anti-rabbit htr2a antibody ra24288  (Neuromics)

Journal: bioRxiv

Article Title: Psilocybin Promotes Cell-Type-Specific Changes in the Orbitofrontal Cortex Revealed by Single-Nucleus RNA-seq

doi: 10.1101/2024.01.07.573163

Figure Lengend Snippet: (A) The sampled for single nucleus RNA sequencing (snRNA-seq) experiment was performed 24 hrs after i.p. injection of psilocybin (Psilocybin) or the vehicle (Saline). (B) Visualization of cell clusters in the OFC with t-distributed stochastic neighbour embedding ( t -SNE). Dots represent individual cells, which were clustered into 3 major groups, 20 subgroups (Saline, n = 12246; Psilocybin, n = 12991; N = 4/group). (C) Dot plot of the expression of cell-type markers in clusters. (D) Violin plot of Htr2a expression in clusters. (E) RNAScope in situ hybridization analysis of Htr2a in layer 5 excitatory ( Slc17a7 +), inhibitory PV+ ( Pvalb +), and inhibitory SST+ ( Sst +) neurons of the OFC in Saline and Psilocybin mice. Top panels, characteristic images of Htr2a mRNA puncta in Slc17a7 +, Pvalb +, and Sst + cells in layer 5 of OFC. Bottom boxplots, the comparisons of the density of Htr2a puncta in the three cell types ( p , Wilcoxon test; Slc17a7 +: Saline, n =52; Psilocybin, n = 55; Pvalb +: Saline, n = 74; Psilocybin, n = 68; Sst +: Saline, n = 50; Psilocybin, n = 52; N = 3/group). (F) Volcano plots of the differential gene expression analysis of the Ex1, Pvalb, and Sst cluster of snRNA-seq data between Psilocybin and Saline mice. (G) Reactome pathway-based gene set enrichment analysis of differential gene expressions in the Ex1 cluster between Psilocybin and Saline mice. (H) Network analysis of genes showing differential expression in Psilocybin mice with top annotated Reactome pathway categories of the Ex1 cluster.

Article Snippet: Primary antibodies used were Rabbit Anti-HTR2A (1:400, Thermo Fisher, Cat# PA5-120747, RRID:AB_2914319), Rabbit Anti-Glutamate receptor 1 (1:150, Millipore, Cat# AB1504, RRID:AB_2113602), Rat Anti-Somatostatin (1:200, Millipore, Cat# MAB354, RRID:AB_2255365), Goat Anti-Parvalbumin (1:2000, Swant, Cat# PVG-213, RRID:AB_2650496), Goat Anti-Iba1 (1:100, Abcam, Cat# ab289874, RRID:AB_2942069), Mouse Anti-Glial Fibrillary Acidic Protein (1:100, Millipore, Cat# MAB360, RRID:AB_11212597), Mouse Anti-NeuN (1:1000, Abcam, Cat# ab104224, RRID:AB_10711040), and Rat Anti-NeuN (1:2000, Abcam, Cat# ab279297).

Techniques: RNA Sequencing Assay, Injection, Saline, Expressing, In Situ Hybridization

Journal: bioRxiv

Article Title: Psilocybin Promotes Cell-Type-Specific Changes in the Orbitofrontal Cortex Revealed by Single-Nucleus RNA-seq

doi: 10.1101/2024.01.07.573163

Figure Lengend Snippet: (A) Representative images of Htr2a mRNA expression in excitatory ( Slc17a7 +), inhibitory PV+ ( Pvalb +), and inhibitory SST+ ( Sst +) neurons of layer 5 in the OFC. Dashed squares indicate the cells shown in Figure 1E . (B) Statistical comparison of Htr2a puncta counts in Slc17a7 +, Pvalb +, and Sst + cells ( p , Wilcoxon test; Slc17a7 +: Saline, n =52; Psilocybin, n = 55; Pvalb +: Saline, n = 74; Psilocybin, n = 68; Sst +: Saline, n = 50; Psilocybin, n = 52; N = 3/group).

Article Snippet: Primary antibodies used were Rabbit Anti-HTR2A (1:400, Thermo Fisher, Cat# PA5-120747, RRID:AB_2914319), Rabbit Anti-Glutamate receptor 1 (1:150, Millipore, Cat# AB1504, RRID:AB_2113602), Rat Anti-Somatostatin (1:200, Millipore, Cat# MAB354, RRID:AB_2255365), Goat Anti-Parvalbumin (1:2000, Swant, Cat# PVG-213, RRID:AB_2650496), Goat Anti-Iba1 (1:100, Abcam, Cat# ab289874, RRID:AB_2942069), Mouse Anti-Glial Fibrillary Acidic Protein (1:100, Millipore, Cat# MAB360, RRID:AB_11212597), Mouse Anti-NeuN (1:1000, Abcam, Cat# ab104224, RRID:AB_10711040), and Rat Anti-NeuN (1:2000, Abcam, Cat# ab279297).

Techniques: Expressing, Comparison, Saline

Journal: bioRxiv

Article Title: Psilocybin Promotes Cell-Type-Specific Changes in the Orbitofrontal Cortex Revealed by Single-Nucleus RNA-seq

doi: 10.1101/2024.01.07.573163

Figure Lengend Snippet: (A) shRNA reduced 5-HT 2A receptor subunit expression in layer 5 pyramidal neurons ( p , Wilcoxon test; Scramble, n = 45; sh- Htr2a , n = 56; N = 3/group). (B) sh- Htr2a blocked psilocybin induced GluR1 down-regulation in layer 5 pyramidal neurons. Psilocybin or saline was i.p. injected 24 hrs before tissue collection (Two-way ANOVA, F (3,156) = 5.04, p = 0.026; post hoc Tukey test: Saline + Scramble vs Psilocybin + Scramble, p = 0.0046; Psilocybin + Scramble vs Psilocybin + sh- Htr2a , p = 0.00049; Saline + sh- Htr2a vs Psilocybin + Scramble, p = 0.00084; Saline + Scramble, n = 39; Saline + sh- Htr2a , n = 33; Psilocybin + Scramble, n = 46; Psilocybin + sh- Htr2a , n = 42; N = 3/group) (C) Cell-type specific knockdown of Htr2a on the anti-depressant effect of psilocybin in rFST. Top panel, experiment timeline, arrowheads indicate timepoints of FSTs. Psilocybin or saline was i.p. injected 24 hrs after the FST on day 5 (Pre-Test). Bottom panel, the comparisons of the difference in immobile times of the last two FSTs, namely FST on day 13 (Test) and FST on day 5 (Pre-Test) (One-way ANOVA, F (3,29) = 10.09, p = 0.0001; post hoc Holm test: Camk2-Scramble + Saline vs Camk2-Scramble + Psilocybin, p = 6.3 x 10 -5 ; Camk2-Scramble + Psilocybin vs Camk2-sh- Htr2a + Psilocybin, p = 0.02; Camk2-Scramble + Saline vs PV-sh- Htr2a + Psilocybin, p = 0.02; Camk2-Scramble + Saline, N = 10; Camk2-Scramble + Psilocybin, N = 10; Camk2-sh- Htr2a + Psilocybin, N= 6; PV-sh- Htr2a + Psilocybin, N = 7).

Article Snippet: Primary antibodies used were Rabbit Anti-HTR2A (1:400, Thermo Fisher, Cat# PA5-120747, RRID:AB_2914319), Rabbit Anti-Glutamate receptor 1 (1:150, Millipore, Cat# AB1504, RRID:AB_2113602), Rat Anti-Somatostatin (1:200, Millipore, Cat# MAB354, RRID:AB_2255365), Goat Anti-Parvalbumin (1:2000, Swant, Cat# PVG-213, RRID:AB_2650496), Goat Anti-Iba1 (1:100, Abcam, Cat# ab289874, RRID:AB_2942069), Mouse Anti-Glial Fibrillary Acidic Protein (1:100, Millipore, Cat# MAB360, RRID:AB_11212597), Mouse Anti-NeuN (1:1000, Abcam, Cat# ab104224, RRID:AB_10711040), and Rat Anti-NeuN (1:2000, Abcam, Cat# ab279297).

Techniques: shRNA, Expressing, Saline, Injection

Journal: bioRxiv

Article Title: Psilocybin Promotes Cell-Type-Specific Changes in the Orbitofrontal Cortex Revealed by Single-Nucleus RNA-seq

doi: 10.1101/2024.01.07.573163

Figure Lengend Snippet: (A -C) Representative viral infection in the OFC 2 weeks after viruses injections. (D) The time course of immobile time in FSTs, related to . Psilocybin or saline was i.p. injected 24 hrs after the FST on day 5 (Test Day 1: One-way ANOVA, F (3,29) = 1.11, p = 0.36; Test Day 5: One-way ANOVA, F (3,29) = 0.44, p = 0.72; Saline, N = 10; Psilocybin, N = 10; Camk2-sh- Htr2a , N= 6; PV-sh- Htr2a , N = 7).

Article Snippet: Primary antibodies used were Rabbit Anti-HTR2A (1:400, Thermo Fisher, Cat# PA5-120747, RRID:AB_2914319), Rabbit Anti-Glutamate receptor 1 (1:150, Millipore, Cat# AB1504, RRID:AB_2113602), Rat Anti-Somatostatin (1:200, Millipore, Cat# MAB354, RRID:AB_2255365), Goat Anti-Parvalbumin (1:2000, Swant, Cat# PVG-213, RRID:AB_2650496), Goat Anti-Iba1 (1:100, Abcam, Cat# ab289874, RRID:AB_2942069), Mouse Anti-Glial Fibrillary Acidic Protein (1:100, Millipore, Cat# MAB360, RRID:AB_11212597), Mouse Anti-NeuN (1:1000, Abcam, Cat# ab104224, RRID:AB_10711040), and Rat Anti-NeuN (1:2000, Abcam, Cat# ab279297).

Techniques: Infection, Saline, Injection