dylight 649 conjugated lectin  (Vector Laboratories)

Journal: Cardiovascular Research

Article Title: Age-dependent remodelling of arterial chemoafferent innervation in hypertension

doi: 10.1093/cvr/cvaf207

Figure Lengend Snippet: Carotid body hypertrophy in SHR precedes the onset of hypertension. ( A ) Transverse CB area plotted for each serial section along the rostral–caudal axis. ( B ) CB size in the sagittal axis is measured as the number of 20 µm sections spanning the CB. Coloured dots indicate observed values for each animal. Black dots represent the mean value across strains within each age group. Boxplots represent strain within each age group. ( C ) Data from panel A replotted so that the largest CB transverse area in each animal is set as the centre (0) along the rostral–caudal axis. ( D ) Cumulative CB area where the largest transverse area in each animal is assigned as the centre (0) along the rostral–caudal axis. ( E ) Representative image of CB size and shape in the transverse plane. Example selected from young SHR and Wistar rats, showing the section with the widest (0) area closest to the group average. TH—tyrosine hydroxylase (green). NfM—neurofilament-M (magenta). TL/LEL—tomato ( Lycopersicon esculentum ) lectin (white). ( F ) A representative 3D render of CB size and shape in aged SHR and Wistar rats. Area outlines are centred on the xy coordinate plane and manually adjusted for radial alignment. n = 1. For A and C , lines represent smoothed trends calculated using LOESS estimation. In A , C , and D , shaded areas represent a 95% confidence interval. Sample size—Young: W, n = 7, N = 150; SHR, n = 8, N = 260; Aged: W, n = 8, N = 272; SHR, n = 10, N = 432; Old: W, n = 6, N = 145; SHR, n = 8, N = 396 ( n —biological replicates, N —observation). Hypothesis testing was performed on data summarized in A and B using a generalized additive model (GAM) with log-transformed data fitted to a Gaussian distribution. For C , a generalized linear model (GLM) was fitted to a Poisson distribution. P -values were derived from a post-hoc estimated marginal means test for specified comparison groups ( n = 9) using false discovery rate (FDR) multiple comparison correction.

Article Snippet: Sections were rinsed 3 × 10 min in PBS and incubated in the secondary antisera solution containing secondary antibodies, Lycopersicon esculentum (tomato) lectin conjugated to DyLight®-647 (1:250; DL-1178-1; Vector Laboratories) and DAPI (1:10 000; D9542; Sigma-Aldrich) for 2 h at room temperature.

Techniques: Transformation Assay, Derivative Assay, Comparison

Journal: Cardiovascular Research

Article Title: Age-dependent remodelling of arterial chemoafferent innervation in hypertension

doi: 10.1093/cvr/cvaf207

Figure Lengend Snippet: Increased carotid body vascularization in SHR. ( A ) Representative image of the capillary network perfusing the CB. The yellow line outlines the CB area. TL/LEL—tomato ( Lycopersicon esculentum ) lectin (white). ( B , C ) Tomato lectin-positive area in the CB plotted for each serial section along the rostral–caudal axis ( B ) and relative to the largest transverse area in each animal set as the centre (0) along the rostral–caudal axis ( C ). ( D , E ) Proportion of CB area labelled by TL/LEL summarized for each group ( D ) and plotted relative to the largest transverse area in each animal set as the centre (0) along the rostral–caudal axis ( E ). In D , observed values (coloured dots) for each biological replicate (animal) are aligned vertically. Black dots represent the mean value across strains within each age group. Boxplots represent strain within each age group. For B , C , and E , lines represent smoothed trends calculated using LOESS estimation, and shaded areas represent the 95% confidence interval. Sample size—Young: W, n = 7, N = 142; SHR, n = 8, N = 238; Aged: W, n = 8, N = 271; SHR, n = 10, N = 428; Old: W, n = 6, N = 144; SHR, n = 8, N = 396 ( n —biological replicates, N —observation). Hypothesis testing was performed using a generalized additive model (GAM) with log-transformed data fitted to a Gaussian distribution. P -values were derived from a post-hoc estimated marginal means test for specified comparison groups ( n = 9) using false discovery rate (FDR) multiple comparison correction.

Article Snippet: Sections were rinsed 3 × 10 min in PBS and incubated in the secondary antisera solution containing secondary antibodies, Lycopersicon esculentum (tomato) lectin conjugated to DyLight®-647 (1:250; DL-1178-1; Vector Laboratories) and DAPI (1:10 000; D9542; Sigma-Aldrich) for 2 h at room temperature.

Techniques: Transformation Assay, Derivative Assay, Comparison

Journal: Cardiovascular Research

Article Title: Age-dependent remodelling of arterial chemoafferent innervation in hypertension

doi: 10.1093/cvr/cvaf207

Figure Lengend Snippet: Age-dependent remodelling of chemoafferent innervation of the carotid body. ( A ) Two types of nerve fibres (NF + , TH + and NF + , TH − ) innervating the CB. Arrowheads mark NF + , TH + fibres. ( B ) Sympathetic motor terminals (NF − , TH + ) in close association with CB vasculature (arrowheads). Double arrowheads mark NF + , TH − fibres. Asterisk mark TH + chemosensory cells. TH—tyrosine hydroxylase (green). NfM—neurofilament-M (magenta). TL/LEL—tomato ( Lycopersicon esculentum ) lectin (white). Scale bar—10 μm. Scale bar (insets)—1 μm. C—Area of NF + , TH − fibres and ( D ) NF + , TH + fibres in the CB. ( E ) Proportion of NF-immunoreactive area corresponding to NF + , TH − /NF + , TH + fibres. ( F , G ) Proportion of the CB area occupied by NF + , TH − ( F ), and motor ( G ) fibres. In C – G , observed values (coloured dots) for each biological replicate (animal) are aligned vertically. In C , D , F , and G , black dots represent the mean value across strains within each age group. Boxplots represent strain within each age group. In E , each section is represented by two values comprising a total of 100%: (i) percentage of NF + area co-localizing with TH (NF + , TH + ) and (ii) remaining NF + area (NF + , TH − ). Sample size—Young: W, n = 7, N = 149; SHR, n = 8, N = 237; Aged: W, n = 8, N = 269; SHR, n = 10, N = 430; Old: W, n = 6, N = 142; SHR, n = 8, N = 396 ( n —biological replicates, N —observation). Hypothesis testing was performed using a generalized additive model (GAM) with log-transformed data fitted to a Gaussian distribution. P -values were derived from a post-hoc estimated marginal means test for specified comparison groups ( n = 9) using false discovery rate (FDR) multiple comparison correction.

Article Snippet: Sections were rinsed 3 × 10 min in PBS and incubated in the secondary antisera solution containing secondary antibodies, Lycopersicon esculentum (tomato) lectin conjugated to DyLight®-647 (1:250; DL-1178-1; Vector Laboratories) and DAPI (1:10 000; D9542; Sigma-Aldrich) for 2 h at room temperature.

Techniques: Transformation Assay, Derivative Assay, Comparison