Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Morphine-withdrawal aversive memories and their extinction modulate H4K5 acetylation and Brd4 activation in the rat hippocampus and basolateral amygdala.

doi: 10.1016/j.biopha.2023.115055

Figure Lengend Snippet: Fig. 2. (A) Schematic illustrations of coronal sections showing the BLA and the hippocampus (DG, CA1 and CA3) (modified from Paxinos and Watson 2007). Quantitative H4K5ac IR and p-Brd4 IR in the BLA (B, F), DG (C, G), CA1 (D, H) and CA3 (E, I) after conditioned place aversion (CPA) paradigm and after the post- extinction (Ext) CPA test. *P < 0.05, * *P < 0.001, * **P < 0.0001 versus CPA-PLA group (Student’s t test; for details, see Table S2). +P < 0.05 versus MOR-Ext (Kruskal-Wallis ANOVA with Dunn’s post-hoc analysis; n = 4–5 for each group). (J-I’) Representative confocal microphotographs showing coronal section of rats immunostained in: green (H4K5ac) and red (p-Brd4). Scale bar, 25 µm.

Article Snippet: After this first phase, sections were washed and reblocked with BSA solution and were incubated with rabbit polyclonal biotin-conjugated anti H4K5ac (NB21–2024B; 1:1000, Novus Biologicals, Centennial, CO, USA) for 24 h at 4 oC with gentle agitation.

Techniques: Modification

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Morphine-withdrawal aversive memories and their extinction modulate H4K5 acetylation and Brd4 activation in the rat hippocampus and basolateral amygdala.

doi: 10.1016/j.biopha.2023.115055

Figure Lengend Snippet: Fig. 3. Number of H4K5ac- (B) and p-Brd4-positive neurons (C), and percentage of p-BRD4 expressing H4K5ac (D) in the BLA did not change after the CPA test nor after the extinction test (for details, see table S3). The analysed region within the BLA is schematically illustrated in (A). Representative confocal images in the BLA of H4K5ac (green; H-N), p-Brd4 (red; H’-N’) and DAPI (nuclear stain, blue; H’’-N’’). Merged images are shown in (H’’’-N’’’; H4K5ac/p-Brd4). Colocalization is shown by yellow/orange neurons in the nucleus of merged images of H4K5ac/p-Brd4. Scale bars, 30 µm. (I-O, scale bars 30 µm; I’-O’, scale bars 5 µm) Three-dimensional reconstructions of nuclei expressing H4K5ac/p-Brd4 in the BLA after different treatments. (E-G) Quantitative colocalization was performed, and several colocali zation parameters were determined [Pearson’s coefficient, Manders’ coefficient 1-the fraction of objects in channel A (H4K5ac) colocalized with objects in channel B (p-Brd4) and Manders’ coefficient 2-vice versa]. Results are expressed as mean ± SEM. N = 5 animals per group.

Article Snippet: After this first phase, sections were washed and reblocked with BSA solution and were incubated with rabbit polyclonal biotin-conjugated anti H4K5ac (NB21–2024B; 1:1000, Novus Biologicals, Centennial, CO, USA) for 24 h at 4 oC with gentle agitation.

Techniques: Expressing, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Morphine-withdrawal aversive memories and their extinction modulate H4K5 acetylation and Brd4 activation in the rat hippocampus and basolateral amygdala.

doi: 10.1016/j.biopha.2023.115055

Figure Lengend Snippet: Fig. 4. Number of H4K5ac- (B) and p-Brd4-positive neurons (C), and percentage of p-BRD4 expressing H4K5 (D) in the DG after the CPA test and after the extinction test. Results are shown as mean ▫ SEM. *P < 0.05 versus the PLA-CPA group (for details, see table S4). The analysed region within the DG is schematically illustrated in (A). Representative confocal images in the DG of H4K5ac (green; H-N), p-Brd4 (red; H’-N’) and DAPI (nuclear stain, blue; H’’-N’’). Merged images are shown in (H’’’-N’’’; H4K5ac/p-Brd4). Colocalization is shown by yellow/orange neurons in the nucleus of merged images of H4K5/p-Brd4. Scale bars, 30 µm. (I-O, scale bars 30 µm; I’-O’, scale bars 5 µm) Three-dimensional reconstructions of nuclei expressing H4K5ac/p-Brd4 in the BLA after different treatments. (E-G) Quantitative colocalization was performed, and several colocalization parameters were determined [Pearson’s coefficient, Manders’ coefficient 1-the fraction of objects in channel A (H4K5ac) colocalized with objects in channel B (p-Brd4) and Manders’ coefficient 2-vice versa]. Results are expressed as mean ± SEM. N = 5 animals per group.

Article Snippet: After this first phase, sections were washed and reblocked with BSA solution and were incubated with rabbit polyclonal biotin-conjugated anti H4K5ac (NB21–2024B; 1:1000, Novus Biologicals, Centennial, CO, USA) for 24 h at 4 oC with gentle agitation.

Techniques: Expressing, Staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Morphine-withdrawal aversive memories and their extinction modulate H4K5 acetylation and Brd4 activation in the rat hippocampus and basolateral amygdala.

doi: 10.1016/j.biopha.2023.115055

Figure Lengend Snippet: Fig. 5. Number of H4K5ac- and p-Brd4-positive neurons, and percentage of p-Brd4 expressing H4K5ac in the CA1 (B-D) and CA3 (P-R) after the CPA test and after the extinction test. Results are shown as mean ± SEM. N = 4–5 per each group. *P < 0.05 versus the PLA-CPA-group (Kruskal-Wallis ANOVA with Dunn’s post-hoc analysis). The analysed regions within the CA1 and CA3 are schematically illustrated in (A). Representative confocal images in the CA1 of H4K5ac (green; H-N), p- Brd4 (red; H’-N’) and DAPI (nuclear stain, blue; H’’-N’’) and in the CA3 (V-AB for H4K5, green; V’-AB’ for p-Brd4, red; and V’’-AB’’ for DAPI, blue). Merged images are shown in H’’’-N’’’ for CA1 and V’’’-AB’’’ for CA3; H4K5ac/p-Brd4). Colocalization is shown by yellow/orange neurons in the nucleus of merged images of H4K5ac/p-Brd4. Scale bars, 30 µm. (I-O for CA1 and W-AC for CA3, scale bars 30 µm; I’-O’ for CA1 and W’-AC’ for CA3, scale bars 5 µm) Three-dimensional re constructions of nuclei expressing H4K5ac/p-Brd4 in the CA1 and CA3 after different treatments. (E-G; S-U) Quantitative colocalization was performed, and several colocalization parameters were determined [Pearson’s coefficient, Manders’ coefficient 1-the fraction of objects in channel A (H4K5ac) colocalized with objects in channel B (p-Brd4) and Manders’ coefficient 2-vice versa]. Results are expressed as mean ± SEM. N = 5 animals per group.

Article Snippet: After this first phase, sections were washed and reblocked with BSA solution and were incubated with rabbit polyclonal biotin-conjugated anti H4K5ac (NB21–2024B; 1:1000, Novus Biologicals, Centennial, CO, USA) for 24 h at 4 oC with gentle agitation.

Techniques: Expressing, Staining