16-bit grayscale images of nadh and fad channels for the kidney  (MathWorks Inc)

Journal: Frontiers in Neuroscience

Article Title: Identification of Region-Specific Cytoskeletal and Molecular Alterations in Astrocytes of Mecp2 Deficient Animals

doi: 10.3389/fnins.2022.823060

Figure Lengend Snippet: Mecp2 KO astrocytes in the motor cortex (layer I) exhibit cytoskeletal atrophy in early-symptomatic (P20) as well as in moderate (P40) and severe symptomatic animals (P70). (A) Representative images of brain coronal sections, with the motor cortex (layer I) highlighted in red. (B) Micrographs are representative images of WT and Mecp2 KO astrocytes immunostained for GFAP (green) and DAPI (blue) in the motor cortex (layer I) at P20. Scale bar = 10 μm. (C,D) The graphs show the total length of processes (C) and their number (D) in astrocytes in the motor cortex of WT and Mecp2 KO mice at P20, P40, and P70. Data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by Student’s t -test or Mann–Whitney test in accordance with data distribution. One-way ANOVA indicates no significant alteration in WT and KO astrocytes along time. (E–G) The graphs depict data from Sholl analysis, reporting the number of intersections of astrocyte processes with concentric circles, in the motor cortex of WT and Mecp2 KO mice at P20 (E) , P40 (F) , and P70 (G) . Representative images of reconstructed astrocyte arbors by SNT plugin are reported above each graph. * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA, followed by Sidak’s multiple comparison test. WT and Mecp2 KO astrocytes (P20: n = 36 WT and n = 35 KO; P40: n = 27 WT and n = 44 KO; P70: n = 50 WT and n = 53 KO; n indicates the number of cells) derived from at least three different animals per genotype (P20: N = 4 WT and N = 4 KO; P40: N = 3 WT and N = 5 KO; P70: N = 5 WT and N = 6 KO; N indicates the number of animals).

Article Snippet: For the analysis of GFAP + astrocytes, Z-stack images (212.13 × 212.13 μm , 1,024 × 1,024 pixel resolution, 16-bit grayscale depth) of brain sections were acquired at a Nikon Ti2 Microscope equipped with an A1 + laser scanning confocal system and a Plan Apo λ 60× oil-immersion objective with a step size of 0.5 μm.

Techniques: MANN-WHITNEY, Comparison, Derivative Assay

Journal: Frontiers in Neuroscience

Article Title: Identification of Region-Specific Cytoskeletal and Molecular Alterations in Astrocytes of Mecp2 Deficient Animals

doi: 10.3389/fnins.2022.823060

Figure Lengend Snippet: In the somatosensory cortex (layer I) Mecp2 KO astrocytes show slight cytoskeletal alterations in early-symptomatic mice (P20), that worse in moderate (P40) and severe symptomatic animals (P70). (A) Representative images of brain coronal sections, with the somatosensory cortex (layer I) highlighted in red. (B) Micrographs are representative images of WT and Mecp2 KO astrocytes immunostained for GFAP (green) and DAPI (blue) in the somatosensory cortex (layer I) at P40. Scale bar = 10 μm. (C,D) The graphs show the total length of processes (C) and their number (D) in astrocytes in the somatosensory cortex of WT and Mecp2 KO at P20, P40, and P70. Data are represented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by Student’s t -test or Mann–Whitney test in accordance with data distribution. One-way ANOVA indicates a significant increase in the total length and number of WT and KO astrocyte processes from P20 to P40 ( p < 0.001). (E–G) The graphs depict data from Sholl analysis, reporting the number of intersections of WT and KO astrocyte processes with concentric circles, at P20 (E) , P40 (F) , and P70 (G) . Representative images of reconstructed astrocyte arbors by SNT plugin are reported above each graph. ** p < 0.01, *** p < 0.001 by two-way ANOVA, followed by Sidak’s multiple comparison test. WT and Mecp2 KO astrocytes (P20: n = 36 WT and n = 36 KO; P40: n = 16 WT and n = 35 KO; P70: n = 34 WT and n = 36 KO; n indicates the number of cells) derived from at least three different animals per genotype (P20: N = 4 WT and N = 4 KO; P40: N = 3 WT and N = 5 KO; P70: N = 4 WT and N = 4 KO; N indicates the number of animals).

Article Snippet: For the analysis of GFAP + astrocytes, Z-stack images (212.13 × 212.13 μm , 1,024 × 1,024 pixel resolution, 16-bit grayscale depth) of brain sections were acquired at a Nikon Ti2 Microscope equipped with an A1 + laser scanning confocal system and a Plan Apo λ 60× oil-immersion objective with a step size of 0.5 μm.

Techniques: MANN-WHITNEY, Comparison, Derivative Assay

Journal: Frontiers in Neuroscience

Article Title: Identification of Region-Specific Cytoskeletal and Molecular Alterations in Astrocytes of Mecp2 Deficient Animals

doi: 10.3389/fnins.2022.823060

Figure Lengend Snippet: In the layer I of the somatosensory cortex of symptomatic heterozygous animals, the astrocyte cytoskeleton is affected regardless of Mecp2 expression. (A) Micrographs are representative images of astrocytes in the somatosensory cortex (layer I) of WT and heterozygous brains at P180. WT astrocytes were immunostained for GFAP (green) and DAPI (blue); heterozygous brains were also immunostained for Mecp2 (red) to discriminate between cells expressing the WT ( Mecp2 + ) or null ( Mecp2 – ) allele. Scale bar = 10 μm. (B,C) The graphs show the total length (B) and the number of processes (C) in Mecp2 + (gray) and Mecp2 – (red) astrocytes derived from the somatosensory cortex (layer I) of P90 and P180 heterozygous female mice, compared to astrocytes of WT animals (black). Data are represented as mean ± SEM. ** p < 0.01 by Kruskal–Wallis test, followed by Dunn’s multiple comparison test. (D,E) The graphs depict data from Sholl analysis, reporting the number of intersections of processes of WT (black), Mecp2 + (gray) and Mecp2 – (red) astrocytes with concentric circles, at P90 (D) and P180 (E) . Representative images of reconstructed astrocyte arbors by SNT plugin are reported above each graph. * p < 0.05, # p < 0.001 by two-way ANOVA, followed by Sidak’s multiple comparison test. * and # indicate the comparison between WT and Mecp2 + astrocytes (gray), WT and Mecp2 – astrocytes (red), Mecp2 + and Mecp2 – astrocytes (black). Four animals per genotype were used at each time point ( N = 4). Astrocytes were randomly selected in the acquired field (P90–110: n = 44 WT, n = 46 Mecp2 + , n = 18 Mecp2 – ; P180: n = 49 WT, n = 35 Mecp2 + and n = 21 Mecp2 – ; n indicates the number of cells).

Article Snippet: For the analysis of GFAP + astrocytes, Z-stack images (212.13 × 212.13 μm , 1,024 × 1,024 pixel resolution, 16-bit grayscale depth) of brain sections were acquired at a Nikon Ti2 Microscope equipped with an A1 + laser scanning confocal system and a Plan Apo λ 60× oil-immersion objective with a step size of 0.5 μm.

Techniques: Expressing, Derivative Assay, Comparison

Journal: Frontiers in Neuroscience

Article Title: Identification of Region-Specific Cytoskeletal and Molecular Alterations in Astrocytes of Mecp2 Deficient Animals

doi: 10.3389/fnins.2022.823060

Figure Lengend Snippet: Hippocampal (CA1 pyramidal layer) Mecp2 KO astrocytes manifest signs of cytoskeletal atrophy only in severe symptomatic animals (P70), but not at earlier time points. (A) Representative images of brain coronal sections, with the CA1 of dorsal hippocampus highlighted in red. (B) Micrographs are representative images of WT and Mecp2 KO astrocytes immunostained for GFAP (green) and DAPI (blue) in the CA1 pyramidal layer of the hippocampus at P70. Scale bar = 10 μm. (C,D) The graphs show the total length of processes (C) and their number (D) in Mecp2 KO astrocytes, compared to WT astrocytes, in the CA1 of dorsal hippocampus of P20, P40, and P70 mice. Data are represented as mean ± SEM. *** p < 0.001 by Student’s t -test or Mann–Whitney test in accordance with data distribution. One-way ANOVA indicates a progressive and significant increase in the total length of processes of WT astrocytes (from P20 to P40 p < 0.001; from P40 to P70 p < 0.01), and an arrest of growth at P40 in KO cells (from P20 to P40 p < 0.001). The number of processes increases from P20 to P40 ( p < 0.001) and slightly decreases at P70 in both WT ( p = 0.061) and KO ( p < 0.01) astrocytes. (E–G) The graphs depict data from Sholl analysis, reporting the number of intersections of WT and Mecp2 KO astrocyte processes with concentric circles, at P20 (E) , P40 (F) , and P70 (G) . Representative images of reconstructed astrocyte arbors by SNT plugin are reported above each graph. * p < 0.05, *** p < 0.001 by two-way ANOVA, followed by Sidak’s multiple comparison test. WT and Mecp2 KO astrocytes (P20: n = 36 WT and n = 36 KO; P40: n = 27 WT and n = 45 KO; P70: n = 36 WT and n = 36 KO; n indicates the number of cells) derived from at least three different animals per genotype (P20: N = 4 WT and N = 4 KO; P40: N = 3 WT and N = 5 KO; P70: N = 4 WT and N = 4 KO; N indicates the number of animals).

Article Snippet: For the analysis of GFAP + astrocytes, Z-stack images (212.13 × 212.13 μm , 1,024 × 1,024 pixel resolution, 16-bit grayscale depth) of brain sections were acquired at a Nikon Ti2 Microscope equipped with an A1 + laser scanning confocal system and a Plan Apo λ 60× oil-immersion objective with a step size of 0.5 μm.

Techniques: MANN-WHITNEY, Comparison, Derivative Assay

Journal: Frontiers in Neuroscience

Article Title: Identification of Region-Specific Cytoskeletal and Molecular Alterations in Astrocytes of Mecp2 Deficient Animals

doi: 10.3389/fnins.2022.823060

Figure Lengend Snippet: The genes and the primers used for qRT-PCR are reported.

Article Snippet: For the analysis of GFAP + astrocytes, Z-stack images (212.13 × 212.13 μm , 1,024 × 1,024 pixel resolution, 16-bit grayscale depth) of brain sections were acquired at a Nikon Ti2 Microscope equipped with an A1 + laser scanning confocal system and a Plan Apo λ 60× oil-immersion objective with a step size of 0.5 μm.

Techniques: