grab sensor excitation  (Thorlabs)

Journal: iScience

Article Title: The GPCR Smoothened on cholinergic interneurons modulates dopamine-associated acetylcholine dynamics, learning, and effort management

doi: 10.1016/j.isci.2026.115324

Figure Lengend Snippet: Smo on CIN modulates ACh inhibition following repeated DAN stimulation (A) Left: Viral strategy for simultaneous dopamine neuron (DAN) axon terminal stimulation and G protein-coupled receptor-based sensor for dopamine (GRAB DA) or acetylcholine (GRAB ACh) recording in the dorsolateral striatum. Right: Representative traces across four laser pulse trains with session averages. (B) GRAB sensor and ChrimsonR-tdTomato expression in the dorsolateral striatum and substantia nigra pars compacta. Scale bars = 500μm. (C) Daily stimulation protocol in ChATCre +/− or Smo L/L :ChATCre +/− mice with or without Smoothened agonist (SAG). (D) Dopamine (DA) signals aligned to laser onset on days 1 and 7 in ChATCre +/- mice ( n = 7). A single SAG dose at 20 Mg/Kg was administered on day 8 to assess effects on DA release. (E) Quantification of DA release area under the curve (AUC) from (D) reported as percent of day 1 ( n = 7; repeated-measures one-way ANOVA: F(1.378, 8.269) = 0.03, p > 0.05). (F) ACh signal on days 1 and 7 in ChATCre +/− mice ( n = 7). (G) Same as (F), in Smo L/L :ChATCre +/− mice ( n = 6). (H) Comparison of GRAB ACh AUC on days 1 and 7 in ChATCre +/− mice ( n = 7 per day; paired two-tailed Student’s t test, ∗∗ p < 0.01). (I) Comparison of GRAB ACh AUC on days 1 and 7 in Smo L/L :ChATCre +/− mice ( n = 6 per day; paired two-tailed Student’s t test, p > 0.05). (J–K) ACh signal in SAG-treated mice on days 1 and 7 ( n = 7–8 per genotype). (L) Percent change in ACh dip amplitude in SAG-treated animals relative to untreated control average for each genotype ( n = 6–7; paired two-tailed Student’s t test; ∗∗ p < 0.01). (M) ACh dip AUC across stimulation days with or without SAG treatment in ChATCre +/− ( n = 7–8 per day; two-way repeated-measures ANOVA: day effect, F(2.902, 37.72) = 6.4, ∗∗ p < 0.01; Pharmacology effect, F(1, 13) = 23.47, ∗∗∗ p < 0.001; day × pharmacology interaction, F(4, 52) = 2.03, p > 0.05) and Smo L/L :ChATCre +/− mice ( n = 6–7 per day; two-way repeated-measures ANOVA: day effect, F(2.615, 28.76) = 0.57, p > 0.05; pharmacology effect, F(1, 11) = 1.41, p > 0.05; day × pharmacology interaction, F(4, 44) = 2.18, p > 0.05). (N) ACh dip duration across stimulation days with or without SAG treatment in ChATCre +/− ( n = 7–8 per day; two-way repeated-measures ANOVA: Day effect, F(3.491, 45.39) = 4.81, ∗∗ p < 0.01; Pharmacology effect, F(1, 13) = 5.52, ∗ p < 0.05; day × pharmacology interaction, F(4, 52) = 1.62, p > 0.05) and Smo L/L :ChATCre +/− mice ( n = 6–7 per day; two-way repeated-measures ANOVA: day effect, F(2.785, 30.64) = 0.35, p > 0.05; pharmacology effect, F(1, 11) = 0.57, p > 0.05; day × pharmacology interaction, F(4, 44) = 1.64, p > 0.05).

Article Snippet: ChrimsonR excitation, GRAB sensor excitation, isosbestic excitation (405 nm), and signal collection were all performed through a single patch cord (MFP_200/220/LWMJ-0.37_1m_FCM-MF1.25(F)_LAF; Doric Lenses), connected to the animal’s implanted optical fiber via an interconnect (ADAL3; Thorlabs), and to the sample (S) port of the Mini Cube.

Techniques: Inhibition, Expressing, Comparison, Two Tailed Test, Control

Journal: iScience

Article Title: The GPCR Smoothened on cholinergic interneurons modulates dopamine-associated acetylcholine dynamics, learning, and effort management

doi: 10.1016/j.isci.2026.115324

Figure Lengend Snippet: Smo on CIN modulates ACh inhibition coincident with endogenous DAN bursts (A) Left: Viral strategy for simultaneous G protein-coupled receptor-based acetylcholine and dopamine sensor (GRAB ACh and GRAB DA) recordings in the dorsolateral striatum. Right: Representative traces from a single mouse, with session-averaged GRAB DA event-aligned ACh profiles. (B) GRAB sensor expression in the striatum. Scale bars = 500μm. (C) Recording protocol for control and Smo L/L :ChATCre +/− mice. (D) Frequency of detected DA events with detection threshold set to 2, 3, or 4 MAD ( n = 16 per MAD threshold). (E) Average Spontaneous DA events stratified by amplitude ( top ) and coincident ACh signal ( bottom ) from control and Smo L/L :ChATCre +/− mice ( n = 7–9 per condition). (F) Area under the curve (AUC) for ACh bursts preceding DA events in (E) ( n = 7–9 per condition; two-way repeated measures ANOVA: genotype effect, F(1, 14) = 2.07, p > 0.05; coincident DA effect, F(1.143, 16.01) = 1.02, p > 0.05; genotype × coincident DA interaction, F(2, 28) = 0.84, p > 0.05). (G) AUC for ACh inhibition following DA events in (E) ( n = 7–9 per condition; two-way repeated measures ANOVA: genotype effect, F(1, 14) = 3.22, p > 0.05; coincident DA effect, F(1.555, 21.77) = 5.46, p < 0.05; genotype × coincident DA interaction, F(2, 28) = 5.80, p < 0.01; post hoc Šídák’s multiple comparisons test: ∗∗ p < 0.01). (H) ACh inhibition duration in (E) ( n = 7–9 per condition; two-way repeated measures ANOVA: genotype effect, F(1, 14) = 4.70, ∗ p < 0.05; coincident DA effect, F(1.864, 26.09) = 2.02, p > 0.05; genotype × coincident DA interaction, F(2, 28) = 0.43, p > 0.05). No post hoc tests were performed due to the absence of a significant interaction effect; graph annotations denote significant main effects only. Inhibition duration was quantified on a trial-by-trial basis, capturing variability in event timing not reflected in group averages displayed in (E). (I) ACh inhibition amplitude in (E) ( n = 7–9 per condition; two-way repeated measures ANOVA: genotype effect, F(1, 14) = 2.42, p > 0.05; coincident DA effect, F(1.370, 19.17) = 3.39, p > 0.05; genotype × coincident DA interaction, F(2, 28) = 2.94, p > 0.05).

Article Snippet: ChrimsonR excitation, GRAB sensor excitation, isosbestic excitation (405 nm), and signal collection were all performed through a single patch cord (MFP_200/220/LWMJ-0.37_1m_FCM-MF1.25(F)_LAF; Doric Lenses), connected to the animal’s implanted optical fiber via an interconnect (ADAL3; Thorlabs), and to the sample (S) port of the Mini Cube.

Techniques: Inhibition, Expressing, Control