goat anti substance p  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: Involvement of prelimbic cortex neurons and related circuits in the acquisition of cooperative learning by pairs of rats

doi: 10.1101/2022.01.13.476162

Figure Lengend Snippet: A, Immunofluorescence procedure. Animals (n = 40) were randomly allocated into 4 groups of 10 rats: control group (blue), group 1 (purple), group 2 (pink), and group 3 (dark red). All animals underwent a battery of behavioral tests before the cooperation experiment: open field test (OFT), elevated plus maze (EPM), water competition test (WCT), and food competition test (FCT). After that, groups 1, 2, and 3 started the cooperation experiment. The brains of each group were collected after reaching the established key moments (indicated by syringe icons in the image): brains of group 1 were collected after reaching the criterion for the individual phase (i.e., ≥ 60 pellets for 2 consecutive days), brains in group 2 after reaching the criterion for the cooperative phase (i.e., ≥ 40 pellets for 2 consecutive days), and brains of group 3 at the end of the experiment. Animals in the control group remained in their home cages during the cooperation experiment and were handled and weighed daily as the remaining animals. After the cooperation experiment, all the brains were cryoprotected, and coronal sections of the PrL cortex were labeled for c-FOS, DAPI, and selected antibodies against GABAergic and dopaminergic neurons (D1- and D2-containing cells). B-F , Cumulative records showing the acquisition curves for individual and cooperative phases. 38 male rats were randomly allocated into 4 groups (group 1 (individual phase, represented in purple), group 2 (cooperative phase, represented in pink), group 3 (whole experiment, represented in red), and the control group. All groups except the control performed the cooperation experiment. B , Group 1 learned to individually climb and sit on the platform to get a pellet of food, showing a steep slope between the third and the last day of training: average ± SD slope for group 1, 76.801 ± 22.46. The cumulative line stops at the day that rats reached the criterion and were perfused and euthanized. C , D , Groups 2 and 3 performed the individual phase completely and all the animals reached the criterion, showing a steep slope from day 3 to 10 (group 2, slope = 77.03 ± 26.01; and group 3, slope = 94.24 ± 27.45). E , F , After learning the individual task, groups 2 and 3 were placed again in the double Skinner box for phase II (cooperative). This time, they had to coordinate their behavior (climbing and staying together on the platform for > 500 ms) to get a pellet of food. All rats learned the task successfully, showing a steep slope from session 3 to their last session (group 2, slope = 87.82 ± 17.31; group 3, slope = 82.45 ± 14.04).

Article Snippet: After that, sections were incubated for 40 h at 4 °C with rabbit anti-c-FOS (Synaptic Systems, 226003) or goat anti-c-FOS (Santa Cruz, sc-52-G) and one of the following primary antibodies: mouse anti-GAD67 for staining GABAergic cells (Merck & Co., MAB5406), goat anti-Substance P for staining D1-containing cells (Santa Cruz, sc-9758), and rabbit anti-Met Enkephalin for staining D2-containing cells, (Abcam, ab22620).

Techniques: Immunofluorescence, Labeling

Journal: bioRxiv

Article Title: Involvement of prelimbic cortex neurons and related circuits in the acquisition of cooperative learning by pairs of rats

doi: 10.1101/2022.01.13.476162

Figure Lengend Snippet: A , C , Coronal brain sections from animals in groups 2 and 3 labeled for c-FOS (red channel), DAPI (blue channel), and the selected antibodies against the main cell types of the PrL. Goat anti-Substance P for staining D1-containing cells (green channel) and rabbit anti-Enkephalin for staining D2-containing cells (gray channel). The photomicrographs corresponding to Group 2, in which rats were trained to cooperate until reaching the criterion for the cooperation phase, are shown in A, and the photomicrographs in C correspond to group 3, which performed the whole cooperative phase (10 sessions). B , D , Percentage of c-FOS activation in each group. B , D1-containing cells were significantly more active in leaders than in followers during the cooperation phase (One-way ANOVA, F = 16.95, p = 0.007). The activation of D2-containing cells was lower than that of D1 cells, and the highest activation was also observed in the leader rats from group 2. However, the difference between leaders and followers was not significant (One-way ANOVA, F = 1.63, p = 0.24). Leader rats from group 3 ( C ), which completed the 10 days of the task, also showed higher activation for D1 and D2 cells, but the difference was not significant (One-way ANOVA, F = 1.44, p = 0.29).

Article Snippet: After that, sections were incubated for 40 h at 4 °C with rabbit anti-c-FOS (Synaptic Systems, 226003) or goat anti-c-FOS (Santa Cruz, sc-52-G) and one of the following primary antibodies: mouse anti-GAD67 for staining GABAergic cells (Merck & Co., MAB5406), goat anti-Substance P for staining D1-containing cells (Santa Cruz, sc-9758), and rabbit anti-Met Enkephalin for staining D2-containing cells, (Abcam, ab22620).

Techniques: Labeling, Staining, Activation Assay