grin lens proviewtm lens probe  (Inscopix Inc)

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , The strategy to monitor rACC→Pn neuron activity. Scale bar, 2 mm. b , Maximal projection of a Ca 2+ video with 82 rACC→Pn neurons. Scale bar, 50 μm. c , z -scored Ca 2+ activity for the neurons numbered in b . d , rACC→Pn neuron activity during the first border crossing. e , f , rACC→Pn neuron activity averaged for individual neurons ( e ; F 2,612 = 5.16, P = 0.006; n = 205) and individual mice ( f ; F 2,15 = 15.6, P = 0.0002; n = 6). g , rACC→Pn neuron activity during border crossings on the post-test day. h , i , Averaged rACC→Pn neuron activity for individual neurons ( h ; F 2,954 = 10.13, P = 4 × 10 −5 ; n = 320) and individual mice ( i ; F 2,15 = 16.8, P = 0.0002; n = 6). j , The latency preceding the first border crossing against the averaged signal of rACC→Pn neurons for each mouse, with the linear regression fit. k , Recording (rec.) configuration of rACC→Pn neurons. IN, local interneurons; PN, pyramidal neurons; II/III, layers II and III of ACC. l , EPSCs evoked at different holding potentials. The dot represents the peak AMPAR EPSC; the dashed line indicates the NMDAR EPSC amplitude. m , The AMPAR/NMDAR ratio ( P = 0.03). n = 11 (Ctrl) and 10 (Cond.) neurons. n , TBS-induced EPSC amplitude changes. Inset: averaged EPSCs before and after TBS. n = 6 neurons per group. o , Recordings of isolated EPSCs and IPSCs. The arrows indicate the onset of electrical stimulation to onset of EPSCs and IPSCs. p , The EPSC–IPSC conductance ratio ( P = 0.005). q , The EPSC–IPSC delay ( P = 8 × 10 −5 ). n = 8 (Ctrl) and 10 (Cond.) neurons. Statistical analysis was performed using one-way ANOVA with Tukey post hoc test ( e , f , h and i ), two-sided Wilcoxon rank-sum tests ( m , p and q ) and Pearson’s two-sided correlation tests ( l ). For d and g , neurons are ordered by mean Ca 2+ activity for each condition. For the box plots, the centre lines show the median values, the box limits show the quartiles, and the whiskers show the most extreme datapoints ≤interquartile range from the box edges. For n , data are mean ± s.e.m. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Activity Assay, Isolation

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , Cross-day alignment of rACC→Pn neurons across Pre, Cond, and Post phases of PAC using TRACKER. Neurons matched across 3 days are outlined in blue. Scale bar, 50 μm. b , Boxplot of the number of detected rACC→Pn neurons during Pre (grey), Cond (green), and Post (blue) phases, and the number of cross-day-aligned neurons (red). n = 6 mice. c , Z-scored activity traces (coloured traces) for the 10 neurons in (a) . Raster traces show the binarized patterns of activity for each neuron. d , Mice with intracranial virus injection, GRIN lens implantation, and miniature microscope mounting (Mini) showed no difference in total walking distance and average movement speed versus mice without these manipulations (Ctrl) during PAC. n = 17 and 6 mice in Ctrl and Mini groups. e , Average Ca 2+ activity of rACC→Pn neurons during first border crossing as a function of time in Pre (red), day 5 (green), and day 6 (blue). Note that their activity increased progressively during the conditioning phase of PAC. f , Line graph showing the progressively increased activity of rACC→Pn neurons during the conditioning phase of PAC. n = 6 mice in each group. g , Venn diagram showing cross-day-aligned rACC→Pn neurons that show increased activity in pairwise comparisons between Pre, Cond, and Post. h , Boxplots of the proportion of cross-day-aligned rACC→Pn neurons that show increased activity in pairwise comparisons between Pre, Cond, and Post. n = 6 mice. i , Discriminability index (d’) calculated between first crossing and crossing back for Ca 2+ traces, averaged for individual neurons (left; P = 2 × 10 −7 ; n = 233) and individual mice (right; P = 0.03; n = 6). j , Average firing rate of rACC→Pn neurons during first border crossing in Pre (cyan), Cond (green) and Post (blue; one-way ANOVA, Tukey post-hoc test, F (2,15) = 5, P = 0.022; n = 6 mice). k , Average firing rate as a function of time of rACC→Pn neurons during first border crossing in Pre (cyan), Cond (green) and Post (blue). l , Average firing rate of rACC→Pn neurons during first border crossing (red), first crossing back (green) and last border crossing (blue) on post-test day (one-way ANOVA, Tukey post-hoc test, F (2,15) = 16.8, P = 0.001; n = 6 mice). m , Discriminability index (d’) calculated between first crossing and crossing back for firing rate, averaged for individual neurons (left; two-sided Wilcoxon matched-pairs signed-rank test, P = 0.004; n = 233 neurons) and for individual mice (right; two-sided Wilcoxon matched-pairs signed-rank test, P = 0.07; n = 6). n , Control for ( i , left) with randomized border crossing time. o , Control for Fig. with randomized border crossing time. p , Control for ( i , right) with randomized border crossing time. q , Control for (j) with randomized border crossing time. r , Control for ( m , left) with randomized border crossing time. s , Control for ( m , right) with randomized border crossing time. t , Boxplots of the correlation coefficient between the activity of rACC→Pn neurons and the velocity of mice during PAC with real neuronal activity trace (red) or randomly shuffled traces (green). n = 6 mice in each group. u , Scatterplot depicting the relationship between the Ca 2+ activity of rACC→Pn neurons and the velocity of mice, specifically during periods of high neuronal activity in the pre-test. Shaded area represents the 95% confidence interval. v , Similar to (u) , but specifically for periods during which mice display high moving velocity in the pre-test. Shaded area represents the 95% confidence interval. w , Average Ca 2+ activity of rACC→Pn neurons over time during the first border crossing, illustrated in a longer time scale for Pre (cyan), Cond (green), and Post (blue). Note that the activity of rACC→Pn neurons decreases after reaching chamber 2 in Cond and Post. n = 6 in each group. Shaded area represents mean ± SEM. Pearson's two-sided correlation test in (u, v) . In boxplots, horizontal lines represent median; boxes, quartiles; whiskers, most extreme data points ≤ interquartile range from box edges; and single points, data from individual cells or mice. Data in (f) , and shaded area in (k) and (w) are mean ± SEM.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Activity Assay, Virus, Injection, Microscopy, Control

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , Strategy and experimental timeline to record IT neuron activity in the rACC during PAC using in vivo Ca 2+ imaging. b , Maximal projection of a Ca 2+ video with 37 IT neurons. Scale bar, 50 μm. c , Z-scored activity traces (coloured traces) of the 10 example IT neurons outlined in (b). d , Boxplot of the number of detected IT neurons during Pre (grey), Cond (green), and Post (blue) phases, and the number of cross-day-aligned neurons (red; n = 3 mice). e , Activity of cross-day-aligned IT neurons (n = 133 cells from 3 mice) during first border crossing in Pre (left), Cond (middle) and Post (right) phases of PAC. Neurons are ordered according to their mean Ca 2+ activity for each day. f , Averaged activity of IT neurons during first border crossing at the level of individual neurons in Pre (grey), Cond (green) and Post (blue) phases of PAC (one-way ANOVA, Tukey post-hoc test, F (2,396) = 2.19, P = 0.113; n = 133 neurons). g , Similar to (f), averaged for individual mice. h , Fisher information calculated between first crossing and crossing back for Ca 2+ traces, averaged for individual neurons (two-sided Wilcoxon matched-pairs signed-rank test, P = 0.03; n = 133 neurons). i , Similar to (h), averaged for individual mice. n = 3 mice. j , Activity of cross-day-aligned IT neurons (n = 218 from 3 mice) during first border crossing (left), first crossing back (middle), and last border crossing (right) on post-test day. Neurons are ordered according to their mean Ca 2+ activity for each condition. k , Averaged activity of IT neurons during first border crossing (red), first crossing back (green), and last border crossing (blue) at the level of individual cells on the post-test day (one-way ANOVA, Tukey post-hoc test, F (2,651) = 5.41, P = 0.004; n = 218 neurons). l , Similar to (k) , for individual mice. n = 3 mice. m , Average firing rate of IT neurons during first border crossing in Pre (cyan), Cond (green), and Post (blue) phases of PAC. n = 3 mice. n , Average firing rate of rACC→Pn neurons during first border crossing (red), first crossing back (green), and last crossing (blue) on the post-test day. n = 3 mice. In boxplots, horizontal lines represent median; boxes, quartiles; whiskers, most extreme data points ≤ interquartile range from box edges; and single points, data from individual neurons or mice.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Activity Assay, In Vivo, Imaging

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , Strategy to measure innocuous mechanical, noxious mechanical, or noxious heat responses while monitoring the Ca 2+ activity of rACC→Pn neurons. b , Average silhouette width plotted for cluster numbers k from 1 to 10. Dashed line indicates the number of clusters corresponding to the maximal silhouette width. c , Averaged response (10 trials) of individual neurons upon each stimulation. Dashed lines (red) indicate the time point of stimulation (n = 335, 238, and 284 neurons for innocuous mechanical, noxious mechanical and noxious heat, respectively). Red and blue bars represent the cluster to which each neuron belongs (cluster 1 or cluster 2, respectively). d , Average activity of neurons in response to innocuous mechanical, noxious mechanical and noxious heat for cluster 1 (left) and 2 (right). e , Average activity of individual neurons in response to innocuous mechanical, noxious mechanical and noxious heat in cluster 1 (left; one-way ANOVA, Tukey post-hoc test, F (2,286) = 20.26, P = 1.1 × 10 −8 ; n = 124, 92, and 73 neurons for innocuous mechanical, noxious mechanical and noxious heat, respectively) and 2 (right; one-way ANOVA, Tukey post-hoc test, F (2,565) = 16.73, P = 5.1 × 10 −6 ; n = 211, 146, and 211 neurons for innocuous mechanical, noxious mechanical and noxious heat, respectively). Note that innocuous mechanical stimuli induced an increase in the activity of neurons in cluster 1, but a decrease in the activity of neurons in cluster 2. f , Average Ca 2+ activity of rACC→Pn neurons during innocuous mechanical (red), noxious mechanical (green), and noxious heat (blue; n = 5 mice). g , Similar to (c) , with cross-session-aligned neurons (n = 118 neurons). h , Similar to (d) , with cross-session-aligned neurons. i , Similar to (e) , with cross-session-aligned neurons (cluster 1; left; one-way ANOVA, Tukey post-hoc test, F (2,67) = 3.9, P = 0.02; n = 25, 20, and 25 neurons for innocuous mechanical, noxious mechanical and noxious heat; cluster 2; right; one-way ANOVA, Tukey post-hoc test, F (2,281) = 3.2, P = 0.04; n = 93, 98, and 93 neurons for innocuous mechanical, noxious mechanical and noxious heat). j , Similar to (f) , with cross-session-aligned neurons. k , Heatmap of rACC→Pn neuron activity relative to licking (left) and rearing (right) behaviours in the post-test of PAC. l , Average Ca 2+ activity of rACC→Pn neurons during licking (red) and rearing (green). n = 6 mice. In boxplots, horizontal lines represent median; boxes, quartiles; whiskers, most extreme data points ≤ interquartile range from box edges; and single points, data from individual cells or mice. Shaded area in (d, f, h, j, l) represents mean ± SEM.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Activity Assay

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , Strategy to label Pn-projecting rACC neurons for electrophysiological recording. b , Time mice spent in chamber 2 during days 3–6 of PAC. n = 7, 8 in Ctrl and Cond groups. c , Boxplots of the resting membrane potential (RMP; left; P = 0.37) and the input resistance (right; P = 0.44) of rACC→Pn neurons from Ctrl and Cond mice. n = 14, 16 neurons in Ctrl and Cond groups, respectively. d , Boxplots of the peak amplitude (left; P = 0.85) and half-duration of the action potentials (right; P = 0.39). n = 14, 16 neurons in Ctrl and Cond groups, respectively. e , Action potential firing pattern of rACC→Pn neurons from littermate control (cyan) and PAC-conditioned mice (red). f , Action potential firing frequency evoked by different levels of injected current. n = 14 neurons/group. g , Percentage of rACC→Pn neurons displaying different numbers of spikelets in the first action potential. n = 14 neurons in Ctrl and 16 in Cond. h , Traces of the action potential firing pattern evoked by 1-s current injection (black, bottom) in tdTomato-negative (non rACC→Pn) neurons from Ctrl (green, top) and Cond (purple, middle) mice. i , Plot of the action potential firing frequency evoked by different levels of injected current (two-way ANOVA, Tukey post-hoc test, F (1,255) = 10.61, P = 0.001; n = 11, 22 neurons in Ctrl and Cond groups, respectively). j , Example traces of sEPSCs in a rACC→Pn neuron holding at −70 mV from a Ctrl mouse (cyan, top) and a Cond mouse (red, bottom). k , Cumulative histograms of sEPSC frequency from Ctrl (cyan) and Cond (red) mice. Inset: boxplot of sEPSC frequency ( P = 0.56; n = 11 neurons in each group). l , Cumulative histograms of sEPSC amplitude from Ctrl (cyan) and Cond (red) mice. Inset: boxplot of the sEPSC peak amplitude ( P = 0.028; n = 11 neurons in each group). m , Example traces of two EPSCs evoked at 50-ms intervals from Ctrl (cyan, top) and Cond (red, bottom) mice. n , Paired-pulse ratio as a function of Δt (20, 50, 100, 200, 500 ms) between two stimuli (two-way ANOVA, Tukey post-hoc test, F (1,87) = 0.37, P = 0.54) of the inputs to rACC→Pn neurons from Ctrl (cyan) and Cond (red) mice. n = 8, 11 in Ctrl and Cond groups. o , Recordings of mixed EPSCs and IPSCs while holding rACC→Pn neurons at −30 mV from Ctrl (cyan, top) and Cond (red, bottom) mice. Because the holding potential is between the reversal potentials for excitatory and inhibitory events, EPSCs are inwardly directed and IPSCs outwardly directed. p , Strategy to express excitatory opsin (ChR2) in PV + interneurons and label the Pn-projecting rACC neurons for electrophysiological recording. q , Example trace of the action potential firing from a PV + interneuron in the rACC (top) evoked by current injection (bottom). r , Whole-cell recording configuration to analyse the feedforward inhibition from PV + interneurons to rACC→Pn neurons. A blue light (494 nm, 1 ms) was given to evoke neurotransmitter release from PV + interneurons. s , Example trace showing a light-evoked IPSC (blocked by 10 μM SR-95531) in one rACC→Pn neuron from a control mouse. Blue bar indicates the time point of light stimulation. t , Boxplots of 20–80% rise time (left; P = 0.96) and half-duration (right; P = 0.65) of IPSCs from Ctrl (cyan) and Cond (red) mice. n = 8 neurons in each group. u , Paired-pulse ratio as a function of Δt between two light stimulations (20, 50, 100, 200, 500 ms) of the inhibitory inputs from PV + interneurons to rACC→Pn neurons (two-way ANOVA, Tukey post-hoc test, F (1,65) = 0.018, P = 0.89) from Ctrl (cyan) and Cond (red) mice. n = 7 for each group. v , Light-evoked individual IPSCs (grey), and average IPSC (cyan or red) from Ctrl (top) and Cond (bottom) mice. Blue bar indicates the time point of light stimulation and dashed line indicates the IPSC onset of the neuron from the Ctrl group. Inset: average IPSCs at expanded time scale. w , Boxplots of the amplitude (left; P = 0.02) and latency (right; onset to onset; P = 0.01) of light-evoked IPSCs from Ctrl (cyan) and Cond (red) mice. n = 8 cells in each group. Two-sided Wilcoxon rank-sum test was used in (c) , (d) , (i) , (j) , (f) and (w) . In boxplots, horizontal lines represent median; boxes, quartiles; whiskers, most extreme data points ≤ interquartile range from box edges; and single points, data from individual cells. Data in (b, f, i, n, u) are mean ± SEM.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Membrane, Control, Injection, Inhibition

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , The strategy and experimental timeline to optogenetically manipulate the activity of the rACC→Pn pathway. Scale bar, 2 mm. b , The latency preceding first paw licking (left; P = 0.005), rearing (middle; P = 0.04) and jumping (right; P = 0.009) during the post-test. n = 10 mice per group. c , The strategy to measure thermal pain using a hot plate while optogenetically activating or inhibiting the rACC→Pn pathway. d , The latency preceding paw withdrawal on a 48 °C plate (left; F 2,26 = 10.66, P < 0.001) or 52 °C plate (right; F 2,26 = 7.38, P = 0.003). n = 10 (eYFP control), 10 (NpHR) and 9 (ChR2) mice. e , The strategy to measure the mechanical pain threshold with von Frey filaments while optogenetically activating or inhibiting the rACC→Pn pathway. f , Quantification of changes in paw withdrawal frequency in response to six different von Frey filaments induced by optogenetic manipulation of the rACC→Pn pathway ( F 2,156 = 62.965, P = 2 × 10 −16 ). n = 10 (eYFP control), 10 (NpHR) and 9 (ChR2) mice. g , The pain threshold of mice with or without light stimulation ( F 2,26 = 25.98, P = 1.3 × 10 −9 ). n = 10 (eYFP control), 10 (NpHR) and 9 (ChR2) mice. Statistical analysis was performed using two-sided Wilcoxon rank-sum tests ( b ), one-way ANOVA with Tukey post hoc test ( d ) and two-way ANOVA with Tukey post hoc test ( f and g ). For the box plots, the centre lines show the median values, the box limits show the quartiles, and the whiskers show the most extreme datapoints ≤interquartile range from the box edges. For f , data are mean ± s.e.m. * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Activity Assay, Control

Journal: Nature

Article Title: Neural circuit basis of placebo pain relief

doi: 10.1038/s41586-024-07816-z

Figure Lengend Snippet: a , Strategy (top, left) and timeline (bottom, left) to optogenetically activate the rACC→Pn pathway during PAC. Illustration of bilateral implantation of cannula in the Pn (right). Dashed lines indicate cannula location. Scale bar, 500 μm. b , Timeline of the PAC assay, integrating optogenetics. c , Boxplots of the latency preceding first paw licking (two-sided Wilcoxon rank-sum test, P = 0.03; left), rearing (two-sided Wilcoxon rank-sum test, P = 0.24; middle), and jumping (two-sided Wilcoxon rank-sum test, P = 0.51; right). n = 10 in eYFP and 9 in ChR2 groups. d , Boxplot of the change in falling latency of mice during the rotarod test with and without photomanipulation of the rACC→Pn pathway (one-way ANOVA, Tukey post-hoc test, F (2,26) = 1.43, P = 0.25). n = 10 in eYFP, 10 in NpHR, and 9 in ChR2 groups. e , Quantification of the change in paw withdrawal frequency in response to 6 different von Frey filaments during optogenetic manipulation of the rACC→Pn pathway (two-way ANOVA, Tukey post-hoc test, F (2,156) = 55.18, P = 2 × 10 −16 ). n = 10 in eYFP control, 10 in NpHR, and 9 in ChR2 groups. *P < 0.05 and ***P < 0.001. In boxplots, horizontal lines represent median; boxes, quartiles; whiskers, most extreme data points ≤ interquartile range from box edges; and single points, data from individual cells or mice. Data in (d) are mean ± SEM.

Article Snippet: For imaging rACC→Pn neurons, a naked 1.0-mm-diameter gradient refractive index (GRIN) lens probe (1050-004598, Inscopix) was lowered into the implanted cannula using forceps.

Techniques: Optogenetics, Control

Journal: Cell Metabolism

Article Title: Complementary lateral hypothalamic populations resist hunger pressure to balance nutritional and social needs

doi: 10.1016/j.cmet.2023.02.008

Figure Lengend Snippet:

Article Snippet: GRIN lens probe, 0.6 x 7.3 mm , Inscopix , Cat#1050-002208.

Techniques: Virus, Plasmid Preparation, Recombinant, Software, Imaging, Optogenetics