembloid model of the ascending neural sensory pathway

doi: 10.1038/s41586-025-08808-3

Figure Lengend Snippet: a , Generation of hSeO–hdSpO assembloids. b , Gradual increase in sensory–spinal cord projections in hSeO–hdSpO. c , Quantification of EYFP in the hdSpO side of assembloids ( n = 10 assembloids; three differentiations; Kruskal–Wallis test with Dunn’s test; * P = 0.0267). d , Retrograde rabies viral tracing in hSeO–hdSpO. e , EGFP + tdTomato + cells in hSeO side. f , BRN3A + tdTomato + co-localization in hSeO side. g , Percentage of BRN3A + cells of tdTomato + or Hoechst + cells in hSeO side of assembloids ( n = 6 assembloids; three differentiations; Mann–Whitney test; ** P = 0.0022). h , Patch-clamp recordings with optogenetic stimulation of hSeO neurons. i , oEPSC traces from three trials in the same neuron. j , Generation of hdSpO–hDiO assembloids. k , Gradual increase in spinothalamic projections in hdSpO–hDiO. l , Quantification of EYFP in the hDiO side of assembloids ( n = 10–11 assembloids; three differentiations; Kruskal–Wallis test with Dunn’s test; ** P = 0.0050; *** P = 0.0003). m , Retrograde rabies viral tracing in hdSpO–hDiO. n , EGFP + tdTomato + cells in the hdSpO side of assembloids. o , NK1R + tdTomato + co-localization in the hdSpO side of assembloids. p , Percentage of NK1R + cells of tdTomato + or Hoechst + cells in the hdSpO side of assembloids ( n = 6 assembloids; three differentiations; Mann–Whitney test; ** P = 0.0022). q , Calcium imaging of spinothalamic neurons with retrograde viral tracing in hSeO–hdSpO–hDiO. r , GCaMP6f + dsRedExpress + co-expression in the hdSpO side of assembloids. s , αβ-MeATP-evoked responses and blockade by NBQX and APV (50 μM each). t , Responses to αβ-MeATP with or without NBQX and APV ( n = 74 cells; four assembloids; paired t -test; **** P < 0.0001) on daf 70–88. Data: mean ± s.e.m. Small dots, cells; large dots, assembloids. Scale bars, 100 μm ( b , e (left), k , n (left)), 50 μm ( n (right), r (top)), 30 μm ( e (right)), 20 μm ( f , o ), 10 μm ( r (bottom). Credits: a , d , h , j , m , q adapted from ref. , Springer Nature America, Inc.

Article Snippet: For optogenetic stimulation, whole-field light-emitting diode illumination (460 nm; 5–20 ms duration; maximal power; CoolLED) was applied through a ×40 objective, and oEPSCs were recorded at a holding potential of −70 mV.

Techniques: MANN-WHITNEY, Patch Clamp, Imaging, Expressing

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a . Diagram of closed loop ALM/M1TJ optogenetic perturbation of lick n+2. Photoinhibition (40Hz sinusoid with ramp-down) occurs randomly in 30% of trials following a 120 ms delay after touch onset of lick n+1 and lasts for 150 ms (Methods). b . Scatter plots and kernel density estimations of lick n+1 contact angle with or without ALM/M1TJ photoinhibition of lick n+2 (right, left) and subsequent change in lick shooting angle from lick n+1 to n+2 (n= 4852 control lick pairs, 612 opto lick pairs, 7 sessions, 3 mice). Plotting conventions same as in . c . Similar plot as b , but for lick n+2 contact angle and subsequent change in lick shooting angle from lick n+2 to n+3

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques: Control

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a . Diagram of closed loop optogenetic perturbation of lick n+1. Photoinhibition (40Hz sinusoid with ramp-down) occurs randomly in 30% of trials following a 120 ms delay after touch onset of lick n and lasts for 2 s. b . Schematic of cortical regions targeted for photoinhibition; ALM, anterolateral motor cortex; M1TJ, tongue-jaw primary motor cortex; S1TJ, tongue-jaw primary somatosensory cortex; S1BF, barrel cortex. c . Changes in the rate of licks and touch when inhibiting each of the three cortical areas (columns). Bar plots show mean ± 99% hierarchical bootstrap confidence interval with blue color for licks during optogenetic perturbation and black color for licks without optogenetic perturbation. Rate of lick and touch for each mouse shown in grey (ALM/M1TJ, n = 5 sessions, 3 mice; S1TJ: n = 16 sessions, 4 mice; S1BF, n = 3 sessions, 3 mice). *** p<0.001, two-sided paired shuffle test. c . Probability distributions of shooting angle θ shoot for licks with (blue) or without (black) optogenetic perturbation for each of the three cortical areas (columns) (mean ± s.d). d . Probability distributions of maximum length L max for licks with (blue) or without (black) optogenetic perturbation for each of the three cortical areas (columns) (mean ± s.d).

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques:

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a . Diagram of closed loop optogenetic perturbation of lick n+1. Photoinhibition (40Hz sinusoid with ramp-down) occurs randomly in 30% of trials following a 120 ms delay after touch onset of lick n and lasts for 150 ms (Methods). b . Scatter plot and kernel density estimation of lick n+1 contact angle with or without ALM/M1TJ photoinhibition (right, left) and subsequent change in lick shooting angle (n= 5433 control lick pairs, 464 opto lick pairs, 10 sessions, 3 mice). Colors as in Fig. 1d. ALM, anterolateral motor cortex; M1TJ, tongue-jaw primary motor cortex; S1TJ, tongue-jaw primary somatosensory cortex. *** p<0.001, one-sided paired shuffle test. c . Scatter plot and kernel density estimation of lick n+1 contact angle with or without S1TJ photoinhibition (right, left) and subsequent change in lick shooting angle (n= 8748 control lick pairs, 868 opto lick pairs, 17 sessions, 4 mice). Colors as in .

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques: Control

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a . Diagram of variability in timing of optogenetic perturbation relative to lick n+1.Given the variability in lick cycle duration (~6-7 Hz), the time delay Δt between the onset of optogenetic stimuli (triggered with fixed delay of 120 ms after lick n) and time of lick n+1, n+2, and n+3 can vary. b . Probability distribution of time delay Δt between the onset of optogenetic stimuli and time of lick (n = 72 sessions, 7 mice). Lick n+1 as blue, n+2 as purple, n+3 as orange. Dotted lines indicate onset and offset of optogenetic stimuli. c . Schematic of varying time delay Δt relative to lick n+1 (top) or lick n+2 (bottom). For lick n+1 (top), increasing Δt equals greater overlap between optogenetic stimuli and the early phase of the lick cycle. For lick n+2, decreasing Δt equals greater overlap between optogenetic stimuli and the early phase of the lick cycle. d . Schematic of motor cortex targeted for photoinhibition; ALM, anterolateral motor cortex; M1TJ, tongue-jaw primary motor cortex. e . Changes in the probability of licks and touch when photoinhibiting ALM/M1TJ. Bar plots show mean ± 99% hierarchical bootstrap confidence intervals with blue color for lick n+1 during optogenetic perturbation, purple color for lick n+2 after optogenetic perturbation, and black color for lick n+1 without optogenetic perturbation (ALM/M1TJ, n = 10 sessions, 3 mice). * p<0.05, *** p<0.001, two-sided paired shuffle test. f . Schematic of somatosensory cortex targeted for photoinhibition; S1TJ, tongue-jaw primary somatosensory cortex. g . Same as d , but photoinhibiting S1TJ (S1TJ, n = 17 sessions, 4 mice). h . Probability distributions of shooting angle θ shoot for licks with (blue) or without (black) optogenetic perturbation in ALM/M1TJ (mean ± s.d). i . Magnitude of difference between lick angle and port angle |θ shoot | with respect to variability in ALM/M1TJ photoinhibition timing Δt (mean ± 95% hierarchical bootstrap confidence interval). Colors same as d . j, k . Same as g, h , but with optogenetic perturbation in S1TJ. l . Probability distributions of maximum length L max for licks with (blue) or without (black) optogenetic perturbation in ALM/M1TJ (mean ± s.d). m . Maximum length L max with respect to variability in ALM/M1TJ photoinhibition timing Δt (mean ± 95% hierarchical bootstrap confidence interval). Colors same as d . n, o . Same as k, l , but with optogenetic perturbation in S1TJ.

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques:

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a . Optogenetic perturbation of somatosensory processing in the putative lick aiming algorithm. Neural correlates for lick aiming will be perturbed following disrupted somatosensory processing. b . Time series of the behavioural variables (mean ± 99% bootstrap confidence interval; n = 6619 random walks without optogenetic perturbation in S1TJ, n = 857 random walks with optogenetic perturbations, 21 sessions) for left (red) and right (blue) random walks. Time series are grouped by random walk direction and trial-averaged. Duration of optogenetic perturbation shown in shaded cyan box. c-d . Linear decoding of behavioural variables (columns) from populations recorded in the ALM and M1TJ (rows) with or without optogenetic perturbation in S1TJ (dashed and solid lines, respectively). Displayed traces are results from linear decoding from linear models with appropriate time lags added to spikes; ALM: −50 ms, M1TJ: −50 ms. Traces are grouped by random walk direction and trial-averaged across multiple sessions (ALM, n = 12 sessions; M1TJ: n = 9 sessions). Same plotting conventions as in b , except inter-lick intervals are included. e . (Left) Scatter plot of lick n+1 contact angle and the change from lick n to lick n+2 in port angle decoded from neural activity in ALM (n = 3245 control licks, 554 opto licks, 14 sessions). Licks without S1TJ optogenetic perturbation in grey, licks with S1TJ optogenetic perturbation in blue. Solid line indicates the line of best fit from linear regression. Sign of the lick contact angle flipped for ease of visualization. Pearson’s correlation coefficient r shown in inset. *** p<0.001, one-sided paired shuffle test. (Right) Scatter plot of port angle decoded from neural activity in ALM at lick n+1 and actual lick angle of lick n+2 n +2 . Same plotting conventions as in left panel. f . Similar scatter plot as e , but change in port angle decoded from neural activity and port angle decoded from neural activity in M1TJ (n = 2723 control licks, 412 opto licks, 9 sessions).

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques: Activity Assay, Control

Journal: bioRxiv

Article Title: Rapid integration of somatosensory feedback in planning tongue movements

doi: 10.64898/2026.03.03.709371

Figure Lengend Snippet: a , b . Example single unit responses from ALM and M1TJ recordings without ( a ) or with ( b ) S1TJ photoinhibition. Duration of optogenetic perturbation in S1TJ indicated with cyan bar. Same plotting conventions as . c . Single-trial decoding of behavioral variables (rows; black traces) from 24 simultaneously recorded M1TJ units with S1TJ photoinhibition for right (top, blue) and left (bottom, red) random walks. Same plotting conventions as .

Article Snippet: Optogenetic stimuli were applied using a pair of 470 nm LEDs (M595F2, Thorlabs) via an optic fiber (M98L01, Thorlabs) positioned over target areas with approximately 1 mm radius (ALM/M1TJ, AP 2.5, ML ±1.5 mm; S1TJ, AP 0.5, ML ±3.7 mm, 30 deg angle from vertical; S1BF, AP −1.5, ML ±3.5 mm).

Techniques: