Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: PK profiles of epoD, epoB and ixabepilone following subcutaneous or i.p. injection . ( A ) Timeline of epothilone injections and tissue collection. ( B–D ) Concentration profiles of epothilones in plasma, brain and spinal cord tissue quantified by LC–MS. Created with BioRender.com. Plotted data are the mean drug concentration ±SEM. * P < 0.05 by two-way ANOVA and Bonferroni post hoc (asterisks compares the epoB groups). n = 3 animals per group and time point.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: Injection, Concentration Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: Neither epothilones nor rehabilitation confers neuroprotection . ( A ) Scheme of the study design displaying the timing of injury, rehabilitation and behavioural tests. ( B ) Scheme depicting the location of the contusive injury and what regions of the spinal cord were sectioned for IHC. Created with BioRender.com. ( C ) Representative fluorescence images of GFAP-labelled spinal cord sections at the lesion epicentre or −3600 µm rostral demarcating the lesion area and spared tissue (orange dashed lines; red dashed lines show the tissue border; control animal; pixel values inverted; scale bar = 500 µm). ( D and E ) Quantification of lesion spared tissue and lesion size throughout the lesioned spinal cord determined by GFAP lesion demarcation (control n = 16; epoD n = 18; epoB n = 11; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 12). ( F ) Quantification of the number of neurons throughout the lesioned spinal cord determined by counting NeuN immunolabelling (control n = 13; epoD n = 13; epoB n = 12; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 11). ( G ) Representative fluorescence images of NeuN-labelled spinal cord sections at the lesion epicentre or −3600 µm rostral (control animal; pixel values inverted; scale bar = 500 µm). Plotted data are the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 by two-way ANOVA and Bonferroni post hoc .

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: Fluorescence

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: Epothilone B reduces fibrotic scar components after T10 175 kdyn contusion injury . ( A ) Representative fluorescent images of laminin throughout the lesioned spinal cord for the given groups (pixel values inverted; scale bar = 1 mm). ( B ) Quantification of laminin staining area within every 600 µm regions of spinal cord tissue (control n = 16; epoD n = 18; epoB n = 12; rehabilitation n = 11; epoD + rehabilitation n = 12; epoB + rehabilitation n = 11). ( C ) Representative fluorescent images of fibronectin throughout the lesioned spinal cord for the given groups (pixel values inverted; scale bar = 1 mm). ( D ) Quantification of fibronectin staining area within every 600 µm regions of spinal cord tissue. ( E ) Area of fibronectin staining in regions rostral (−4200 to −2400 µm) and caudal (+1800 to +3600 µm) to the lesion (Control n = 18; epoD n = 21; epoB n = 12; rehabilitation n = 9; epoD + rehabilitation n = 12; epoB + rehabilitation n = 10). ( F ) Schematic representation of the region in which IHC was performed for the colocalization of fibronectin to RECA1 and CD13. Created with BioRender.com. ( G ) Fluorescent images of spinal cord sections 3600 µm rostral from the lesion epicentre labelled for fibronectin and RECA-1 or CD13 (scale bar = 50 µm; 10 µm in zoomed panels). Plotted data are the mean ± SEM; each data point represents the mean per animal. * P < 0.05, ** P < 0.01, *** P < 0.001 by two-way ANOVA and Bonferroni post hoc.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: Staining

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: Epothilones but not rehabilitation promotes serotonergic and lumbar VGluT1 plasticity . ( A ) Representative images of sections 1800–3000 µm caudal to the lesion immunolabelled for 5-HT expression (pixel values inverted; scale bar = 200 µm). ( B ) Scheme depicting the ventral horn region in which images were thresholded to positive staining area and total 5-HT + immunolabelling within the ventral horns were measured. Created with BioRender.com. ( C ) Quantification of 5-HT staining area in the ventral horns in the given treatment groups (control n = 15; epoD n = 18; epoB n = 11; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 10). ( D ) Representative fluorescent images of spinal cord sections immunolabelled for VGluT1 (grey value intensity gradient; scale bar = 500 µm) in the given treatment groups in coronal sections from lumbar spinal cord (L1–L4). ( E ) Scheme depicting that the quantification for VGluT1 was segregated into Laminae I–II, III–V and V–IX. Created with BioRender.com. ( F – H ) Quantification of VGluT1 immunolabelling per area in the various laminae in the given treatment groups (control n = 9; epoD n = 9; epoB n = 12; rehabilitation n = 11; epoD + rehabilitation n = 10; epoB + rehabilitation n = 10). Plotted data are the mean ±SEM; each data point represents the mean per animal. * P < 0.05, ** P < 0.01, *** P < 0.001 by repeated-measures two-way ANOVA Dunnett’s and Bonferroni post hoc.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: Expressing, Staining

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: Rehabilitation and a combination of rehabilitation with epothilone B promote recovery in open field and ladder tasks . ( A and B ) The actual force and displacement of each impaction were plotted by one-way ANOVA, Bonferroni post hoc . ( C and D ) BBB and horizontal error ladder scores measured throughout the study. Plotted data are the mean ±SEM; each data point represents the mean per animal. * P < 0.05, ** P < 0.01 by repeated-measures two-way ANOVA Dunnett’s and Bonferroni post hoc (light blue asterisks compare control with epoB = rehab; grey asterisks compare control with rehab). Control n = 17; epoD n = 20; epoB n = 11; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 11.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques:

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: Epothilone B improves aspects of hindlimb kinematics . ( A ) Schematic of the five measured marker positions on the hindlimbs. For each video frame, these marker positions were used to calculate joint angles as depicted here. Created with BioRender.com. ( B ) An example of a complete walking trial for a right hind leg (uninjured condition). Marker colours correspond to those in ( A ). Complete step cycles have been defined as the movement from a lift-off event to the next. ( C ) Time course of all joint angles [see ( A )] averaged over all steps for each individual and then averaged for all treatment groups separately (only left body side shown), normalized to their respective step cycles (see for averages of all individuals and both body sides). ( D ) Absolute amplitudes (difference between the maximum and minimum value during a step cycle) of all individual joint angles in all conditions (* P < 0.05, ** P < 0.01, *** P < 0.001, Wilcoxon rank-sum test; black asterisks compare with control whereas brown asterisks compare with uninjured). ( E ) Average height of the IC marker above the surface of the walkway in all individual trials. Height was normalized to the height in the baseline condition. Boxplots indicate median, 25th, and 75th percentile (box) and 5th and 95th percentile (whiskers). Uninjured = 31; control n = 9; epoD n = 10; epoB n = 11; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 11.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: Marker

Journal: Brain Communications

Article Title: Rehabilitation enhances epothilone-induced locomotor recovery after spinal cord injury

doi: 10.1093/braincomms/fcad005

Figure Lengend Snippet: The combination of rehabilitation with epothilone B restores parameters of gait through complementary locomotor mechanisms . ( A ) An example image of the Catwalk XT gait analysis system recording paw prints during a run. ( B ) Heatmap hierarchical clustering of the PCA data set for correlation of parameters between treatment groups. ( C ) Three-dimensional depiction of the projection of all individual trials (data points represents means per animal) to the first three PCs and scores for PCs 1 and 2 (see also ). Boxplots indicate the median, 25th and 75th percentile (box) and 5th and 95th percentile (whiskers). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by repeated-measures two-way ANOVA Dunnett’s post hoc . Uninjured = 31; control n = 28; epoD n = 28; epoB n = 11; rehabilitation n = 10; epoD + rehabilitation n = 12; epoB + rehabilitation n = 10.

Article Snippet: Following the dosing scheme of our previous studies, animals were randomly allocated to treatment groups and received blinded s.c. injections of either epoD (1.5 mg/kg; Abcam, ab143616), epoB (0.75 mg/kg; Selleck Chemicals S1364) or vehicle (50% DMSO) on Days 1 and 15 post-injury.

Techniques: